Table of Content

18 August 2022, Volume 26 Issue 23 Previous Issue    Next Issue

For Selected:

Download Citations EndNote Reference Manager ProCite BibTeX RefWorks

Toggle Thumbnails

Select

Interleukin-1 alpha induces osteoclast activation and bone loss Yang Ruijuan, Li Yangyang, Cai Ruiyan, Liu Huibin, Guo Chun 2022, 26 (23):  3691-3699.  doi: 10.12307/2022.669 Abstract ( 490 )   PDF (16094KB) ( 32 )   Save BACKGROUND: Interleukin-1 is an important pro-inflammatory cytokine that has been documented in the regulation of bone inflammation and bone remodeling. A previous study has demonstrated that interleukin-1α can induce apoptosis while inhibiting osteoblast differentiation in MC3T3-E1 cells.  OBJECTIVE: To investigate the role and mechanism of interleukin-1α on osteoclast activation and bone loss in mice. METHODS: (1) Cell test: RAW264.7 cells were either treated with interleukin-1α alone or with receptor activator of nuclear factor-κB ligand (RANKL) for 1 and 4 days. Cell viability was tested by cell counting kit-8 assay. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The mRNA and protein levels of osteoclast-specific genes and genes related to nuclear factor-κB pathway and Wnt/β-catenin pathway were tested by real-time fluorescence quantitative PCR, immunofluorescence staining or western blot. Bone marrow-derived macrophages were either treated with interleukin-1α alone or with RANKL and macrophage colony-stimulating factor for 7 days. The number of multinuclear osteoclasts was detected by tartrate resistant acid phosphatase assay. The protein levels of osteoclast-specific genes were tested by western blot. (2) Animal test: Twenty-four male C57BL/6J mice (6-8 weeks old) were assigned into two groups at random: control group  and test group. Mice were subsequently treated with interleukin-1α solution or PBS by intraperitoneal injection twice a week for 5 weeks. Bone tissues from the femurs were performed with micro-computed tomography analysis and hematoxylin-eosin staining, tartrate resistant acid phosphatase, and immunofluorescence analysis.  RESULTS AND CONCLUSION: Cell test: Interleukin-1α alone significantly increased RAW264.7 cell proliferation, but stimulated cell differentiation into osteoclasts in combination with RANKL (P < 0.05). Interleukin-1α significantly increased the expression of osteoclast-related markers and the number of tartrate resistant acid phosphatase-positive multinuclear cells in RAW264.7 cells and bone marrow-derived macrophages in the existence of RANKL or RANKL+macrophage colony-stimulating factor (both P < 0.05). Interleukin-1α was found to significantly enhance the nuclear factor-κB and Wnt/beta-catenin signaling in RAW264.7 cells (P < 0.05). Blocking of nuclear factor-κB or Wnt3 signaling not only reversed the activation of nuclear factor-κB and Wnt3 signaling but also weakened the enhanced expression of osteoclast-specific genes induced by interleukin-1α in RAW264.7 cells (P < 0.05). Animal test: interleukin-1α induced bone loss in mice while also upregulating the expression of osteoclast-specific markers, RANK, TRAF6 and p65, and Wnt3 in vivo (P < 0.05). The findings indicate that interleukin-1α can induce osteoclast activation and bone loss by promoting the nuclear factor-κB and Wnt signaling pathways.  Figures and Tables | References | Related Articles | Metrics

Select

Differences of trabecular microstructure and bone metabolism in two types of kidney-deficiency ovariectomized rats He Xingpeng, Zheng Liqin, Li Pengfei, Yue Guiyang, Li Zhihong, Wu Minhui, Lin Ziling 2022, 26 (23):  3768-3772.  doi: 10.12307/2022.681 Abstract ( 401 )   PDF (2379KB) ( 73 )   Save BACKGROUND: At present, ovariectomized rats are a mature and reliable animal model of osteoporosis, and establishment and observational evaluation of rat models of two types of kidney deficiency have been relatively detailed. However, further exploration is needed on the rat models of osteoporosis combined with syndromes of kidney-yang deficiency and kidney-yin deficiency. The sympathetic-parasympathetic nerve function test is used as a specific index to judge the syndrome of yin-deficiency and yang-deficiency, but this indicator is relatively insufficient for evaluating and analyzing the skeletal system of the two types of kidney-deficiency syndromes. “Integration of disease and syndrome” requires more experimental evidence and observational indicators. OBJECTIVE: To compare trabecular microstructure parameters and serum bone metabolism indicators between osteoporosis rat models of kidney-yang deficiency and kidney-yin deficiency. METHODS: The ovaries of 20 female Spraque-Dawley rats were surgically removed to make postmenopausal osteoporotic rat models. Ten weeks after operation, all model rats were randomly divided into two groups (n=10 per group): kidney-yang deficiency group and kidney-yin deficiency group. Gluteal intramuscular injection of hydrocortisone and intragastric administration of thyroid hormone tablets were given to the kidney-yang deficiency group and the kidney-yin deficiency group, respectively, once a day for 14 consecutive days. After modeling, micro-CT was used to detect trabecular microstructure parameters in the region of interest of the left femur, and ELISA was used to detect serum bone formation and bone resorption indicators. RESULTS AND CONCLUSION: Two types of kidney-deficiency osteoporosis rat models were successfully established. Compared with the kidney-yang deficiency group, bone mineral density, bone volume/total volume, and trabecular number were significantly increased, trabecular thickness decreased, and serum bone formation and bone resorption indicators significantly increased in the kidney-yin deficiency group. Therefore, bone metabolism was faster in the kidney-yin deficiency group than the kidney-yang deficiency group. All these findings indicate significant differences in trabecular microstructure parameters and bone metabolism indicators between two kidney-yang and kidney-yin deficiency groups, and the mechanisms need to be further studied. Figures and Tables | References | Related Articles | Metrics

Select

Isolation, culture and identification of rabbit auricular chondrocytes Zhong Ziling, Qu Shenhong, Han Xing, Wu Di 2022, 26 (23):  3633-3637.  doi: 10.12307/2022.660 Abstract ( 399 )   PDF (2590KB) ( 47 )   Save BACKGROUND: Auricle deformity remodeling based on three-dimensional printing scaffold materials has become a focus in recent years, and whether chondrocytes can grow normally on a scaffold is the key issue.   OBJECTIVE: To investigate the primary culture methods of rabbit auricular chondrocytes, thereby laying a solid foundation for the application of chondrocytes combined with tissue engineering.  METHODS: Rabbit auricular chondrocytes were obtained by trypsin and type II collagenase sequential digestion method. The chondrocytes were cultured and subcultured in vitro. The chondrocytes were identified by morphological observation, growth curve drawing, safranin O-fast green staining, toluidine blue staining, Victoria blue staining, and glycosaminoglycan content determination.  RESULTS AND CONCLUSION: The normal morphology of auricular chondrocytes was fusiform and polygonal under inverted phase contrast microscope. After six generations of subculture, the cells became obviously long fusiform, and the proliferation rate slowed down or even stopped. The growth curve showed that the chondrocytes from the 2nd to 5th generations could proliferate in a straight line. Results of safranin O-fast green staining showed clear nucleus and cytoplasm structures of rabbit auricular chondrocytes, with good morphological characteristics. Results of toluidine blue staining indicated that the secretion of cartilage matrix was less at the initial stage of culture and increased gradually with the increase of cell fusion. Results of Victoria blue staining showed the formation of collagen fibers in chondrocytes, suggesting the normal function of chondrocytes. Results of glycosaminoglycans content determination showed the specific secretion of glycosaminoglycans in chondrocytes. Overall, the isolation and identification system of rabbit auricular chondrocytes in primary culture was successfully established, and the auricular chondrocytes with biological activity were obtained, which could be further used in tissue engineering research.  Figures and Tables | References | Related Articles | Metrics

