乙酰水杨酸联合双氯芬酸对兔骨关节炎模型关节软骨影响的实验研究

发布时间：2018-03-19 10:25

本文选题：乙酰水杨酸联合双氯芬酸　切入点：骨关节炎　出处：《山东大学》2016年博士论文　论文类型：学位论文

【摘要】：骨关节炎(osteoarthritis, OA)是一种慢性疾病,关节软骨的退变和骨质增生是其常见的表现,一些活动多、磨损大、承受压力较大的关节部位较早受累。OA发病率逐年增加,患者的医疗费用年年递增,尤其是老年人关节肌肉组织退化导致外伤比例也越来越高,膝关节OA成为老年人致残的头号杀手,从而使OA逐渐由医学问题进展到社会问题,在老龄化社会的今天,OA成为影响公共健康的重点疾病。中华医学会骨科学分会制定的“骨关节炎诊治指南”中,治疗骨性关节炎的目标就是减轻关节疼痛,抑制关节的进一步损坏,尽可能地保留及提高关节的运动功能,使得患者生活质量能够大幅度提高,以非药物与药物联合的治疗方法为主常用药物包括非甾体类消炎药(NSAIDS)、止痛药、黏弹性补充药物、氨基葡萄糖等。各种药物治疗只能缓解症状,因此目前研究的重点是通过靶向OA的各种生物学和生物力学因素来最大程度减缓OA病程。大量研究表明,OA病理机制是各种原因导致的关节软骨细胞凋亡和胞外基质降解而引起关节系统损伤疾病。治疗靶点减缓或者逆转软骨细胞凋亡。而软骨细胞和滑膜细胞高表达基质金属蛋白酶系(MMPs),该酶类与基质金属蛋白酶组织抑制(TIMPs)之间的动态平衡被打破,在力学负荷作用和炎性因子的影响等共同作用下,出现软骨中基质成分的游离、蛋白多糖、软骨胶原的持续性破坏,最终导致软骨的降解和破坏。FDA在1999年发布了骨关节炎研究指南,其中包括了动物模型的应用。建立OA模型的目的是重现关节疾病,研发潜在的治疗方法。在本研究中,选择新西兰大白兔,复制人类外伤后OA模式,建立兔骨关节炎模型。对建模成功的动物进行药物治疗。实验中选用的药物有两种,乙酰水杨酸和双氯芬酸钠。乙酰水杨酸是一种解热镇痛药,可用于骨关节炎和风湿性疾病,还能抑制血小板聚集。双氯芬酸,属于非甾体抗炎药。自20世纪60年代开始,就有两种药物联合治疗骨关节炎以及类风湿性关节炎的报道,但是,对联合用药的机理以及治疗靶点,都没有详细的报道。本研究推测,OA发病过程中,涉及MMP、TIMPs、IL-1β、 NO等因子的变化,拟对OA治疗过程中几个细胞因子的活性及表达进行相关研究。本研究将分为五部分。1. Hulth法建立兔骨性关节炎模型的实验研究2.乙酰水杨酸联合双氯芬酸对兔关节炎软骨组织学方面的影响3.乙酰水杨酸联合双氯芬酸对兔关节炎关节滑液中IL-1β、NO含量的影响4.乙酰水杨酸联合双氯芬酸对兔关节炎软骨细胞中MMP-3、MMP-13表达的影响5.乙酰水杨酸联合双氯芬酸对兔关节炎软骨细胞中TIMP-1表达的影响研究目的：探讨不同浓度乙酰水杨酸联合双氯芬酸对兔骨关节炎模型关节软骨影响的实验研究。方法：乙酰水杨酸联合双氯芬酸按照不同浓度分成三组5mg/kg,10mg/kg, 20mg/kg,将32新西兰大白兔建立兔骨关节炎模型,对各组大白兔进行评分,利用试剂盒测定各组关节液中的NO. IL-1β含量,WB法测定MMP-3, MMP-13的蛋白的表达,PCR法测定TIMP-1的mRNA表达,观察各项指标的变化。结果：不同浓度的药物对兔骨关节炎有很好的治疗作用,能够降低评分分数,降低NO、IL-1β的含量,WB法可以看到MMP-3, MMP-13的蛋白表达下调,PCR法发现TIMP-1的mRNA表达上调。结论：不同浓度的乙酰水杨酸联合双氯芬酸能够有效抑制兔骨关节炎的发展,并能抑制NO、IL-1β、MMP-3、MMP-13,同时增加TIMP-1的表达,进而起到治疗作用。通过对OA治疗过程中几个细胞因子的活性及表达进行相关检测,本研究对两者联合用药的分子机制进行了探讨,从而为联合治疗奠定了坚实基础。而且这两种药物都是质优价廉的药品,大大减少了患者的经济负担。本研究的重点就在此。
[Abstract]:Osteoarthritis (osteoarthritis, OA) is a chronic disease, the degeneration of articular cartilage and bone hyperplasia is the common manifestation of some activities, wear, pressure large joints involved earlier incidence of.OA increased year by year, the medical cost of the patients is increasing year after year, especially the elderly joint muscle tissue degradation cause trauma ratio is also more and more high, knee joint OA become the number one killer of elderly disabled, so that OA gradually from medical problems to social problems in the aging society, OA has become key diseases of public health. The Chinese Medical Association orthopedics society established the "guidelines for the diagnosis and treatment of osteoarthritis." the treatment of osteoarthritis of the goal is to reduce joint pain, inhibit the further damage of the joint, as far as possible to keep and improve the joint function, which can greatly improve the quality of life of patients with. Non drug treatments combined with drugs most commonly used drugs include non steroidal anti-inflammatory drugs (NSAIDS), painkillers, viscosupplemetation drugs, glucosamine. Drug therapy can alleviate the symptoms, so the focus of the research is to slow the course of OA to the greatest degree of various biological and biomechanical factors of OA through lots of research target. OA showed that the pathological mechanism of apoptosis cell and cartilage extracellular matrix degradation caused by various reasons caused by joint injury disease treatment targets. Slow or even reverse the apoptosis of cartilage cells and cartilage cells and synovial cells with high expression of matrix metalloproteinase system (MMPs), the enzymes and tissue inhibitor of metalloproteinases-1 (TIMPs) dynamic balance is broken, in the action of mechanical load and inflammatory factor influence, appear