【作者】 毕荣修；

【导师】 黄桂成；

【作者基本信息】 南京中医药大学 ， 中医骨伤科学， 2010， 博士

【摘要】 膝骨性关节炎(KOA)是引起老年人膝关节疼痛和伤残的主要原因,其所致不适以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治疗该病针对性强,疗效显著,且远期效果好,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,建立动物实验模型并进行了临床研究,探讨了KOA的发病机理和中医药治疗KOA的有效性和可行性,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：观察关节止痛胶囊对实验性膝骨性关节炎的血清、关节液生化指标、自由基水平、细胞因子以及软骨细胞、软骨基质和滑膜的影响,并探讨其作用机理、发病机制和临床疗效。方法：随机将新西兰兔分为四组,A正常组、B模型组、C对照组、D治疗组。造模后对血清、关节液SOD、NO及自由基水平、细胞因子、软骨和滑膜iNOS含量等进行含量测定,对软骨细胞、软骨基质、滑膜等采用细胞形态学、软骨生化、软骨细胞核DNA含量图像分析、TUNEL法观察软骨细胞凋亡等观察指标加以分析；将符合纳入观察标准的120例膝骨性关节炎患者按随机化原则分为2组,分别为关节止痛胶囊治疗组60例、壮骨关节丸对照组60例。4周为一疗程,服药12周(三个疗程)后观察疗效并评价治疗效果。结果：通过测定兔膝血清、关节液的SOD、NO、IL-1、TNF-α、MMP-1及软骨和滑膜的iNOS含量表明,关节止痛胶囊可降低兔关节内NO、iNOS、IL-1、TNF-α和氧自由基的含量及MMP-1的活性,增加关节内SOD的含量,与对照组比较有显著性差异(P<0.05)。兔软骨细胞核DNA含量的图像分析表明关节止痛胶囊可以明显减少兔软骨细胞核内异染色质的数量,与对照组比较无显著性差异(P>0.05)；通过软骨生化指标的测定,显示关节止痛胶囊具有促进兔软骨基质合成,抑制其降解的作用,与对照组比较无明显差异(P>0.05),结合Masson染色结果证实关节止痛胶囊亦具有阻止兔软骨内胶原转型的作用。末端原位标记、琼脂糖凝胶电泳结果显示关节止痛胶囊能减轻内源性DNA氧化性损伤,减少DNA键断裂及低分子片段数量,减少兔软骨细胞凋亡数目；组织学证实治疗组可以阻止关节软骨胶原类型的改变,软骨有明显的修复。临床研究表明,治疗组显效率52.63%,总有效率84.21%,对照组显效率27.59%,总有效率68.97%,两者比较有显著性差异(P<0.05)；临床症状的改善,对于提高患者的自理能力和社会功能活动能力,提高生存质量,具有非常重要的意义。结论：研究结果表明,关节局部NO、iNOS、IL-1、TNF-α、MMP-1和氧自由基及是引起膝骨性关节炎的重要介质；软骨细胞凋亡过盛可能是关节软骨退变的重要原因之一,而内源性DNA氧化性损伤是其分子病理机制；软骨细胞形态学、软骨基质的改变是膝骨性关节炎的主要病理改变。关节止痛胶囊治疗膝OA的作用机制可能是提高机体免疫力,改善软骨细胞功能,促进软骨修复；抑制滑膜炎性改变,延缓软骨退变；改善微循环与血液流变学,降低骨内压；降低关节液NO、IL-1、TNF-α的含量及MMPs的活性,降低软骨和滑膜iNOS的表达,清除氧自由基,提高SOD的活性,从而起到延缓软骨的退变,促进软骨修复作用。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 having adverse effects. According to TCM syndrome differentiation theory, the overall concept and the guidance of Chinese medicine treatment of the disease targeted significant effect, and long-term effects, 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 establishment of animal models and clinical studies carried out to explore the pathogenesis of the KOA and Chinese medicine treatment of KOA’s effectiveness and feasibility of prevention and treatment. for Chinese medicine, KOA provide reliable experimental data and solid theoretical basis.Objective:To observe the Joint pain relief capsule in experimental knee pain of osteoarthritis of serum, synovial fluid biochemical parameters, free radicals, cytokines, and cartilage cells, cartilage matrix and synovial effects and to explore its mechanism of pathogenesis and clinical efficacy. Methods:New Zealand rabbits were randomly divided into four groups, A normal group, B model group, C control group, D treatment group. After modeling of the serum, the content of synovial fluid SOD, NO and free radicals, cytokines, iNOS content of cartilage and synovial membrane is to determine. cartilage cells, cartilage matrix, synovial, etc. and using cell morphology, biochemical cartilage, cartilage nuclear DNA content in image analysis, TUNEL to analyze the indicators such as observation of cartilage death; to observe the standards met the inclusion of 120 patients with knee osteoarthritis according to the principle of randomization into 2 groups, respectively, joint pain relief capsule in treating 60 patients, bolus control group,60 cases.4 weeks for a course of treatment, medication for 12 weeks (three courses) and to evaluate the effect observed after treatment.Results:By measuring the serum of rabbit knee joint fluid of SOD, NO, IL-1, TNF-α, MMP-1 and iNOS content in cartilage and synovium that joint pain can reduce the articular capsule in NO, iNOS, IL-1, TNF-a and oxygen free radical content and activity of MMP-1, increased SOD content of intra-articular, with the control group there was significant difference (P<0.05). The nuclear DNA content in rabbit cartilage image analysis showed that the Joint pain relief capsule can significantly reduce the pain of rabbit cartilage nucleus heterochromatin number compared with the control group, no significant difference (P> 0.05); by cartilage biochemical determination, showing joint pain capsules with the promotion of rabbit cartilage matrix synthesis, inhibiting its degradation in the control group showed no significant difference (P> 0.05), combined with Masson staining confirmed that joint pain capsules also have to stop rabbit cartilage collagen transformation. End in situ labeling, agarose gel electrophoresis showed that joint pain capsules can reduce endogenous oxidative DNA damage, reduced DNA cleavage and the number of low molecular weight fragments, reducing the number of apoptotic chondrocytes; histologically proven treatment group can prevent articular cartilage collagen type of change, significant cartilage repair. Clinical studies have shown that treatment group was 52.63%, 84.21% total efficiency, control group was 27.59%, total effective rate 68.97%, in comparison with a significant difference (P<0.05); clinical symptoms improved, the increase patient self-care ability and social function of activity and improve quality of life, has very important significance.Conclusion:The results showed that the joints NO, iNOS, IL-1, TNF-a, MMP-1 and oxygen free radicals are caused by knee osteoarthritis important medium; chondrocyte apoptosis excessive degeneration of articular cartilage may be the important one of the reasons The endogenous oxidative DNA damage is its molecular pathological mechanisms; cartilage cell morphology, cartilage matrix changes in knee osteoarthritis is the main pathological changes. Joint pain relief capsule in treating knee OA pain mechanism of action may improve immunity, improve cartilage cell function, promote cartilage repair; suppress synovitis of change, slow down cartilage degeneration; microcirculation and hemorheology, reduce intraosseous pressure; reduce synovial fluid NO, IL-1 and TNF-a content and activity of MMPs, reduced iNOS expression in cartilage and synovium, removing oxygen free radicals, improve the activity of SOD, which played slow the degeneration of cartilage, promoting cartilage repair. This study of Chinese medicine for the prevention and treatment of knee osteoarthritis to provide a reliable experimental data and solid theoretical basis.更多还原