Select

Screening of drugs for inhibiting growth plate chondrocyte apoptosis and the effect of resveratrol on delaying epiphyseal closure in rats Xie Yan, Li Wuyin, Xing Weipeng, Pei Yuanyuan, Wang Na 2022, 26 (23):  3644-3649.  doi: 10.12307/2022.662 Abstract ( 543 )   PDF (2037KB) ( 47 )   Save BACKGROUND: Epiphyseal closure indicates the long bones stop growing, which is caused by the apoptosis of growth plate chondrocytes. It has been reported that aromatase inhibitors can inhibit the production of estrogen, and the cessation of bone growth is caused by epiphyseal closure. Therefore, it has been inferred that aromatase inhibitors can inhibit growth plate chondrocyte apoptosis and delay epiphyseal closure.   OBJECTIVE: To compare the effects of several aromatase inhibitors (resveratrol, biochanin A, and flax lignans) on inhibiting the apoptosis of rat growth plate chondrocyte and to verify the effect of resveratrol on delaying epiphyseal closure and promoting the growth of long bones in rats. METHODS:  Rat tibial and femoral growth plate cartilage samples were dissected, and growth plate chondrocytes were isolated and cultured in vitro by two-step enzyme digestion. Chondrocytes were morphologically observed and identified by inverted phase contrast microscope and type II collagen immunofluorescence experiments. Then interleukin 1β was used to induce apoptosis of growth plate chondrocytes. Cells were cultured with culture medium containing 10, 20, 40, and 60 μmol/L resveratrol, with culture medium containing 5, 10, 20, 30 μmol/L biochanin, or with culture medium containing 1, 10, 20, 40 μmol/L flax lignans to compare the effects of different concentrations of drugs on the apoptosis of growth plate chondrocytes. Resveratrol which had the best effect was administered to rats by gavage to investigate its effect on the epiphyseal closure of rat tibial plateau and the growth of long bones.   RESULTS AND CONCLUSION: Appropriate concentrations of resveratrol, biochanin A, and flax lignans could inhibit the apoptosis of growth plate chondrocytes to varying degrees. The best drug concentrations for inhibiting cell apoptosis were 40, 10, and 20 μmol/L in sequence, and 40 μmol/L resveratrol had the best effect on inhibiting the apoptosis of growth plate chondrocytes. Administration of 40 mg/kg resveratrol by gavage could delay epiphyseal closure of tibial plateau and promote the growth of long bones in rats. Therefore, resveratrol can promote bone growth during developmental period by prolonging the time of bone growth. Figures and Tables | References | Related Articles | Metrics

Select

Protective effect of eucommia alcohol extract on articular cartilage of osteoarthritis model rats Yang Demeng, Wang Changgeng, Hu Xinyuan, Ao Fei 2022, 26 (23):  3756-3761.  doi: 10.12307/2022.679 Abstract ( 525 )   PDF (2739KB) ( 97 )   Save BACKGROUND: At present, studies have found that eucommia water extract can significant relieve osteoarthritis, but there is no report about the therapeutic effect of eucommia alcohol extract on osteoarthritis.  OBJECTIVE: To investigate the protective effect of eucommia alcohol extract on osteoarthritis rats, and the mechanism of action of the janus kinase 1 (JAK1)/signal transducer and activator of transcription 3 (STAT3)/suppressor of cytokine signaling 3 (SOCS3) pathway.  METHODS: Sprague-Dawley rats were randomly divided into four groups (n=10 per group): a sham operation group, a model group, a low-dose eucommia alcohol extract group, and a high-dose eucommia alcohol extract group. Transection of the medial meniscotibial ligament of the right knee joint was conducted to induce an osteoarthritis rat model in all the groups except for the sham operation group. Rats in the low-dose and high-dose eucommia alcohol extract groups were given 50 and 200 mg/kg eucommia alcohol extract by intragastric administration for 8 weeks. Gait behaviors were tested, and paw withdrawal mechanical threshold and paw withdrawal thermal latency were determined. The changes of rat joint space and cartilage surface calcification were evaluated with X-ray imaging. The levels of inflammatory factors interleukin 6, interleukin 1β, and tumor necrosis factor α in rat serum were detected. Pathological changes of rat articular cartilage were observed using Safranin-O staining and hematoxylin-eosin staining, and were evaluated by Osteoarthritis Research Society International scoring system. Western blot was used to detect the protein expression of JAK1/STAT3/SOCS3 pathway in rat articular cartilage tissue. RESULTS AND CONCLUSION: The rat cartilage surface was smooth in the sham operation group, but was severely damaged in the model group. Moreover, the cartilage surface density was increased, and the joint space became narrowed in the model group. Low- and high-dose eucommia alcohol extracts significantly relieved cartilage injury, and reduced surface calcification. Compared with the sham operation group, the Osteoarthritis Research Society International score, paw withdrawal mechanical threshold value, levels of interleukin 6, interleukin 1β, and tumor necrosis factor α, and protein expression of matrix metalloproteinase 13, JAK1, and p-STAT3 were significantly increased (P < 0.05), while the gait behavior score, paw withdrawal thermal latency and the protein expression of SOCS3 were significantly reduced in the model group (P < 0.05). Compared with the model group, the Osteoarthritis Research Society International score, paw withdrawal mechanical threshold value, levels of interleukin 6, interleukin 1β, tumor necrosis factor α, and protein expression of matrix metalloproteinase 13, JAK, and p-STAT3 were significantly reduced (P < 0.05), while the gait behavior score, paw withdrawal thermal latency and the protein expression of suppressor of cytokine signaling 3 were significantly increased in the low- and high-dose eucommia alcohol extract groups (P < 0.05). Therefore, eucommia alcohol extract can inhibit the JAK1/STAT3 pathway, promote the protein expression of SOCS3 and reduce inflammation, thereby protecting the articular cartilage of osteoarthritis rats.  Figures and Tables | References | Related Articles | Metrics

Select

Mechanisms of Huangqi Guizhi Wuwu Decoction in the treatment of cervical spondylosis and anxiety disorder based on the principle of “treating different diseases with the same method”: a network pharmacology analysis Deng Bowen, Li Xiaoye, Jiang Shengyuan, Liu Gan, Zhao Yi, Zhang Houjun, He Feng, Mu Xiaohong 2022, 26 (23):  3650-3656.  doi: 10.12307/2022.663 Abstract ( 567 )   PDF (9830KB) ( 36 )   Save BACKGROUND: Huangqi Guizhi Wuwu Decoction is widely used in the treatment of cervical spondylosis and some affective disorders, but its specific mechanism is still unclear. OBJECTIVE: Through the methods of network pharmacology and molecular docking, to preliminarily discuss the common mechanism of Huangqi Guizhi Wuwu Decoction in the treatment of cervical spondylosis and anxiety disorder according to the treating principle of “treating different diseases with the same method.” METHODS: We searched the pharmacology database of traditional Chinese medicine system from the inception to September 1, 2021, retrieving the effective active components and corresponding targets of five drugs in Huangqi Guizhi Wuwu Decoction. Genecards, OMIM, PharmGKB, TTD and drugbank databases were searched using the keywords of “cervical spondylosis” and “anxiety disorder” from their inception to September 1, 2021, and the disease targets of cervical spondylosis and anxiety disorder were mined. The key targets of Huangqi Guizhi Wuwu Decoction in the treatment of cervical spondylosis and anxiety disorder between components and disease targets were screened using R language to construct the active ingredient-target-disease and protein-protein interaction network through the Cytoscape 3.8.2 and STRING, as well as screen the common targets of network core. The intersection targets were obtained by GO enrichment analysis and KEGG pathway enrichment by R language. The Vina and PyMOL software were to establish molecular docking between common targets and main active ingredients in the protein-protein interaction network. RESULTS AND CONCLUSION: We screened 73 active components of Huangqi Guizhi Wuwu Decoction in total, including quercetin, kaempferol β-Carotene. There were 19 targets associated with the disease, and the key targets included interleukin-1β, TP53, hypoxia-inducible factor-1α, mitogen activated protein kinase 1, etc. Molecular docking results showed that kaempferol could target and regulate interleukin-1β, TP53, and hypoxia-inducible factor-1α. GO preliminary analysis showed that the mechanism of Huangqi Guizhi Wuwu Decoction in the treatment of cervical spondylosis and anxiety mainly involved biological processes including reactive oxygen species metabolism, regulation of inflammatory response, oxidative stress response and cell apoptosis. KEGG results showed that according to the principle of “treating different diseases with the same method,” Huangqi Guizhi Wuwu Decoction played the therapeutic role in cervical spondylosis and anxiety disorder by regulating 126 common pathways such as interleukin-17 signal pathway, tumor necrosis factor signal pathway and mitogen activated protein kinase signal pathway. All these findings preliminarily reveal that Huangqi Guizhi Wuwu Decoction follows the principle of “treating different diseases with the same method” in the treatment of cervical spondylosis and anxiety disorder, which may play a role in regulating neuroimmune inflammation, antioxidant stress, cell apoptosis and other related biological processes through active components such as quercetin and kaempferol, so as to provide new ideas for clinical medication and theoretical basis for basic experiments. Figures and Tables | References | Related Articles | Metrics

