中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (32): 6939-6946.doi: 10.12307/2025.939

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

蒙药沙蓬粗寡糖对小鼠滑膜细胞炎症和凋亡的影响

赵雪梅1，2，王  睿1，2，奥·乌力吉3，包书茵4，江小华1，2   

  1. 1华北理工大学基础医学院，河北省唐山市  063210；2河北省慢性疾病基础医学重点实验室，河北省唐山市  063210；3内蒙古蒙医药工程技术研究院，内蒙古自治区通辽市  028000；4内蒙古民族大学公共卫生学院，内蒙古自治区通辽市  028000


  • 收稿日期:2024-10-08 接受日期:2024-12-06 出版日期:2025-11-18 发布日期:2025-04-28
  • 通讯作者: 江小华，博士，教授，华北理工大学基础医学院，河北省唐山市 063210；河北省慢性疾病基础医学重点实验室，河北省唐山市 063210 共同通讯作者：包书茵，博士，副教授，内蒙古民族大学公共卫生学院，内蒙古自治区通辽市 028000
  • 作者简介:赵雪梅，女，1999年生，华北理工大学在读硕士，主要从事骨关节炎研究。
  • 基金资助:
    蒙药研发国家地方联合工程研究中心开放基金项目(MDK2021023)，项目负责人：江小华

Effects of Agiophyllum Oligo Saccharides on inflammation and apoptosis of mouse synovial cells

Zhao Xuemei1, 2, Wang Rui1, 2, Ao · Wuliji3, Bao Shuyin4, Jiang Xiaohua1, 2   

  1. 1School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063210, Hebei Province, China; 2Hebei Key Laboratory for Chronic Diseases, Tangshan 063210, Hebei Province, China; 3The Research Institute of Traditional Mongolian Medicine Engineering Technology, Tongliao 028000, Inner Mongolia Autonomous Region, China; 4School of Public Health, Inner Mongolia Minzu University, Tongliao 028000, Inner Mongolia Autonomous Region, China
  • Received:2024-10-08 Accepted:2024-12-06 Online:2025-11-18 Published:2025-04-28
  • Contact: Jiang Xiaohua, PhD, Professor, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063210, Hebei Province, China; Hebei Key Laboratory for Chronic Diseases, Tangshan 063210, Hebei Province, China Co-corresponding author: Bao Shuyin, PhD, Associate professor, School of Public Health, Inner Mongolia Minzu University, Tongliao 028000, Inner Mongolia Autonomous Region, China
  • About author:Zhao Xuemei, Master candidate, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063210, Hebei Province, China; Hebei Key Laboratory for Chronic Diseases, Tangshan 063210, Hebei Province, China
  • Supported by:
    Open Fund Project of National Local Joint Engineering Research Center for Research and Development of Mongolian Medicines, No. MDK2021023 (to JXH)

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摘要：


文题释义：
蒙药沙蓬粗寡糖：蒙药沙蓬是蒙医习用药材，全草可入药，主要含有黄酮类、皂苷类、生物碱、香豆素、糖类等活性成分，具有增强免疫力、降血糖、抗氧化、抗炎、抗肿瘤、保护肝脏、降低血清胆固醇、抗感染等作用。沙蓬粗寡糖是由沙蓬的干燥地上部分提取纯化而来。
Toll样受体4/核因子κB信号通路：核因子κB 信号通路可通过Toll样受体4早期激活免疫因子，从而触发滑膜释放肿瘤坏死因子α、白细胞介素1β等炎性因子，导致滑膜增生并促使骨关节炎发生。

背景：在膝骨关节炎发生发展中，炎症浸润以及成纤维细胞样滑膜细胞凋亡不足所致的异常增殖和活化是疾病进展的重要推动因素。Toll样受体4/核因子κB信号通路在调控滑膜细胞炎症反应和凋亡中起到关键作用。沙蓬粗寡糖作为一种传统蒙药成分，具有抗炎和免疫调节潜力，但具体的分子作用机制尚未被充分揭示。
目的：探讨蒙药沙蓬粗寡糖对脂多糖诱导的小鼠滑膜细胞炎症和凋亡的影响及机制。
方法：将小鼠滑膜细胞分为4组：对照组、模型组、沙蓬粗寡糖组、Bay 11-7082组，采用1.0 µg/mL脂多糖干预36 h制备骨关节炎滑膜细胞模型，沙蓬粗寡糖组加入1.0 µg/mL脂多糖和32 µg/mL沙蓬粗寡糖处理36 h，Bay 11-7082组先用1 μmol/L核因子κB通路抑制剂Bay 11-7082孵育4 h后，再加入1.0 µg/mL脂多糖和32 µg/mL蒙药沙蓬粗寡糖作用36 h。Western blot检测Toll样受体4、MyD88、NF-κBp65、p-NF-κBp65、IκBα蛋白表达，ELISA检测细胞培养基中肿瘤坏死因子α、白细胞介素1β水平，免疫荧光和Western blot检测Bax、Bcl-2蛋白表达，荧光探针DCFH-DA法检测细胞内活性氧水平。
结果与结论：①模型组肿瘤坏死因子α、白细胞介素1β水平，Toll样受体4、MyD88、Bcl-2蛋白表达，活性氧水平以及p-NF-κBp65/NF-κBp65较对照组升高(P < 0.05)，而IκBα、Bax蛋白表达较对照组降低(P < 0.05)；②沙蓬粗寡糖组肿瘤坏死因子α、白细胞介素1β水平，Toll样受体4、MyD88、Bcl-2蛋白表达，活性氧水平以及p-NF-κBp65/NF-κBp65较模型组降低(P < 0.05)，而IκBα、Bax蛋白表达较模型组升高(P < 0.05)；③Bay 11-7082组肿瘤坏死因子α、白细胞介素1β水平，活性氧水平和Bcl-2蛋白表达较沙蓬粗寡糖组升高(P < 0.05)，但仍低于模型组(P < 0.05)，Bax蛋白表达较沙蓬粗寡糖组降低(P < 0.05)，但仍高于模型组(P < 0.05)；④Bay 11-7082组Toll样受体4、MyD88蛋白表达和p-NF-κBp65/ NF-κBp65比值较模型组和沙蓬粗寡糖组降低(P < 0.05)，IκBα蛋白表达较模型组和沙蓬粗寡糖组升高(P < 0.05)。结果表明，蒙药沙蓬粗寡糖可以改善小鼠滑膜细胞炎症，促进炎症滑膜细胞凋亡，作用机制可能与调节Toll样受体4/核因子κB信号通路有关。

