【作者】 滕加文；

【导师】 许建安；

【作者基本信息】 南京中医药大学 ， 中医骨伤学， 2011， 博士

【摘要】 膝骨性关节炎(Knee Osteoarthritis KOA)是引起老年人膝关节疼痛和伤残的主要原因,它所引起的不适乃至功能障碍对病人的生活质量造成严重影响。根据中医整体观和辨证施治理论为指导的中医药疗法,对该病治疗具有针对性强、疗效显著、远期效果好等特点,可以从根本上控制病情的发展,为治本之道。本研究是在系统整理中医学对KOA认识的相关文献基础上,建立动物实验模型进行实验研究,探讨了KOA的发病机理及中药治疗机制,为中医药防治KOA提供可靠的实验资料及坚实的理论依据。目的：观察补肾壮骨胶囊治疗膝骨性关节炎的疗效,并探讨其作用机理及膝骨性关节炎的发病机制。方法：将40只新西兰兔随机分为A正常组、B模型组、C治疗组、D对照组。B、C、D三组按照Hulth造模方法造成膝骨性关节炎模型。C组给予补肾壮骨胶囊,D组给予壮骨关节丸。给药8周后拍摄膝关节正位CR片,电镜下观察软骨改变；采用苏木素-伊红(HE)染色,光镜下观察软骨病理改变,进行Mankin’s评分；采用免疫组织化学方法观察关节软骨中基质金属蛋白酶-1(MMP-1)、组织金属蛋白酶抑制剂(TIMP-1)、聚蛋白多糖酶-4(ADAMTS-4)阳性细胞率,脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测软骨细胞的凋亡指数(AI)；逆转录-聚合酶链式反应(RT-PCR)检测关节软骨细胞bcl-2 mRNA、bax mRNA表达水平。将实验数据采用SPSS16.0统计软件处理,大体评分比较采用秩和检验；组间比较用One-Way ANOVA方差分析,均数比较采用LSD、SNK-q检验,P<0.05,差异具有统计学意义。结果：在光镜和电镜下,补肾壮骨胶囊对Hulth造模方法建立的新西兰实验兔膝骨性关节炎模型关节软骨的X线影像、大体外观、组织学Mankin’s评分、软骨结构均有明显改善,对膝骨性关节炎软骨退变有明显的保护作用；补肾壮骨胶囊能降低关节软骨中MMP-1、ADAMTS-4阳性细胞率,提高关节软骨中TIMP-1阳性细胞率；补肾壮骨胶囊能降低软骨细胞的凋亡指数(AI)；提高软骨细胞bcl-2 mRNA表达,降低bax mRNA表达。结论：补肾壮骨胶囊可以抑制膝骨关节炎软骨细胞的凋亡、改善软骨病变,其作用途径可能和降低基质金属蛋白酶1(MMP-1)、聚蛋白多糖酶-4(ADAMTS-4)活性,提高基质金属蛋白酶抑制剂1 (TIMP-1)活性,上调bcl-2基因、下调bax基因有关、调节Bcl-2 mRNA/Bax mRNA的比例。本项研究为中医药防治膝骨性关节炎提供了可靠的实验资料及坚实的理论依据。更多还原

【Abstract】 Knee osteoarthritis(KOA) is the main cause of older knee pain and disability, its dysfunction due to discomfort as well as the quality of life of patients have adverse effects. According to TCM syndrome differentiation theory,the overall concept and the guidance of Chinese medicine treatment of the disease targeted significant effete,And long-term effeets,They could fundamentally control the disease’s development a permanent cure.In this study, finishing in the system knowledge of medicine to the KOA on the basis of relevant literature, the establishmnent of animal models studies carried out to explore the Pathogenesis of the KOA and Chinese medicine treatment of KOA effectiveness and feasibility of Prevention and treatment.for Chinese medicine,KOA Provide reliable experimental data and solid theoretieal basis.Objective:To observe the efficacy of Bushenzhuanggu Capsule(BSZGC) for treatment of knee osteoarthritis(KOA),and explore the mechanism in the effect of BSZGC and the mechanism of KOA.Methods:In the experiment study,40 New Zealand rabbits were randomly divided into 4 groups:Group A(the normal group), Group B (the model group), Group C(the treated group) and Group D(the control group).KOA models were induceded in Group B, C and D by Hulth’s technique. The rabbits in Group C were treated with BSZGC and Zhuangguguanjie pill (ZGGJP) was used in Group D.8 weeks later, to make CR of the knee. Observe the pathological changes of articular cartilage and by HE colouration under light microscope and scanning electron microscope, then mark it by Mankin. Immunohistochemistry was applied to observe the expression of MMP-1 and TIMP-1 and ADAMTS-4 in articular cartilages, Check chondrocyte apoptosis index(AI) by TUNEL.Measure expression rate of bcl-2 mRNA and bax mRNA by RT-PCR.The experimental data are processed with SPSS 16.0 statistical software, gross score compares the rank and examination. Comparison between groups with one-way ANOVA analysis of variance, compares the mean differences with SNK-q or LSD inspection.If P< 0.05, the differences are statistically significant. Rusult:Under light microscope and scanning electron microscope, Bushenzhuanggu Capsule could decrease macroscopic grading score, X-rayphotograph score, Mankin’s score and relief structure of articular cartilage in OA knee by Hulth’s technique,so it could prevent articular cartilage from OA degeneration. After treating with Bushenzhuanggu Capsule, chondrocyte apoptosis index(AI) were decrease,expression rate of bcl-2 mRNA were raised and that of bax mRNA were lowered. Conclusion:The Bushenzhuanggu Capsule can restrain chondrocyte apoptosis of knee Oteoarthritis,and relive damage of articular cartilage. The action may be caused by two approaches,which the first is decreasing grade of MMP-1、ADAMTS-4 and raising grade of TIMP-1, and the second is raising expression rate of bcl-2 mRNA and lowering that of bax mRNA. And adjust the ratio of Bcl-2 mRNA/Bax mRNA. This study of Chinese medicine for the prevention and treatment of knee osteoarthritis to provide a reliable experimenial data and solid theoretical basis.更多还原