Select

An appropriate dose of intra-articular medical ozone injection in a rat model of temporomandibular joint osteoarthritis Zhang Wanxia, Nijiati·Nuermuhanmode, Maisituremu·Heilili, Liu Jianglong, Liu Di, Maimaitituxun·Tuerdi 2022, 26 (23):  3700-3705.  doi: 10.12307/2022.670 Abstract ( 522 )   PDF (5100KB) ( 81 )   Save BACKGROUND: Medical ozone is a mixture of ozone and oxygen that, in contact with body fluids, rapidly produces reactive oxygen species and activates the body’s antioxidant system. Clinically, the intra-articular injection of medical ozone can effectively alleviate the progress of osteoarthritis. OBJECTIVE: To study the therapeutic effect of intra-articular injection of medical ozone in a rat model of temporomandibular joint osteoarthritis. METHODS: The rat model of temporomandibular joint osteoarthritis was established through intra-articular injection of sodium iodoacetate. Medical ozone with mass concentrations of 20, 30 and 40 mg/L was then injected into the joint cavity of the experimental rats once a week for 3 weeks. Model group and control group were injected with the equal amount of saline. The structural changes of the temporomandibular joint were evaluated by cone beam CT examination 1 week after the last injection. The tumor necrosis factor α level in the condylar cartilage and plasma was measured by ELISA, and the tissue samples of the temporomandibular joint on the other side were stained with hematoxylin-eosin and safranin O-fast green, and histopathological changes of the temporomandibular joint were evaluated by the modified Mankin’s score. RESULTS AND CONCLUSION: Cone beam CT results indicated that the temporomandibular joint and condylar process of control rats had regular shape and smooth surface, while condylar process deformation and surface defects of rats in the medical ozone 20 and 30 mg/L groups were smaller than those in the model and medical ozone 40 mg/L groups. ELISA results revealed that the expression levels of tumor necrosis factor α in the cartilage and plasma of the rat condylar process were significantly increased in the model group and medical ozone 20, 30, 40 mg/L groups compared with the control group (P < 0.05). Compared with the model group and medical ozone 40 mg/L group, the expression level of tumor necrosis factor α in the cartilage and plasma were significantly decreased in the medical ozone 20, 30 mg/L groups (P < 0.05). Histological observation revealed that the modified Mankin’s score in the medical ozone 20, 30, 40 mg/L groups and the model group were significantly higher than that in the control group (P < 0.05); the modified Mankin’s score in the medical ozone 20, 30 mg/L groups was significantly lower than that in the model group and the medical ozone 40 mg/L group (P < 0.05). To conclude, the intra-articular injection of 20 and 30 mg/L medical ozone can significantly delay the progression of osteoarthritis in the temporomandibular joint of rats, but when the mass concentration of ozone is increased to 40 mg/L, it will aggravate osteoarthritis in the temporomandibular joint of rats. Figures and Tables | References | Related Articles | Metrics

Select

Interleukin-27 is associated with the pathogenesis of lumbar disc herniation Xue Huawei, Zhu Min, Li Yuqian 2022, 26 (23):  3750-3755.  doi: 10.12307/2022.678 Abstract ( 485 )   PDF (1483KB) ( 60 )   Save BACKGROUND: Inflammation is an important mechanism underlying lumbar disc herniation. Th17 cells play a key role in immune activation. Interleukin-27 can inhibit Th17 cell differentiation by down-regulating the expression of interleukin-6 and transforming growth factor-β, and plays an important role in a variety of autoimmune diseases. However, little is known about the relationship between interleukin-27 and lumbar disc herniation. OBJECTIVE: To investigate the levels of interleukin-27 in patients with lumbar disc herniation, and to explore their association with lumbar intervertebral disc herniation. METHODS: This study enrolled 36 patients with lumbar disc herniation, 18 healthy controls, and 8 patients undergoing emergency surgery for thoracolumbar burst fracture. Patients with lumbar disc herniation received posterior fenestration discectomy, posterior lumbar interbody fusion (PLIF) or transforaminal lumbar interbody fusion. The pain intensity of the patients was scored using a visual analogue scale (VAS) before operation. Serum interleukin-27 and interleukin-17 levels in peripheral blood were determined by enzyme linked immunosorbent assay (ELISA). Samples of nucleus pulposus from patients with lumbar disc herniation and those with thoracolumbar burst fractures (considered as normal nucleus pulposus samples) were taken for detection of interleukin-27 and interleukin-17 mRNA and protein using quantitative real-time PCR and western blot assay, respectively. RESULTS AND CONCLUSION: Patients with lumbar disc herniation had significantly higher serum interleukin-27 levels than healthy controls (P < 0.001). Interleukin-17 levels were also significantly higher in the patients with lumbar disc herniation. The protein and mRNA expression levels of interleukin-27 and interleukin-17 in the nucleus pulposus were significantly higher in patients with lumbar disc herniation than patients with thoracolumbar burst fractures (P < 0.001).  The expression of interleukin-27 was significantly higher in patients with mild pain than those with severe pain (P < 0.01). To conclude, inflammation is responsible for the pain experienced by patients with lumbar disc herniation, and interleukin-27 is involved in the pathogenesis of lumbar disc herniation. Figures and Tables | References | Related Articles | Metrics

Select

Determination and characteristics of skeletal muscle output power in different strength training methods under optimal power load Liang Meifu, Guo Wenxia, Zhao Ningning, Pan Lei 2022, 26 (23):  3638-3643.  doi: 10.12307/2022.661 Abstract ( 610 )   PDF (3546KB) ( 68 )   Save BACKGROUND:  Strength training under the optimal power load can maximize skeletal muscle output power. The increase of skeletal muscle output power can improve athletes’ sports performance and public health level, which is favored by scholars and physical coaches at home and abroad. OBJECTIVE: To determine the optimal power load of different strength training methods and analyze the output power characteristics. METHODS: Twenty-seven male college athletes from Beijing Sport University were recruited for testing 1 repetition maximum (1RM) of half squat and bench press throw. Nine axis sensors were used to test the output power characteristics of subjects under the maximum power loads of 10%, 30%, 50%, 70%, and 90%. RESULTS AND CONCLUSION: The optimal power load of half squat was 70% 1RM, and its mean output power and maximum output power were significantly higher than 10% 1RM, 30% 1RM, and 50% 1RM (P < 0.01). The output power of half squat was highly correlated with the maximum velocity (r=0.84-0.87, P < 0.01), moderately correlated with the mean velocity and mean force (r=0.50-0.68, P < 0.01), and low correlated with 1RM (r=0.40, P < 0.05). The optimal power load of bench press throw  was 70% 1RM, and its mean output power was significantly higher than 10% 1RM (P < 0.01), 30% 1RM (P < 0.05), and 90% 1RM (P < 0.01). The output power of bench press was highly correlated with the mean velocity, maximum velocity and maximum power (r=0.70-0.84, P < 0.01), moderately correlated with the mean force and 1RM (r=0.57-0.70, P < 0.01), and low correlated with body mass and training years (r=0.40-0.46, P < 0.05). To conclude, the mean value of the optimal power load of half squat and bench press throw  of college athletes is 70% 1RM. Due to the difference of individual physiology and training, the accurate optimal power load of skeletal muscle still needs further measurement. To improve the maximum output power of skeletal muscle, developing skeletal muscle contraction velocity is prior to developing skeletal muscle contraction force in college athletes. Figures and Tables | References | Related Articles | Metrics

Select

Effect of p38 inhibitor on inflammatory factors in the patellar tendon and patella-patellar tendon junction under short-term exercise Qi Yanan, Chen Yiyan, Zhang Ning, Wang Lin 2022, 26 (23):  3744-3749.  doi: 10.12307/2022.677 Abstract ( 452 )   PDF (2907KB) ( 80 )   Save BACKGROUND: p38 inhibitors have achieved good results in the intervention and treatment of related diseases. However, there are still many questions about the effect of p38 inhibitors on the inflammatory response in the patellar tendon and the patella-patellar tendon junction under short-term high-intensity exercise. OBJECTIVE: On the basis of a quantitative jumping animal model successfully established in the laboratory, to investigate the effect of p38 inhibitors on the expression of main inflammatory factors, including interleukin-1β, interleukin-6 and transforming growth factor-β1 in the patellar tendon and patella-patellar tendon junction under short-term high-intensity exercise. METHODS: Thirty-four 18-week-old New Zealand white rabbits were randomly divided into control group (n=4); jump 1, 3, 5 days groups (n=5 per group); jump+p38 inhibition 1, 3, 5 days groups (n=5 per group). Each jump group was subjected to electrical stimulation-induced jump training (jumping forward and upward), with a jump height of 10 cm defined as a qualified jump. Each training sessions included 150 qualified jumps. SB203580 (0.5 mg/kg) was intraperitoneally injected in each jump+p38 inhibition group after each training. The control group did not perform jump training, and the experimental group received electrical stimulation jump training according to the training program. The pre-adaptation process and feeding time were the same. Except for the control group, the rabbits from the other groups were sacrificed after training, and immunohistochemical staining was performed using the SABC method, and the density of the positive cells of each inflammatory factor was quantitatively calculated by Metamorph graphics processing software. RESULTS AND CONCLUSION: Immunohistochemistry results indicated inconsistent changes in the expression of interleukin-1β, interleukin-6 and transforming growth factor-β1 in the patellar tendon and patella-patellar tendon junction under short-term high-intensity exercise. Compared with the jump groups, the expressions of interleukin-1β, interleukin-6 and transforming growth factor-β1 in the patellar tendon and patella-patellar tendon junction were not significantly different in each jump+p38 inhibitor group (P > 0.05). These findings reveal that the p38 inhibitor may not affect the inflammatory response of the patella tendon and patella-patellar tendon junction in the acute phase under short-term high-intensity exercise. Figures and Tables | References | Related Articles | Metrics

