Bushen Huoxue Decoction affects mRNA expression of various cytokines in cartilage of the rabbit models of osteoporotic osteoarthritis

doi:10.3969/j.issn.2095-4344.0371

Abstract

Abstract:

BACKGROUND: Bushen Huoxue Decoction has been found to be effective for osteoarthritis and osteoporosis, but the underlying mechanisms are little reported.
OBJECTIVE: To explore the effects of Bushen Huoxue Decoction on the mRNA expression levels of interleukin 1, interleukin 6, and transforming growth factor-β in the knee chondrocytes of female New Zealand female rabbit models of osteoporotic osteoarthritis.
METHODS: Thirty female New Zealand rabbits were randomly divided into osteoporosis, osteoarthritis, and osteoporotic osteoarthritis groups. The right knee articular cartilages were removed, and the chondrocytes were isolated and cultured after modeling. Another seven rabbits were selected to prepare seven groups of serum containing drugs: normal group, low-, medium- and high-dose Bushen Huoxue Decoction groups, glucosamine hydrochloride group, alendronate sodium group and glucosamine hydrochloride plus alendronate sodium group. The chondrocyte proliferation in each group was detected by MTT assay. The mRNA expression levels of interleukin 1, interleukin 6, insulin-like growth factor-1 (IGF-1) and transforming growth factor-β (TGF-β) in the knee chondrocytes were detected by PCR.
RESULTS AND CONCLUSION: MTT results showed that with the administration time increasing to day 7, the absorbance values in each group were significantly higher than those in the normal group (P < 0.05), except osteoarthritis and low-dose groups. Bushen Huoxue Decoction exerted effect in a concentration-dependent manner. PCR results: the order of mRNA expression level of interleukin 1 was as follows: osteoporotic osteoarthritis group > osteoarthritis group > osteoporosis group (P < 0.05). The mRNA expression level of interleukin 6 was the highest in the osteoporotic osteoarthritis group, followed by osteoarthritis group, the lowest in the osteoporosis group, and the level in the osteoporotic osteoarthritis and osteoarthritis groups was significantly higher than that in the osteoporosis group (P < 0.05). The order of mRNA expression levels of IGF-1 and TGF-β were the highest in the osteoporosis group, followed by osteoarthritis group, the lowest in the osteoporotic osteoarthritis group, and the difference was significant between osteoporosis and osteoporotic osteoarthritis groups (P < 0.05). Compared with the normal group, Bushen Huoxue Decoction significantly decreased the mRNA expression levels of interleukin 1 and interleukin 6, and significantly increased the mRNA expression levels of IGF-1 and TGF-β in a concentration-dependent manner. In summary, Bushen Huoxue Decoction can promote the proliferation of chondrocytes. The occurrence of osteoarthritis and osteoporosis is related to interleukin 1, interleukin 6, IGF-1 and TGF-β mRNA expression. There is a positive correlation between osteoarthritis and osteoporosis. Bushen Huoxue Decoction indirectly or directly protects and repairs injured chondrocytes.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoporosis, Osteoarthritis, Drugs, Chinese Herbal, Tissue Engineering

CLC Number:

R318

Chen Hai-xia1, Zhang Kai-wei2, Ma Wen-juan3, Shen Feng-jun2, Zhang Bo1. Bushen Huoxue Decoction affects mRNA expression of various cytokines in cartilage of the rabbit models of osteoporotic osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(32): 5123-5132.

