【作者】 贾明睿；

【导师】 周翠英；

【作者基本信息】 山东中医药大学 ， 中西医结合临床， 2008， 博士

【摘要】 目的:通过骨痹健方用于兔制动法骨关节炎造模进程的对照研究,评估其用于骨关节炎早期预防的疗效和安全性;观察对制动法兔骨关节炎模型造模过程预先、同步、延后应用骨痹健方后关节功能和关节软骨的相应变化及对基质金属蛋白酶-1的调节作用,评估其用于骨关节炎早期预防的最佳时机,为中医中药“治未病”“治欲病”理论用于预防骨关节炎提供实验依据。在本文中也一并对骨关节炎的中医预防策略展开探讨。方法:1、选用健康新西兰大白兔40只,随机分为四组,以在造模进程中同步使用骨痹健方为一组,以同步使用西药伊索佳(化学名硫酸氨基葡萄糖)为药物对照组,并建立空白对照组和单纯造模对照组,以制动法开始兔骨关节炎造模进程,在30天后观察动物造模关节的临床表现、关节软骨情况和病理变化,并监测各组的血常规、肝肾功能和体重。2、选用健康新西兰大白兔30只,随机分为三组,以在造模进程中同步使用骨痹健方为一组,预先应用骨痹健方为一组,延后应用骨痹健方为一组,以制动法开始兔骨关节炎造模进程,在造模30天后观察动物造模关节的临床表现、关节软骨情况和病理变化,免疫组化方法观察软骨中基质金属蛋白酶-1的表达,并监测各组的血常规、肝肾功能和体重。结果:1、骨痹健方组的临床表现评分明显优于单纯造模组(P<0.05),与硫酸氨基葡萄糖组没有显著性差异(P>0.05);骨痹健方组关节软骨大体情况和光镜下病理变化评价明显优于模型组,与硫酸氨基葡萄糖组没有明显差异;骨痹健方组Mankin’s评分与模型组比较有非常显著性差异(P<0.01),与硫酸氨基葡萄糖组没有显著性差异(P>0.05);各组安全性指标没有明显差异(P>0.05)。空白对照组的临床表现评分、关节软骨大体情况和光镜下病理变化评价及Mankin’s评分与其他三组比较均有显著性差异(P<0.05)。2、预先应用骨痹健方组的临床表现指标中的局部疼痛刺激反应和关节肿胀指标明显优于同步应用骨痹健方组和延后应用骨痹健方组(P<0.05),步态改变和关节活动范围指标明显优于延后应用骨痹健方组,与同步应用骨痹健方组没有显著性差异(P>0.05);同步应用骨痹健方组的各项临床表现指标均明显优于延后应用骨痹健方组(P<0.05);预先应用骨痹健方组软骨大体情况、光镜下病理变化评价、Mankin’s评分明显优于同步应用骨痹健方组,同步应用骨痹健方组软骨大体情况、光镜下病理变化评价、Mankin’s评分明显优于延后应用骨痹健方组(P<0.05);预先应用骨痹健方组的基质金属蛋白酶-1表达率明显低于同步应用骨痹健方组,同步应用骨痹健方组的基质金属蛋白酶-1表达率明显低于延后应用骨痹健方组(P<0.05)。各组安全性指标没有明显差异(P>0.05)。结论:1、骨痹健方能显著降低制动法兔骨关节炎动物模型造模进程导致的软骨病理损伤、改善临床表现和组织学评分,以在造模前预先用药组的疗效最为明显。2、骨痹健方能抑制兔骨关节软骨基质金属蛋白酶-1的活化,调节细胞因子,从而抑制了造模进程中软骨基质的降解,以在造模前预先用药组的调节作用最为明显。3、该方虽不能阻止实验性骨关节炎病理进程的发展,但却能延缓其发展过程,早期用药组的疗效优于较晚用药组,并且没有观察到用药过程中实验兔出现血液指标和肝肾功能的明显损害。更多还原