Select

Asiaticoside protects the kidney from ischemia-reperfusion injury in a rat model by activating nuclear factor E2-related factor 2/hemeoxygenase-1 pathway Zhu Shiyu, Hu Yan, Wang Suogang, Lu Peng, Wang Di, Zhai Qiongyao, Wang Guangce 2022, 26 (23):  3609-3614.  doi: 10.12307/2022.657 Abstract ( 386 )   PDF (4883KB) ( 68 )   Save BACKGROUND: Prevention or reduction of kidney ischemia-reperfusion injury can improve the livability of kidney transplant. Therefore, it is necessary to develop safer, cheaper and more effective drugs to prevent or treat kidney ischemia-reperfusion injury.   OBJECTIVE: To investigate the protective effect of asiaticoside on kideny ischemia-reperfusion injury in a rat model and its relevant mechanism.   METHODS: A total of 40 Sprague-Dawley rats were randomly divided into four groups: sham surgery group, model group, asiaticoside group, and asiaticoside+brusatol group, with 10 rats in each group. The kidney pedicles were clamped for 45 minutes in the latter three groups to establish the rat kidney ischemia-reperfusion injury model. The rats in the asiaticoside and asiaticoside+brusatol groups were given asiaticoside suspension by gavage for 4 weeks in advance. Before modeling, the asiaticoside+brusatol group was given brusatol, a nuclear factor E2-related factor 2-specific inhibitor, by intraperitoneal injection for 5 continuous days. The levels of serum creatinine and urea nitrogen were detected. The expression of kidney injury molecule-1 in urine was detected by enzyme-linked immunosorbent assay. The expression levels of superoxide dismutase and malondialdehyde in kidney tissue were measured. Hematoxylin-eosin staining was used to observe the pathological changes of liver tissue. The protein expression of nuclear factor E2-related factor 2 protein in kidney tissue was detected by immunohistochemistry. The protein expression of nuclear factor E2-related factor 2 and hemeoxygenase-1 protein in kidney tissue cells was detected by western blot assay.   RESULTS AND CONCLUSION: Compared with the model group, the levels of serum creatinine and urea nitrogen and urinary kidney injury molecule 1 were significantly reduced in the asiaticoside group (all P < 0.05). Asiaticoside could also significantly increase the expression of superoxide dismutase (P < 0.05), decrease the expression of malondialdehyde in kidney tissue (P < 0.05), and improve the pathological changes of kidney tissue. Compared with the asiaticoside group, the combined use of asiaticoside and bruceol could increase the levels of serum creatinine and urea nitrogen, urinary kidney injury molecule 1, and malondialdehyde in the kidney tissue, while decrease the level of superoxide dismutase in the kidney tissue (all P < 0.05). Meanwhile, asiaticoside could significantly induce the translocation of nuclear factor E2-related factor 2, and increased the expression level of hemeoxygenase-1. Overall, these findings indicate that asiaticoside can protect against kidney ischemia-reperfusion injury by activating nuclear factor E2-related factor 2/hemeoxygenase-1 pathway. Figures and Tables | References | Related Articles | Metrics

Select

Effect of moxibustion pretreatment on autophagy and NLRP3 inflammasome expression in cerebral ischemia-reperfusion model rats Jiang Jie, Zhao Baixiao, Chen Libin, Wen Li, Zhang Shanshan, Ma Jie, Zhao Hua 2022, 26 (23):  3615-3619.  doi: 10.12307/2022.658 Abstract ( 396 )   PDF (2294KB) ( 65 )   Save BACKGROUND:  Moxibustion as one of the Chinese medicine therapies has shown significant clinical efficacy in the prevention and treatment of stroke. OBJECTIVE: To explore the effect of moxibustion pretreatment on autophagy and NLRP3 inflammasome expression in rats with cerebral ischemia reperfusion.  METHODS: Thirty-six Sprague-Dawley rats were randomly divided into sham operation group, model group, and moxibustion pretreatment group, with 12 rats in each group. The moxibustion pretreatment group was given moxibustion at Baihui, Dazhui, and Zusanli before modeling. Each acupoint was given moxibustion for 3 strengths, once a day for 7 days. In the model and moxibustion pretreatment groups, the rat model of middle cerebral artery occlusion was made by suturing of the middle cerebral artery 30 minutes after the last moxibustion. After 2 hours of cerebral ischemia, the middle artery suture was removed and the rats were reperfused for 12 hours. In the sham operation group, only the common carotid artery, internal carotid artery, and external carotid artery were dissected without suturing the middle cerebral artery. The neurological deficit score was assessed, the cerebral infarct volume was calculated using 2,3,5-triphenyltetrazolium chloride staining, and the expression levels of autophagy-related proteins Beclin1, p62 and NLRP3 inflammasome in cerebral cortex ischemic area were detected using western blot assay.  RESULTS AND CONCLUSION: The neurological deficit score in the model group was significantly higher than that in the sham operation group (P < 0.01), and the neurological deficit score of the moxibustion pretreatment group was significantly lower than that of the model group (P < 0.01). 2,3,5-Triphenyltetrazolium chloride staining results showed that there were no obvious infarcts in the brain tissue of rats in the sham operation group, and obvious ischemic lesions were detected in the right brain tissue of rats in the model and moxibustion pretreatment groups. The infarct volume in the moxibustion pretreatment group was significantly less than that in the model group (P < 0.01). Western blot results showed that compared with the sham operation group, the protein expression of Beclin1 and NLRP3 was significantly increased in the model and moxibustion pretreatment groups (P < 0.01), while the expression of p62 protein decreased (P < 0.01). Compared with the model group, the expression of Beclin1 protein was significantly increased (P < 0.05), and the expression of p62 and NLRP3 proteins was significantly decreased in the moxibustion pretreatment group (P < 0.01). To conclude, moxibustion pretreatment can significantly improve the neurological function of rats after cerebral ischemia-reperfusion, and its mechanism may be related to the activation of autophagy and inhibition of the expression of inflammatory factors. Figures and Tables | References | Related Articles | Metrics

Select

Salbutamol attenuates hyperoxia-induced acute lung injury in rats Mei Hong, Qin Song, Li Kang, Chen Miao 2022, 26 (23):  3657-3663.  doi: 10.12307/2022.664 Abstract ( 355 )   PDF (27664KB) ( 28 )   Save BACKGROUND: Hyperoxia-induced acute lung injury has attracted more and more attention in clinical practice. Studies have shown that salbutamol can reduce inflammation in rats and improve the activity of various antioxidants and microcirculation, thereby reducing pulmonary edema, and exerting a protective role in lung tissue.  OBJECTIVE: To investigate the effect of salbutamol on relieving hyperoxia-induced acute lung injury and its mechanism.  METHODS: Sprague-Dawley rats were divided into a control group, a model group in which lung injury model was prepared with high concentration oxygen at different time, and a treatment group given different dosages of salbutamol by gavage (0.025, 0.05, 0.1, 0.2, 0.4 mg/kg). The optimal high concentration oxygen (> 95%) treatment time and dosage of salbutamol were determined by measuring various biochemical indicators. Real-time quantitative PCR, western blot, and hematoxylin-eosin staining were then used to determine the mRNA and protein expressions in JNK/PI3K/Akt/NF-κB/ICAM-1 signaling pathway and the pathological changes of lung tissue in the model rats with lung injury given the optimal dose of salbutamol.  RESULTS AND CONCLUSION: The hyperoxia-induced acute lung injury model was successfully constructed and showed the best quality when treated with high concentration oxygen at 60 hours. The optimal dosage of salbutamol was 0.2 mg/kg. Compared with the control group, the biochemical indicators showed a significant increase or decrease in the model group (P < 0.05). The mRNA and protein expression levels of JNK/PI3K/Akt/NF-κB/ICAM-1 signaling pathway-related factors (JNK, PI3K, Akt, nuclear factor E2 related factor 2, nuclear factor κB, intercellular adhesion molecule 1, interleukin 8, tumor necrosis factor) were significantly up-regulated in the model group compared with the control group (P < 0.05). A significant regression was observed after treatment with salbutamol (P < 0.05). Besides, the model group showed severe pathological damage and inflammatory infiltration, and salbutamol could effectively alleviate these changes in the model group. To conclude, salbutamol can significantly improve the lung injury in rats with hypoxia-induced acute lung injury through JNK/PI3K/Akt/NF-κB/ICAM-1 signaling pathways. Figures and Tables | References | Related Articles | Metrics