Figures/Tables 6

节炎组中与正常组对比差异无显著性意义外(t=2.866，P=0.058> 0.05)，各剂量组与正常组对比差异均有显著性意义(P < 0.05)；除低、中剂量组在骨质疏松症组中对比差异无显著性意义外(t=0.916，P=0.395> 0.05)，各剂量组两两对比差异均有显著性意义(P < 0.05)，剂量与效益呈正比关系(表9)。白细胞介素1表达量OA+OP组>骨性关节炎组>骨质疏松症组，3组中白细胞介素1表达量两两比较差异均有显著性意义(P < 0.05)(图1，表10)。 2.3.2 补肾活血汤对软骨细胞内白细胞介素6影响各药物组与正常组比较，均有降低软骨细胞内白细胞介素6表达的作用，以盐+阿组与高剂量组最为明显(表11)。各剂量组与正常组对比均差异有显著性意义(P < 0.05)；除低、中剂量组在骨质疏松症组中对比差异无显著性意义(t=1.551，P=0.172> 0.05)，其他各剂量组两两对比差异均有显著性意义(P < 0.05)，且剂量与效益呈正比关系(表12)。3组中白细胞介素6表达量OA+OP组>骨性关节炎组>骨质疏松症组，骨质疏松症组与OA+OP组及骨性关节炎组对比，差异有显著性意义(P < 0.05)(图2，表13)。 2.3.3 补肾活血汤对软骨细胞内胰岛素样生长因子1的影响各药物组与正常组比较，均有上调软骨细胞内胰岛素样生长因子1表达的作用，以盐+阿组、高剂量组及盐酸组较为明显(表14)。除低剂量组在骨性关节炎组及OA+OP组中与正常组对比差异无显著性意义(P > 0.05)外，各剂量组与正常组在3组中对比差异均有显著性意义(P < 0.05)，且各剂量组两两对比差异均有显著性意义(P < 0.05)，剂量与效益呈正比关系(表15)。3组中胰岛素样生长因子1表达量骨质疏松症组>骨性关节炎组>OA+OP组，骨质疏松症组与OA+OP组对比差异有显著性意义(P < 0.05)(图6，表16)。"

References

[1] 廖德发.我国骨性关节炎流行病学调查现状[J].微创医学, 2017,12(4): 521-524.

[2] Huang W, Liu YS.Main points of diagnosis and treatment of senile osteoporosis.Chin J Geriatr.2005;24 (12):939-941.

[3] Hulet A,JP Sabatier,D Souquet, et al. Distribution of bone mineral density at the proximal tibia in knee osteoarthritis. Calcif Tissue Int 2002;71 (4): 315-322.

[4] 马玉环,郑文伟,林平冬,等.骨关节炎软骨退变与炎症的关系[J].风湿病与关节炎,2015,4(8):50-53.

[5] 夏婷,李双庆.炎症相关骨质疏松症的发病机制[J].中国骨质疏志, 2015, 21(1):117-120.

[6] 白斌,李燕琴,李曙明,等.丹参对兔骨性关节炎关节软骨白介素-1β、白介素-6和胰岛素样生长因子-I表达的影响[J].中国康复医学杂志, 2015,30(11): 990-994.

[7] 刘传文,赵蕾,滕学仁,等.金天格胶囊对膝骨关节炎患者关节液中MMP-3、TIMP-1、IL-1β、TGF- β1水平的影响[J].中国骨质疏松杂志,2015, 21(4):442- 446.

[8] Ahn JK,Huang B,Bae EK,et al.The role of α-defensin-1 and related signal transduction mechanisms in the production of IL-6, IL-8 and MMPs in rheumatoid fibroblast-like synoviocytes. Rheumatology (Oxford). 2013;52(8):1368-1376.

[9] Shen J, Li S, Chen D, TGF-beta signaling and the devel-opment of osteoarthritis．Bone Res.2014;2. pii: 14002

[10] 罗瑞,张瑞智,龚正林,等.IGF-1对体外培养髁突软骨细胞增殖与凋亡的影响[J].实用口腔医学杂志，2016，32(6):810-814

[11] 刘日光,赵瑜,沈冯君,等. 两种不同补肾方法治疗去卵巢大鼠骨质疏松症的对比研究[J].中国中医骨伤科杂志, 2003, 11(1):12-15.

[12] 黄继汉,黄晓晖,陈志扬,等. 药理试验中动物间和动物与人体间的等效剂量换算[J].中国临床药理学与治疗学,2004,9(9):1069-1072.

[13] 阿曼古力•拜哈提,杨阳. 膝关节退行性骨关节病与骨质疏松的相关关系研究 [J].世界最新医学信息文摘 ,2017,17(42):06-07.