【Abstract】 Objective:Through the comparison study on the therapy influences, which are brought by Gubijian Formula, in the process of establishing experimental rabbit model of osteoarthritis introduced by the joint immobilization method, its preventive effect of early treatment against osteoarthritis and its safety of medical treatment are evaluated. Through the observations made to changes of joint functions and joint cartilages,and adjustment functions to matrix metalloproteinases-1 , which are corresponding to pre-treated,synch-treated and deferred treatment experimental rabbit groups,during the process of establishing rabbit models of osteoarthritis induced by the joint immobilization method, the evaluation to the best period of the early preventive treatment against osteoarthritis using Gubijian Formula is performed. In the field of osteoarthritis treatment, the thesis provides the experimental basis to the theories of Chinese traditional medicine, which are“To take the preventive treatment is a doctor’s first priority”and“The doctor should pay great attention to people who are in their sub-health conditions”. In this article, the discussion regarding the preventive strategy against osteoarthritis,which originates from the chinese traditional medicine,is also developed.Designs: 1. Forty healthy New Zealand white rabbits are selected and divided to four groups randomly. As soon as the process of establishing experimental rabbit model of osteoarthritis is started,one group is given the treatment of Gubijian Formula and another group is given the treatment of glucosamine sulfate. These two groups are the medicine -treated control groups. Also, the blank control group and the control group that is only for establishing the model of osteoarthritis are organized. The process of establishing the experimental rabbit model of osteoarthritis is started by adopting the joint immobilization method. After 30 days , the observation is made to the clinical performances of joints that are used to establish the model of osteoarthritis,main situations of joint cartilages and pathological changes under the light microscope,and the monitoring is performed to the blood routine examinations,hepatic and renal functions and weights of each control group. 2. Thirty healthy New Zealand white rabbits are selected and divided to three groups randomly. The synch-treated control group is treated with Gubijian Formula as soon as the process of establishing the model of osteoarthritis is started. The pre-treated control group was treated with Gubijian Formula a few days before the process is started. The last group with the deferred treatment will be treated with Gubijian Formula a few days after the process is started. The process of establishing the experimental rabbit model of osteoarthritis is started by adopting the joint immobilization method. After 30 days, the observation is made to the clinical performances of joints that are used to establish the model of osteoarthritis,main situations of joint cartilages and pathological changes under the light microscope,and the monitoring is performed to the blood routine examinations,hepatic and renal functions and weights of each control group. At the same time,with the immunohistochemical method, the observation is made to the expression of matrix metalloproteinase-1 in cartilages.Results: 1. The grade of clinical performance gotten from the control group treated with Gubijian Formula is much better than the control group that is only for establishing the experimental model of osteoarthritis(P<0.05),and does not have significant difference with the group treated with the glucosamine sulfate(P>0.05). The evaluation of main situations of cartilages and pathological changes under the light microscope,which is gotten from the control group with the treatment of Gubijian Formula, is much better than the control group with the pure model establishment of osteoarthritis,and does not have the significant difference with the control group treated by the glucosamine sulfate. Mankin’s Marks that are gotten from the control group with the treatment o f the Gubijian Formula and the control group with the pure model establishment of osteoarthritis have a significant difference(P<0.01),but Mankin’s Marks of two groups with the Gubijian Formula treatment and the glucosamine sulfate treatment do not have a significant difference(P>0.05). There is no significant difference in the safety indexes of four control groups (P>0.05). Comparing to the other three control groups, there are some significant differences in the blank control group in terms of the clinical performance , situations of cartilages and pathological changes under the light microscope and the Mankin’s Mark(P<0.05). 2. The indexes of clinical performance including the indexes of local pain reaction and joint swelling, which is gotten from the pre-treated control group, are much better than the synch-treated and deferred treatment control groups(P<0.05). The indexes of the gait change and motion range of joints, which are gotten from the pre-treated control group, are much better than the deferred treatment group and do not have the significant difference with the synch-treated group(P>0.05).Every index of clinical performance gotten from synch-treated group is much better than the deferred treatment group(p<0.05). The main situations of cartilages,evaluation of pathological changes under the light microscope and Mankin’s Mark,which are gotten from the pre-treated control group,are much better than the synch-treated control group. The main situations of cartilages,evaluation of pathological changes under the light microscope and Mankin’s Mark,which are gotten from the synch-treated control group,are much better than the deferred treatment control group(P<0.05).The expression rate of Matrix Metalloproteinase-1 of the pre-treated control group is much lower than the synch-treated control group(P<0.05). The expression rate of Matrix Metalloproteinase-1 of the synch-treated control group is much lower than the deferred treatment control group(P<0.05).There is no significant difference in the safety indexes of three control groups (P>0.05).Conclusions: 1. The Gubijian Formula can effectively reduce the pathological injury intensity caused by the process of establishing the experimental rabbit model of osteoarthritis introduced by the joint immobilization method,and improve the clinical performance and histological grade. The control group that was treated with The Gubijian Formula before the model establishment of osteoarthritis has the most obvious medical effect. 2. The Gubijian Formula can control the activation of Matrix Metalloproteinase-1 of rabbits’joint cartilages and adjust the cytokines. Therefore,it can inhibit the egradation of the cartilage matrixes during the process of establishing the model of osteoarthritis. The control group that was treated with The Gubijian Formula before the model establishment of osteoarthritis has the most obvious adjustment function. 3. Although the formula can not prevent the pathological development of the experimental osteoarthritis,it can slow down its progress. The control group that is treated early has better therapeutic effect than the control group that is treated late. No obvious adverse influence is found in the blood indexes and hepatic and renal functions of experimental rabbits.更多还原