Select

Relationship between electroacupuncture-induced neuroprotection and neuregulin-1/epidermal growth factor receptor 4 signaling pathway in ischemia-reperfusion model rats Yao Qipeng, Liao Min 2022, 26 (23):  3664-3669.  doi: 10.12307/2022.665 Abstract ( 394 )   PDF (5456KB) ( 49 )   Save BACKGROUND: Electroacupuncture intervention at Quchi (LI11) and Zusanli (ST36) acupoints can play a neuroprotective effect and improve motor function in patients. However, there are relatively few in-depth studies on its mechanism of action. OBJECTIVE: To investigate the neuroprotective effect of electroacupuncture intervention in a rat model of cerebral ischemia-reperfusion injury based on the neuregulin-1/epidermal growth factor receptor 4 signaling pathway. METHODS: Ninety specific pathogen-free male Wistar rats were randomly divided into three groups (n=30 per group): a model group, a non-acupoint group, and an acupoint group. Another 30 rats were selected as a sham operation group with the separation of blood vessels in the left side of the neck. At 3 hours after modeling, in the non-acupuncture group, electroacupuncture was performed atthe non-acupoints under the axillary transverse striae of the right limb and 3 mm below the tip of the coccyx by using electroacupuncture, while in the acupuncture group, electroacupuncture was performed at Quchi (LI11) and Zusanli (ST36) acupoints for 7 days. Rats in the sham operation and model groups were not given any treatment, only grasped and fixed under the same conditions as the non-acupoint and acupoint groups. After modeling, the neurological deficit scores were used for model evaluation. The neurological deficit scores were evaluated in each group after treatment; and the regional cerebral blood flow and blood flow velocity on the ischemic side were measured. 2,3,5-Triphenyltetrazolium chloride staining was used to detect the volume of cerebral infarction. Electron microscope was used to observe the ultrastructure of neurons. TUNEL staining was used to detect the apoptosis of neurocytes. Western blot was used to detect the pathway protein expression of neuregulin 1/epidermal growth factor receptor 4. RESULTS AND CONCLUSION: Compared with the sham operation group, the neurological deficit score, cerebral infarction volume, neuronal apoptosis rate, neuregulin-1 protein level, and epidermal growth factor receptor 4 protein level were significantly increased, cerebral blood flow and cerebral blood flow velocity were significantly decreased in the model and non-acupoint groups (P < 0.05). The ultrastructure of neurons was abnormal in the model group and the non-acupoint group. However, there was no significant difference between the model group and the non-acupoint group (P > 0.05). Compared with the model group, the neurological deficit score, cerebral infarction volume, neuronal apoptosis rate were significantly decreased, and cerebral blood flow, cerebral blood flow velocity, neuregulin 1 protein level, and epidermal growth factor receptor 4 protein level were significantly increased in the acupoint group (P < 0.05). The ultrastructure of neurons was also significantly improved in the acupoint group. These results indicate that electroacupuncture at Quchi (LI11) and Zusanli (ST36) acupoints can significantly improve the state of neurological deficit and the ultrastructure of brain neurons , inhibit neuronal apoptosis, and play a neuroprotective effect in rats with cerebral ischemia-reperfusion injury. Its mechanism may be related to the regulation of neuregulin-1/epidermal growth factor receptor 4 signaling pathway. Figures and Tables | References | Related Articles | Metrics

Select

Metabotropic glutamate receptor improves post-stroke cognitive impairment in enriched environments Liu Jiayu, Yan Ping, Zhang Yu, Zhou Hongyu, Wang Xin 2022, 26 (23):  3676-3682.  doi: 10.12307/2022.667 Abstract ( 431 )   PDF (2064KB) ( 111 )   Save BACKGROUND: Enriched environment intervention can improve cognitive impairment to a certain extent. Glutamate is an excitatory amino acid closely related to cognition. The effect and mechanism of enriched environment on glutamate receptor in post-stroke mice are still unclear.   OBJECTIVE: To investigate the effect of metabotropic glutamate receptor on improving post-stroke cognitive impairment and its mechanism in enriched environments. METHODS: The mouse model of stroke was made by light embolus method, and the mice with cognitive impairment were selected through Y maze test in comparison with the sham operation group. The mice were randomly divided into three groups: sham operation+standard environment operation group (Sham+SE), post-stroke cognitive impairment+standard environment (PSCI+SE) group and post-stroke cognitive impairment+enriched environment (PSCI+EE) group. After 14 days in the corresponding environment, Y maze was used to detect the cognitive function of the frontal lobe. New object recognition experiment was used to detect the learning and memory ability of the hippocampus. Quantitative real-time PCR was used to detect the mRNA transcription level of metabotropic glutamate receptor in the frontal lobe and hippocampus, and western blot was used to detect the protein expression of metabotropic glutamate receptor, acetylated histone H3, CAMP response binding protein in the frontal lobe and hippocampus.   RESULTS AND CONCLUSION: Compared with the Sham+SE group, the Y maze spontaneous alternation rate was significantly decreased (P < 0.001), the new object exploration rate was significantly decreased (P < 0.001), metabotropic glutamate receptor mRNA levels in the frontal lobe and hippocampus were downregulated (P < 0.01), and the expression of related proteins in the frontal lobe and hippocampus was significantly decreased in the PSCI+SE group (P < 0.01). The spontaneous alternation rate, new object exploration rate, and metabolic glutamate receptor mRNA in the frontal lobe and hippocampus were significantly decreased, and mRNA transcription, frontal lobe and hippocampus related protein expression were significantly improved in the PSCI+EE group compared with the PSCI+SE group (P < 0.05), but there was still a significant difference between PSCI+EE group and Sham+SE group (P < 0.05). To conclude, enriched environment intervention is helpful to improve the memory of mice after stroke, and its mechanism may be through increasing the phosphorylation of CAMP response binding protein, acetylation homeostasis imbalance, increasing metabotropic glutamate receptor 2 mRNA transcription and protein expression, and ultimately improving cognitive function. Figures and Tables | References | Related Articles | Metrics

Select

Establishment and disease progression in a rat myocardial infarction model Yang Qian, Zhang Yiou, Jia Lili, Xie Jun, Feng Mali, Li Tingkai 2022, 26 (23):  3733-3737.  doi: 10.12307/2022.675 Abstract ( 672 )   PDF (5105KB) ( 72 )   Save BACKGROUND: Myocardial infarction is the most serious clinical type of coronary heart disease. The development of anti-myocardial ischemia drugs has become a research hotspot. Establishing an appropriate animal model and investigating its pathophysiological mechanism can provide an effective tool for pharmacodynamics evaluation in new drug development and promote the development of anti-myocardial infarction drugs for myocardial infarction.  OBJECTIVE: To evaluate the changes of cardiac function, tissue morphology and cardiomyocyte ultrastructure in rats after acute myocardial infarction.  METHODS: Thirty male Sprague-Dawley rats were randomly divided into two groups (n=15 per group): a sham surgery group and a model group. Rats in the model group were subjected to ligation of the left anterior descending coronary artery, while in the sham surgery group, the left anterior descending coronary artery was only threaded without ligation. On the 7th, 14th, and 28th days after modeling, changes of ST segment in lead II were observed by electrocardiogram, and the general changes of the heart were observed by thoracotomy. Myocardial infarction area was determined by 2,3,5-triphenyltetrazolium chloride staining. Hematoxylin-eosin staining was used for pathological observation of the heart, and Masson staining was adopted for the determination of myocardial fibrosis degree. Moreover, the cardiomyocyte ultrastructure was observed by transmission electron microscope.  RESULTS AND CONCLUSION: Compared with the sham surgery group, the results of electrocardiogram showed that the J point was obviously elevated on the 7th day, and the Q wave waveform was broadened and the amplitude was increased on the 14th day. The gross observation of the heart showed that rats in the model group had left ventricular hypertrophy on the 7th and 14th days and cardiac atrophy on the 28th day after operation. The results of 2,3,5-triphenyltetrazolium chloride staining showed myocardial infarction area in the model group was increased progressively. Hematoxylin-eosin and Masson staining results indicated that myocardial cells arranged disorderly, accompanied by myocyte necrosis, inflammatory cell infiltration, and fibrous proliferation that were gradually aggravated. Under the transmission electron microscope, the mitochondrial swelling was worsened in the cardiomyocytes, and the number of mitochondrial cristae was significantly reduced. All these findings reveal that the changes of cardiac function, tissue morphology, and ultrastructure of cardiomyocytes in the Sprague-Dawley rat model of myocardial infarction accord with the occurrence and development of myocardial infarction, which can provide experimental evidence for the development of therapeutic drugs for myocardial infarction. Figures and Tables | References | Related Articles | Metrics