[14] Ciner M,Monluya MJ,Vaulucv MA,et al.Diffcrences in osteogenie and apoplotir genes between osteoporotic and osteoarthntic patienls. BMC Musculoskelet Disord. 2013;14:41.

[15] 刘康妍.绝经后妇女骨质疏松症与骨关节炎的相关性研究[J].中华关节外科杂志,2012,12(6):49-51.

[16] Papaloucas CD, WardR J, Tonk in CJ, et al. Cancellous bone changes in hip osteoarthritis: a short term longitudinal study using fractal signature analysis. Osteoarthritis Cartilage, 2005, 13: 998-1003.

[17] Hayami T, Pickarski M, Zhuo Y, et al. Characterization of articular cartilage and subchondral bone changes in the rat anterior cruciate ligament transection and meniscectomized models of osteoarthritis. Bone.2006;38(2):234-243.

[18] 许学猛,王羽丰,邓晋丰,等.补肾活血胶囊影响兔膝关节退变性疾病骨内压变化的实验研究[J].中国中医骨伤科杂志. 2001,9(4):24-27.

[19] 臧洪敏,陈君长,刘亦恒,等.补肾活血方对成骨细胞增殖等生物学特性影响的实验研究[J].中药材, 2005, 28(9) :803-805

[20] 梁平,李胜利,方苏亭,等.骨炎痛定方对实验性兔膝骨关节病血清中NO和SOD含量的影响[J].中国中医骨伤科杂志, 2006,14(12):50-53.

[21] 程孟春,刘艳秋,王莉,等. 何首乌和菟丝子对破骨细胞和成骨细胞增殖及分化的影响[J]. 中国中药杂志, 2011,36(19):2737-2740.

[22] Li Y,Wang MJ,Li S,et al. Effect of total glycosides from Eucommia ulmoides seed on bone microarchitec-ture in rats.Phytother Res. 2011; 25(12):1895-1897.

[23] Lopez-Armada MJ, Carames B, Lires-Dean M. Cytokines tumor necrosis factor alpha and inter leuk in-1β,differentially regulate apoptosis in osteoarthritis cultured human chondrocytes. Osteoarthritis Cartilage. 2006;14 (7):660-669.

[24] Buvanendran A,Kroin JS,Berger RA,et al.Up regulation of prostagland in E2 and inter leukins in the central nervous system and peripheral tissue during and after surgery in humans.Anesthes iology.2006; 104(3):403.

[25] Bjornsson GL,Thorsteinsson L,Gudmundsson KO, et al. Inflammatory cytokines in relation to adrenal response following total hip replacement.Scand J Immunol.2007; 65 (1): 99-105

[26] 何启荣,林伟.关节液中TNF-α、IL-1、M M PS水平对膝骨性关节炎患者的预后的评估价值[J].中国医学创新, 2016, 7:1-4

[27] Ding M,Dalstra M,Linde F, et al. Changes in the stiffness of the human tibial cartilage bone complex in early stage osteoarthrosis. Acta Orthop Scand.1998; 69(4):358 -362.

[28] Barta E,Maroudas A.A theoretical study o f the distribution of insulin-like growth factor in human articular cartilage.Theor Biol.2006; 241(3):628-631

[29] Sakimura K, Matsumoto T, Miyamoto C, et al .Effects of insulin-like growth factor I on transforming growth factor beta l induced chondrogenesis of synovium derived mesenchymal stem cells cultrued in a po-lyglycolic acid scaffold.Celles Tissues Organs.2006;183(2): 55-61.

[30] Blaney Davidson EN, vander Kraan PM, vanden Berg WB.TGF-beta and osteoarthritis. Osteoarthritis Cartilage.2007;15(6) : 597-604.

[31] Patil AS, Sable RB, Kothari RM. An update on transforming growth factor-β (TGF-β): sources , types,functions and clinical applicability for cartilage /bone healing.J Cell Physiol.2011;226(12):3094-3103.

[32] Van Der Kraan PM．Age-related alterations in TGF beta signaling as a causal factor of artilage degeneration in osteoarthritis. Biomed Mater Eng.2014;24: 75-80.