关键词: 骨关节炎, 沙蓬粗寡糖, 小鼠滑膜细胞, 炎症, 凋亡, 信号通路, 脂多糖, Bay 11-7082, 工程化组织构建

Abstract: BACKGROUND: For knee osteoarthritis, synovium inflammation and abnormal proliferation and activation of fibroblast-like synoviocytes caused by insufficient apoptosis are important driving factors for disease progression. Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway plays a key role in regulating inflammatory response and apoptosis of synovial cells. As a traditional Mongolian medicine component, Agiophyllum Oligo Saccharides has anti-inflammatory and immunomodulatory functions, but its specific molecular mechanism has not been fully revealed.
OBJECTIVE: To explore the effects and mechanism of Agiophyllum Oligo Saccharides on lipopolysaccharide-induced inflammation and apoptosis of mouse synovial cells.
METHODS: The mouse synovial cells were randomly divided into a control group, a model group, an Agiophyllum Oligo Saccharides group and a Bay 11-7082 group. Osteoarthritic synoviocyte model was prepared using 1.0 µg/mL lipopolysaccharide intervention for 36 hour. The Agiophyllum Oligo Saccharides group was treated with 1.0 µg/mL lipopolysaccharide and 32 µg/mL Agiophyllum Oligo Saccharides for 36 hours; and the Bay 11-7082 group was firstly incubated for 4 hours with 1 μmol/L NF-κB pathway inhibitor Bay 11-7082, and then 1.0 µg/mL lipopolysaccharide and 32 µg/mL Agiophyllum Oligo Saccharides for 36 hours. Western blot was used to detect TLR4, MyD88, NF-κBp65, p-NF-κBp65, and IκBα protein expression. The level of tumor necrosis factor-α and interleukin-1β in the supernatant were analyzed by ELISA. To detect the expressions of Bax and Bcl-2, western blot and immunofluorescence analysis were performed. The intracellular reactive oxygen species level was detected by DCFH-DA.
RESULTS AND CONCLUSION: (1) Compared with the control group, the levels of tumor necrosis factor-α, interleukin-1β and reactive oxygen species, the protein expressions of TLR4, MyD88, Bcl-2, and p-NF-κBp65/ NF-κBp65 increased (P < 0.05), whereas the protein expressions of IκBα and Bax decreased (P < 0.05) in the model group. (2) Compared with the model group, the levels of tumor necrosis factor-α, interleukin-1β and reactive oxygen species, the protein expressions of TLR4, MyD88, Bcl-2, and p-NF-κBp65/NF-κBp65 decreased (P < 0.05), while the expressions of IκBα and BAX increased (P < 0.05) in the Agiophyllum Oligo Saccharides group. (3) The levels of tumor necrosis factor-α, interleukin-1β and reactive oxygen species and the protein expression of Bcl-2 in the Bay 11-7082 group were higher than those in the Agiophyllum Oligo Saccharides group (P < 0.05), but still lower than those in the model group (P < 0.05), whereas the expression of Bax protein was lower than that in the Agiophyllum Oligo Saccharides group (P < 0.05), but still higher than that in the model group 
(P < 0.05). (4) Compared with the model group and the Agiophyllum Oligo Saccharides group, the protein expressions of TLR4, MyD88 and p-NF-κBp65/NF-κBp65 decreased (P < 0.05), whereas the protein expressions of IκBα increased (P < 0.05) in the Bay 11-7082 group. To conclude, Agiophyllum Oligo Saccharides could inhibit inflammation of mouse synovial cells and promote apoptosis of synovial cells with inflammation, and the mechanisms may be correlated with regulating the TLR4/NF-κB signaling pathway. 

Key words: osteoarthritis, Agiophyllum Oligo Saccharides, mouse synovial cells, inflammation, apoptosis, signaling pathway, lipopolysaccharide, Bay 11-7082, engineered tissue construction

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