Select

Preparing a rabbit model of synovitis in knee osteoarthritis based on the “injury-repair-reinjury” method Su Jianqing, Sun Bo, Ding Yunrong, Liu Guangming, Ji Wei, Jiang Enyu, Yang Jiayu 2022, 26 (23):  3738-3743.  doi: 10.12307/2022.676 Abstract ( 441 )   PDF (2731KB) ( 68 )   Save BACKGROUND: At present, surgical trauma and intraarticular injection-induced models are the animal models mainly used for synovitis in knee osteoarthritis. There is still a large distance between the modeling method and the mode of disease occurrence. OBJECTIVE: To establish and evaluate a rabbit model of synovitis in knee osteoarthritis based on the method of “injury-repair-re-injury,” so as to provide model support for the study of synovitis in knee osteoarthritis. METHODS: Sixteen New Zealand white rabbits were randomly divided into two groups: model group (n=8) and control group (n=8). The rabbit’s right knee was injected with 4% papain and 0.03 mol/L L-cysteine to establish the model of synovitis in knee osteoarthritis on days 1, 4, 7, and 12. The skin temperature of the right knee was recorded. Rabbits in each group were killed under anesthesia and the knee joint samples were collected on day 15. Knee joint effusion and cartilage were observed using magnetic resonance imaging. Synovial tissue changes were observed using hematoxylin-eosin staining. Expression levels of inflammatory factors and prostaglandin E2 in the knee synovial fluid were measured by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: In the model group, the rat knees became red, swollen and hot obviously at the first injection of the preparation, and the skin temperature was significantly higher than that in the control group. Magnetic resonance imaging revealed obvious synovial effusion and slight damage to the knee cartilage in the model group. Hematoxylin-esoin staining showed synovial tissue thickening and a large number of inflammatory cells infiltrated in the model group. Levels of inflammatory factors and prostaglandin E2 in the synovial fluid were significantly higher in the model group than the control group. Rabbit model of synovitis in knee osteoarthritis can be successfully established by the “injury-repair-reinjury” method, which presents similar conditions with clinical synovitis in knee osteoarthritis, by comprehensive evaluation based on the performance of knee joint surface, skin temperature of the knee, synovial effusion and histological changes of the synovial tissue.  Figures and Tables | References | Related Articles | Metrics

Select

Moist exposed burn ointment intervenes with wound healing and expression of alpha-smooth muscle actin in burn model rats Yang Shukai, Wa Qingbiao, Yuan Xiaoyan 2022, 26 (23):  3762-3767.  doi: 10.12307/2022.680 Abstract ( 466 )   PDF (4747KB) ( 108 )   Save BACKGROUND: Moist exposed burn ointment (MEBO) can inhibit inflammation, reduce oxidative stress, and accelerate wound healing. It has been widely used in the treatment of burn wounds, skin radiation damage, and diabetic foot disease, and has achieved significant results. However, the research on its mechanism is still in a blank stage. OBJECTIVE: To investigate the effects of MEBO on wound healing and expression of α-smooth muscle actin in a rat burn model through the transforming growth factor β1 (TGF-β1)/Smad family member 3 (Smad3) signaling pathway. METHODS: Eighty specific-pathogen free male Sprague-Dawley rats were randomly divided into a sham scald group, a model group, a MEBO group, and a TGF-β1 group. Except for the sham scald group, rats in the other groups were treated with a circular scald apparatus (2.5 cm in diameter) to construct rat models. Rats in the MEBO group were treated with MEBO. Rats in the TGF-β1 group were treated with TGF-β1. Rats in the sham scald and model groups were treated with physiological saline. On the 21st day, the rats were killed under anesthesia, and wound healing was observed in each group. Hematoxylin-eosin staining was used to observe the pathological manifestations of the rat wound. Immunohistochemical staining was used to detect the expression of epidermal growth factor receptor and α-smooth muscle actin. Enzyme-linked immunosorbent assay was used to detect the expression of inflammatory factors, tumor necrosis factor-α, interleukin-1β, and interleukin-6, in rat serum. Xanthine oxidase method and thiobarbituric acid coloration method were used to detect the levels of oxidative stress indicators, superoxide dismutase and malondialdehyde, in wound tissue. Western blot was used to detect the protein expression of TGF-β1, and phospho-Smad3 in rat wound tissue.  RESULTS AND CONCLUSION: Compared with the sham scald group, the wound healing rate and the levels of epidermal growth factor receptor, α-smooth muscle actin, superoxide dismutase, TGF-β1, and phospho-Smad3 were significantly reduced (P < 0.05); and the levels of tumor necrosis factor-α, interleukin-1β, interleukin-6, and malondialdehyde were increased significantly in the model group (P < 0.05). Compared with the model group, the wound healing rate and the levels of epidermal growth factor receptor, α-smooth muscle actin, superoxide dismutase, TGF-β1, and phosoho-Smad3 were significantly increased in the MEBO group and TGF-β1 group (P < 0.05). Epithelial tissue basically formed covering the wound surface, and the collagen tissue was clearly visible in the MEBO group and TGF-β1 group. Compared with the model group, the levels of tumor necrosis factor-α, interleukin-1β, interleukin-6, and malondialdehyde were significantly decreased in the MEBO group and TGF-β1 group (P < 0.05). However, these above-mentioned indicators showed no significant difference between the MEBO group and the TGF-β1 group (P > 0.05). To conclude, MEBO can reduce oxidative stress and inflammation in burn rats by activating the TGF-β1/Smad3 signaling pathway, and increase the expression level of α-smooth muscle actin, thereby promoting wound healing.  Figures and Tables | References | Related Articles | Metrics

Select

CRID3, a blocker of apoptosis-associated speck-like protein containing a card, influences local gene transcription in mice with acute spinal cord injury: a transcriptomic analysis Li Jinglu, Wang Sainan, Wang Yangyang, Fu Guiqiang, Wang Ying, Hu Jianguo, Tang Jie, Lyu Hezuo 2022, 26 (23):  3620-3632.  doi: 10.12307/2022.659 Abstract ( 457 )   PDF (3740KB) ( 57 )   Save BACKGROUND: Inflammasomes play an important role in spinal cord injury. Apoptosis-associated speck-like protein containing a CARD is a common adaptor protein of inflammasome. Our previous studies have shown that CRID3, a specific oligomerization blocker of apoptosis-associated speck-like protein containing a card, can inhibit the activation of inflammasome and the production of corresponding cytokines by inhibiting the oligomerization of apoptosis-associated speck-like protein containing a card, so as to improve the local microenvironment of the injured spinal cord and play a neuroprotective role. However, its effect on spinal cord injury at transcriptional level has not been reported. OBJECTIVE: To investigate the effect of CRID3 on local gene transcription at acute phase (8 hours) of spinal cord injury in mice by using RNA-sequencing.   METHODS: Thirty female C57BL/6 mice aged 8 weeks and weighing 18-20 g were divided into a sham operation group and a spinal cord injury group. Mice in the spinal cord injury group were randomly subdivided into a control group and a CRID3 administration group. Mice in the CRID3 administration group were intraperitoneally injected with CRID3 (50 mg/kg) after operation, while mice in the control group were injected with an equal volume of physiological saline solution. At 8 hours after spinal cord injury, three mice from each group were perfused, and the spinal cord was taken to make frozen sections that were then stained with hematoxylin-eosin to determine whether the spinal cord injury model was successfully established. Meanwhile, another three mice were selected from each group to taken spinal cord tissue. The total RNA was then extracted and purified for library preparation and RNA-sequencing. The differential gene expression of the three groups was analyzed by DESeq2 software. Another six mice were selected from the control group and the CRID3 administration group, and spinal cord tissue was taken to extract and purify total RNA. The results of RNA-sequencing were verified by real-time quantitative reverse transcription PCR. GOseq R and KOBAS software were used for gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis of the differentially expressed genes. Based on literature mining, signaling pathways related to inflammasomes were explored, and the effect of CRID3 on the expression level of related genes were further analyzed. String protein-protein interaction analysis was used to detect CRID3-related proteins.  RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that the spinal cord injury model was successfully constructed. The results of RNA-sequencing showed that compared with the sham operation group, there were 5 661 differentially expressed genes in the control group, including 3 427 up-regulated genes and 2 224 down-regulated genes. Compared with the control group, 2 924 differentially expressed genes were found in the CRID3 administration group, including 1 409 up-regulated genes and 1 515 down-regulated genes. The results of gene ontology analysis showed that the differentially expressed genes were mainly rich in chemokine receptor binding, G-protein coupled receptor binding, extracellular-glutamate-gated ion channel activity, excitatory extracellular ligand-gated ion channel activity, transmembrane transporter activity, and acid phosphatase activity. Kyoto gene and genome encyclopedia analysis combined with literature mining showed that the signaling pathways related to inflammasome mainly included tumor necrosis factor, Toll-like receptor, nuclear factor‑κB, PI3K-Akt, hypoxia-inducible factor-1, MAPK, NOD-like receptor signaling pathways as well as pyrocytosis, leukocyte transendothelial migration, and interaction between cytokines and receptors. The genes most sensitive to CRID3 included Asc, Casp4, Cyba, Cybb, F11r, Hif1a, Il18, Il1b, Itgal, Itgam, Itgb2, Jam3, Mmp3, Mmp9, and Tlr4. The results of String protein-protein interaction analysis showed that inflammasome components (Nlrp1, Nlrp3, Nlrp6, Nlrc4, Aim2, ASC, Csap1, and Csap4) interacted closely, and they formed a complete link with interleubin 1b and interleubin 18 through Csap8. In addition, some important nodal proteins such as Actn1, Cxcl5, Cd14, Cyba, Cybb, Fgf2, Hif1a, F11r, Itgal, Itga2b, Itgam, Itgb1, Jam3, Mmmp3, and Tlr4 were identified. To conclude, after spinal cord injury, a large number of genes related to inflammasomes can be activated and the signaling pathways formed by these genes may be the cause or result of the activation of inflammasomes. CRID3 can directly or indirectly inhibit gene expression in the acute stage of spinal cord injury by inhibiting the expression of inflammasome-related genes. Its related molecules and signaling pathways are mainly related to inflammatory response, local hypoxia, cell adhesion, migration, differentiation, proliferation and apoptosis. CRID3 can improve the local microenvironment of spinal cord injury in the acute stage, and these key node molecules can also be used as new therapeutic intervention targets. Figures and Tables | References | Related Articles | Metrics