in the cartilage matrix free, proteoglycan, soft Persistent bone collagen destruction, resulting in degradation and destruction of cartilage.FDA released the osteoarthritis research guidelines in 1999, including the application of animal model. The OA model is to reproduce the joint disease, development of potential treatments. In this study, selection of New Zealand white rabbits, human reproduction after trauma OA model, the establishment of rabbit osteoarthritis model. Drug therapy for the successful modeling of the animal drug in the experiment. There are two kinds of acetylsalicylic acid and diclofenac sodium. Acetylsalicylic acid is a kind of antipyretic analgesics, can be used for osteoarthritis and rheumatoid disease, can inhibit platelet aggregation. Diclofenac, belongs to non steroidal anti-inflammatory drugs. Since the beginning of 1960s, there are two kinds of agents in the treatment of osteoarthritis and rheumatoid arthritis, but the mechanism of the combined use of drugs and therapeutic targets are not detailed Reported. This study hypothesizes that the pathogenesis of OA, involving MMP, TIMPs, IL-1 beta, NO changes and other factors, intends to OA treatment and the expression of several cytokines in the process of the activity of related research. This study will be divided into the establishment of rabbit osteoarthritis model five.1. Hulth method to the experimental research 2. acetylsalicylic acid combined with diclofenac on rabbit articular cartilage histological effects of 3. acetylsalicylic acid combined with diclofenac on IL-1 rabbit arthritis synovial beta, the effect of NO content of 4. acetylsalicylic acid combined with diclofenac on MMP-3 rabbit articular cartilage cells, MMP-13 expression of 5. acetylsalicylic acid combined with diclofenac on influence the expression of TIMP-1 in rabbit joint cartilage cells: Experimental Study on the effects of different concentrations of acetylsalicylic acid combined with diclofenac on cartilage in rabbit osteoarthritis models. Methods: the combination of acetylsalicylic acid According to the different concentration of diclofenac and divided into three groups: 5mg/kg, 10mg/kg, 20mg/kg, rabbit osteoarthritis model of 32 New Zealand rabbits were established, scoring for each group of rabbits, the determination of NO. content of each IL-1 beta in synovial fluid by using the reagent kit for determination of MMP-3, WB, protein expression of MMP-13, expression of TIMP-1 was measured by PCR method mRNA, to observe the changes of the indexes. Results: the different concentrations of the drug has good effects on treating osteoarthritis in rabbits, can reduce the score, reduce the content of NO, IL-1 beta, WB can see MMP-3, the expression of MMP-13 protein expression, PCR assay showed that the TIMP-1 expression of mRNA. Conclusion: the development of different concentrations of acetylsalicylic acid combined with diclofenac can effectively inhibit rabbit osteoarthritis, and can inhibit NO, MMP-3, MMP-13, IL-1 beta, and increase the expression of TIMP-1, which plays a role in the treatment of OA. Through the treatment process of several fine To detect the expression and activity of cytokines, the molecular mechanism of the combination are discussed, which lays a solid foundation for combination therapy. But the two drugs are cheap drugs, greatly reduce the economic burden of the patients. The focus of this study here.

【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R684.3;R-332

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