Select

Salvianolic acid A effects on hippocampal protein expression in ischemic stroke rats: a tandem mass tag-based proteomic analysis Xu Wenshan, Jiang Pingli, Liu Yulu, Ding Yanyi, Yu Yan, Yang Minguang, Liu Weilin, Chen Lidian 2022, 26 (23):  3714-3720.  doi: 10.12307/2022.672 Abstract ( 469 )   PDF (3048KB) ( 54 )   Save BACKGROUND: It is unclear about the effect of salvianolic acid A on hippocampal protein expression after ischemic stroke and its mechanism.  OBJECTIVE: To observe the effect of salvianolic acid A on differential protein expression in the hippocampus of ischemic stroke rats, and to explore its possible neuroprotective mechanism.  METHODS: The modified Longa method was used to prepare a rat model of ischemic stroke. After modeling, the rats were randomly divided into normal saline group (n=6) and salvianolic acid A group (n=6). Salvianolic acid A (2 mg/kg) was injected through the tail vein after the cord plug was pulled out. The normal saline group was injected with the equal volume of normal saline. Neurological function of the rats was evaluated by the modified neurological deficit score. T2-weighted imaging was used to detect infarct volume. Open field test and Morris water maze test were used to observe the autonomous behavior, learning and memory ability of rats in each group. Tandem mass tag-based quantitative proteomics technique was used to analyze differential protein expression in the ischemic hippocampus of ischemic stroke model rats.  RESULTS AND CONCLUSION: Compared with the normal saline group, the modified neurological deficit score of rats was significantly decreased in the salvianolic acid A group. T2-weighted imaging results showed that the cerebral infarction volume in the salvianolic acid A group was significantly lower than that in the normal saline group. Results from the open field test showed that the rats in the salvianolic acid A group had better performance of autonomous activity and free exploration. Morris water maze test results showed that in the directional navigation experiment, the escape latency was significantly shortened and the number of platform crossings was significantly increased in the salvianolic acid A group compared with the normal saline group. Analysis of differentially expressed proteins in the ischemic hippocampus revealed that 50 differentially expressed proteins were up-regulated and 3 differentially expressed proteins were down-regulated between salvianolic acid A group and normal saline group. Gene ontology analysis showed that these differentially expressed proteins were mainly involved in several molecular functions such as binding and catalytic activity. Kyoto Encyclopedia of Genes and Genomes analysis showed that these differentially expressed proteins were mainly involved in several signaling pathways, such as complement and coagulation cascades, and Staphylococus aureus infection. All these findings indicate that salvianolic acid A may play a neuroprotective role in ischemic stroke rats by regulating differentially expressed proteins and complement and coagulation cascade signaling pathways in the ischemic hippocampus.    Figures and Tables | References | Related Articles | Metrics

Select

Isolation and identification of fibroblast-like synoviocytes from tree shrews and establishment of TLR8 pathway related molecular detection methods Zhou Fanqi, Li Baoying, Wang Kaitao, Tan Quanquan, Yun Chenxia, Leng Jing 2022, 26 (23):  3721-3727.  doi: 10.12307/2022.673 Abstract ( 341 )   PDF (6605KB) ( 51 )   Save BACKGROUND: Fibroblast-like synoviocytes play an important role in the pathological process of rheumatoid arthritis. When inflammatory arthritis occurs, fibroblast-like synoviocytes in the synovial lining layer will proliferate in large quantities, and the release of inflammatory factors during this period can cause joint inflammation and cartilage damage.  OBJECTIVE: To establish methods for isolation, culture, purification, and identification of tree shrew knee joint fibroblast-like synoviocytes in vitro, and explore their biological characteristics; to establish methods for detection of TLR8 pathway related molecules in fibroblast-like synoviocytes from tree shrews, and preliminarily explore the activation of TLR8 pathway in the cells.  METHODS: Fibroblast-like synoviocytes from tree shrews were isolated and purified using tissue block method and continuous passage method. Cell proliferation was detected using cell counting kit-8. Cell morphology was observed using hematoxylin-eosin staining. Immunocytochemical staining was used to detect the expression of vimentin in synoviocytes. Before and after stimulation with human TLR8 ligand R848, the expression of TLR8 and its signaling pathway proteins in tree shrew fibroblast-like synoviocytes and human fibroblast-like synoviocytes was detected using western blot assay, while the expression of TLR8 pathway and its downstream mRNAs in tree shrew fibroblast-like synoviocytes were detected using RT-PCR. RESULTS AND CONCLUSION: Under an inverted microscope, tree shrew fibroblast-like synoviocytes after 3 generations of subculture were mostly spindle-shaped cells with similar size, high purity, good growth and proliferation in vitro. Hematoxylin-eosin staining revealed that the cells were mostly spindle-shaped, similar in size. Immunocytochemical staining results showed that Vimentin positively expressed in fibroblast-like synoviocytes. Cell counting kit-8 testing results showed an “S”-shaped growth curve of tree shrew fibroblast-like synoviocytes. Western blot results revealed that at 48 hours after R848 stimulation, the expression level of nuclear factor-κB protein significantly increased (P < 0.05), the expression level of p-P38 protein decreased (P < 0.05), and there was no change in the expression of TLR8, MyD88, P38, p-Erk1/2, and RANKL proteins (in tree shrew fibroblast-like synoviocytes (P > 0.05). After R848 stimulation, the expression of p-P38 in human fibroblast-like synoviocytes also significantly decreased, and no changes were found in the expression of TLR8, MyD88, P38, p-Erk1/2, and RANKL proteins (P > 0.05). Findings from RT-PCR and agarose gel electrophoresis detection showed that the mRNA expression of tumor necrosis factor ɑ, interleukin 6, interferon β, TLR9-2, TLR9-1, TLR8-2, TLR8-1, TLR7-2, and TLR7-1 mRNA expression had no significant change after 48 hours of stimulation with R848 (P > 0.05). To conclude, the tissue block adherence method can be used to isolate tree shrew fibroblast-like synoviocytes, and TLR8 pathway-related molecular detection method for tree shrew fibroblast-like synoviocytes is successfully established.  Figures and Tables | References | Related Articles | Metrics

Select

CTLA4.FasL promotes the engraftment of xenogeneic hepatic oval cells in rat spleen Wan Zhen, Wang Xuzhen, Zhang Xiaogang 2022, 26 (23):  3728-3732.  doi: 10.12307/2022.674 Abstract ( 484 )   PDF (3217KB) ( 79 )   Save BACKGROUND: Cytotoxicn lymphocyte antigen 4.fasrymndrit ligand (CTLA4.FasL) can effectively inhibit alloreactive and autoreactive T lymphocytes. However, the inhibitory effect of CTLA4.FasL on xenogeneic T lymphocytes in hepatic oval cell transplantation via the spleen remains unclear.   OBJECTIVE: To investigate the effect of CTLA4.FasL on the proliferation of xenogeneic lymphocytes, and to monitor the survival and engraftment of CTLA4.FasL-hepatic oval cells in rat spleen. METHODS:  The recombinant lentiviral vectors carrying CTLA4.FasL fusion gene (Lv/CTLA4.FasL) and polybrene were added to the rat hepatic oval cell suspension. We used hepatic oval cells that were not infected with lentivirus and infected blank lentivirus-hepatic oval cells as controls, and then observed the viability of hepatic oval cells and the expression of red fluorescent protein lentiviral vector under a fluorescence microscope. Sprague-Dawley rats undergoing 2/3 hepatectomy were transplanted with CTLA4.FasL-hepatic oval cells, blank lentivirus-hepatic oval cells, hepatic oval cells, or phosphate buffered saline (without hepatic oval cells) through the spleen. At 1, 5, 14, and 21 days after hepatectomy, the concentration of CTLA4.FasL in rat serum was determined by enzyme linked immunosorbent assay, and the spleen of the recipient rats was taken for CK-19 immunohistochemical staining. C57BL/6 mouse lymphocytes were used as stimulator cells, and Sprague-Dawley rat lymphocytes as response cells. In the mixed lymphocyte reaction, CTLA4.FasL-hepatic oval cells, blank lentivirus-hepatic oval cells, and hepatic oval cells were inoculated, or CTLA4.FasL-hepatic oval cells (a mass concentration  of 10, 50, and 100 μg/L), blank lentivirus-hepatic oval cells, and hepatic oval cell supernatant. After 96 hours of culture, 5-bromodeoxyuridine method was used to detect the proliferation of lymphocytes in each group.   RESULTS AND CONCLUSION: Both CTLA4.FasL-hepatic oval cells and the culture supernatant could effectively inhibit the proliferation of rat lymphocytes in the xenogeneic mixed lymphocyte culture system. Spleen lymphocytes from the recipient rats transplanted with CTLA4.FasL-hepatic oval cells had a low immune response to mouse lymphocytes, while the spleen cells derived from the recipient rats transplanted with blank lentivirus-hepatic oval cells and hepatic oval cells showed strong responses. CK-19 positive cells were detected in rat spleen parenchyma of the CTLA4.FasL-hepatic oval cell group but not in the phosphate buffered saline group, hepatic oval cell group, and blank lentivirus-hepatic oval cell group at 21 days after transplantation. The results show that CTLA4.FasL can promote the survival and engraftment of hepatic oval cells in the rat spleen parenchyma through inhibiting the proliferation of xenogeneic lymphocytes. Figures and Tables | References | Related Articles | Metrics

Select

Comprehensive analysis of competing endogenous RNA network and potential biomarkers in alcohol-induced osteonecrosis of the femoral head Qian Xiaofen, Zeng Ping, Liu Jinfu, Chen Cai, Fan Siqi, Nong Jiao, Deng Wei 2022, 26 (23):  3670-3675.  doi: 10.12307/2022.666 Abstract ( 385 )   PDF (2029KB) ( 71 )   Save BACKGROUND: Alcohol-induced osteonecrosis of the femoral head is highly correlated with alcohol consumption. However, its specific pathogenesis is still unclear and there are certain individual differences. OBJECTIVE: To investigate the potential pathogenesis of alcohol-induced osteonecrosis of the femoral head by constructing a gene network map of alcohol-induced osteonecrosis of the femoral head. METHODS: Three patients with alcohol-induced osteonecrosis of the femoral head and three healthy volunteers were recruited from the Second Department of Orthopedics in the First Affiliated Hospital of Guangxi University of Chinese Medicine. Differentially expressed mRNA and lncRNA were screened out through gene sequencing of peripheral blood samples, to predict the interaction targets of lncRNA-miRNA and miRNA-mRNA. The competitive endogenous RNA regulatory network was constructed using cytoscape software. RESULTS AND CONCLUSION: Differential expression analysis identified 127 differentially expressed lncRNAs and 921 differentially expressed mRNAs from the samples of alcohol-induced osteonecrosis of the femoral head. Then, through target gene prediction, 205 miRNAs and 5 184 mRNA targets were predicted, and finally the competing endogenous RNA network containing 3 lncRNAs (SNHG3, MUC19, LINC00476), 5 miRNAs (hsa-miR-146b-5p, hsa-miR-139-5p, hsa-miR-126-3p, hsa-miR-193a-3p, hsa-miR-135a-5p), and 11 mRNAs (PEX5, ACTR3B, CHMP4B, CSMD1, MTRF1, ZNF3, HTR7, NR5A2, SYT14, CASK, ING5) was constructed, involved in stem cell differentiation, inflammatory response, neuroendocrine, gene polymorphism, and other biological processes. To conclude, a series of potential biomarkers of alcohol-induced osteonecrosis of the femoral head have been screened out by constructing the competing endogenous RNA network, which provides new ideas for further research on its molecular mechanism and therapeutic targets. Figures and Tables | References | Related Articles | Metrics

Select

Screening and biological function analysis of inflammation-related circRNAs in synovial tissue of patients with primary knee osteoarthritis Qiao Linghui, Yuan Tao, Han Jie, Wang Guancheng, Gu Yanglin 2022, 26 (23):  3683-3690.  doi: 10.12307/2022.668 Abstract ( 439 )   PDF (12449KB) ( 34 )   Save BACKGROUND: Primary knee osteoarthritis is one of the most common chronic diseases among the elderly, which brings a heavy burden on society and families. Current clinical treatments only focus on surgeries such as total knee replacement, but it is difficult to prevent cartilage tissue degeneration at an early stage. The formation mechanism of knee osteoarthritis, especially the influence of inflammation in disease progression, has been reported to some extent, but its specific mechanism is still unclear. OBJECTIVE: To investigate the differentially expressed circRNA sites of primary knee osteoarthritis and rheumatoid arthritis and their effects on the pathogenesis of the disease.  METHODS: In this study, we collected the synovial tissues from eight patients with primary knee osteoarthritis and two patients with rheumatoid arthritis (control group). The expression profile of circRNAs in the tissue was detected by RNA-seq technique, trying to find the differentially expressed genes and key biological function pathways.  RESULTS AND CONCLUSION: Compared with patients with rheumatoid arthritis, 185 differentially expressed circRNAs were detected in patients with knee osteoarthritis, of which 14 were up-regulated and 171 were down-regulated. Through the pathway enrichment and functional annotation of these target genes, a variety of enrichment pathways were identified, including protein H3-K36 dimethylation, glycosphingolipid biosynthesis process, and positive regulation of Toll-like receptor 9 signaling pathway. Based on the above sequencing results, a circRNA-miRNA interaction network was created, contributing to understanding the effect of differentially expressed circRNAs. In this study, we determined the differential expression of inflammation-related circRNAs in synovial tissue and the control group. These differentially expressed transcripts may elucidate the effect of circRNA and its relationship network in synovial tissue on the progression of osteoarthritis. Findings from this study will be helpful to explore the pathogenesis and key therapeutic targets of osteoarthritis. Figures and Tables | References | Related Articles | Metrics

Select

Bioinformatics analysis of gene expression profile of peripheral blood lymphocytes in patients with osteoarthritis Yang Wei, Yuan Puwei, Du Longlong, Li Xuefeng, Gao Qimeng, Han Qingmin 2022, 26 (23):  3706-3713.  doi: 10.12307/2022.671 Abstract ( 524 )   PDF (3973KB) ( 65 )   Save BACKGROUND: At present, there are no sensitive markers for monitoring the occurrence or progression of osteoarthritis. The detection of changes in peripheral blood gene expression profiles during the active period of osteoarthritis is helpful to investigate the accurate diagnosis and treatment targets in the blood and explain the pathogenesis.   OBJECTIVE: To analyze the differences in the gene expression profiles of peripheral blood lymphocytes between osteoarthritis patients and healthy people by bioinformatics methods, and to explore the diagnosis and treatment targets of osteoarthritis from the molecular level in the blood, so as to provide new ideas for the study of osteoarthritis. METHODS:  We searched osteoarthritis blood-related chip data from Gene Expression Omnibus and ArrayExpress databases, and downloaded the GSE63359 data set. The screening samples included 46 osteoarthritis patients’ blood and 26 healthy people’s blood, including 32 female patients and 19 healthy women. The R language limma package was used to screen the differentially expressed genes between male/female osteoarthritis patients and male/female healthy people. The ggplot2 package was used to draw volcano plots, and the ComplexHeatmap package was used to draw heat maps. The threshold was set to  P < 0.05 & |log2FC|>0.5 to obtain differentially expressed genes, and then a Venn diagram was made to obtain eight differentially expressed genes: MAP2K7, CREBZF, CLK4, TRIM37, IL18RAP, LRRN3, BLNK, and MS4A1. DAVID was used to analyze the gene ontology and Kyoto encyclopedia of genes and genomes pathways of differentially expressed genes, and the R language ggplot2 package was used to draw bubble plots. STRING and Cytoscape software were used to constructed protein-protein interaction network. Mcode and centiscape plug-in were used for module analysis, and the Cytohubba was used to screen out key genes.   RESULTS AND CONCLUSION: A total of 115 differentially expressed genes were screened out, including 16 up-regulated genes and 99 down-regulated genes. The gene ontology enrichment analysis of all differentially expressed genes mainly focused on “lymphocyte-mediated immunity,” “humoral immune response,” “antigen receptor-mediated signaling pathway,” “B cell receptor signaling pathway,” “immunoglobulin-mediated immune response,” “positive regulation of phagocytosis,” and other biological functions. Kyoto encyclopedia of genes and genomes was mainly enriched in five pathways related to osteoarthritis: hematopoietic cell lineage, Th1 and Th2 cell differentiation, Th17 cell differentiation, osteoclast differentiation and TNF signaling pathway. Protein-protein interaction network and related plug-ins were used to screen out 10 key genes, including 8 core genes, that were highly related to osteoarthritis: tumor necrosis factor, CD19, transferrin receptor, pairing box 5, mitogen-activated protein kinase 7, CD24, CD20 and B cell connection, which were highly related to osteoarthritis inflammation and cell apoptosis. The bioinformatics analysis indicates that the differences in peripheral blood lymphocytes cells gene expression between osteoarthritis patients and healthy people are concentrated in cell apoptosis and inflammation, and thus blood expression profile becomes an effective breakthrough for monitoring osteoarthritis target markers and studying its potential molecular mechanisms. Figures and Tables | References | Related Articles | Metrics