补肾法与活血法对SD大鼠膝骨性关节炎滑膜IL-1β、TNF-α及软骨MMP-13、ADAMTS-5的影响

目的：探讨补肾活血法、活血法、补肾法、祛风法对SD大鼠膝骨性关节炎IL-1β、TNF-α及MMP-13、ADAMTS-5的作用及其机制。方法：取SD大鼠180只,随机分为活血组、祛风组、补肾组、补肾活血组、预防组及模型组,每组各30只,以4%木瓜蛋白酶行膝关节注射制作大鼠膝骨性关节炎模型。模型制作成功后连续给药,预防组在模型制作同时予以活血药。治疗以1个月为1疗程,6组大鼠分别于模型制作成功给药后30、60、90d分批处死。处死后取大鼠膝关节滑膜及软骨,分别运用免疫组化及反转录-多聚酶链反应（RT-PCR）测定SD大鼠膝关节滑膜IL-1β、TNF-α及膝关节软骨组织中MMP-13、ADAMTS-5mRNA含量的表达。结果：模型组与祛风组IL-1β、TNF-α及MMP-13、ADAMTS-5mRNA含量表达显著高于其他各组,且随着治疗进行降低不显著。早期,预防组比其他组降低显著（P〈0.05）;活血组与补肾活血组比较差异无统计学意义（P〉0.05）,但两者比祛风组、补肾组及模型组降低显著（P〈0.05）;晚期补肾活血组、补肾组与其他组比较降低显著（P〈0.05）,且补肾活血组与补肾组比较差异有统计学意义（P〈0.05）。结论：补肾法与活血法对SD大鼠膝骨性关节炎IL-1β、TNF-α及MMP-13、ADAMTS-5mRNA的影响在不同时期具有不同的作用,早期时活血法作用较补肾法显著,且预防性给予活血药治疗效果更佳;晚期补肾法较活血法显著,同时补肾活血法治疗效果最为明显。

Effect of Bushen and Huoxue Method on MMP-13 and ADAMTS-5 in Synovial Membrane and Cartilage of SD Rats with Knee Osteoarthritis

Objective:To detect the effects of Bushenhuoxue,Huoxue,Bushen,and Qufeng method on the expression of IL-1β,TNF-α,MMP-13,and ADAMTS-5 in SD rats with knee osteoarthritis.Methods: All 180 SD rats were randomly divided into 6 groups: Huoxue group,Qufeng group,Bushen group,Bushenhuoxue group,preventive group,and model group,with 30 rats in each group.The model of knee osteoarthritis was made by injecting 4% papain in the knee joint.The rats were continuous administration after modeling successfully,one month for a treatment course.Ten rats in each group were killed and the synovium and knee cartilage were taken in 30,60,and 90 days respectively after treatment.The levels of IL-1β and TNF-α in synovium were detected by immunohistochemistry,while,the expression levels of MMP-13 and ADAMTS-5 mRNA in cartilage were detected by real-time quantitative PCR.Results:The levels of IL-1β,TNF-α and MMP-13,ADAMTS-5 mRNA in the model group and Qufeng group were significantly higher than the other groups,but the lower was not obvious as the treatment progresses.In the early stage,the levels were lowest in the preventive group(P<0.05).There was no difference between Huoxue group and BushenHuoxue group,but they were lower than the other groups except preventive group(P<0.05).In the later stage,the levels in Bushenhuoxue group and Bushen group were decreased significantly than other groups(P<0.05),and there was significant difference between Bushenhuoxue group and Bushen group(P<0.05).Conclusion:Bushen and Huoxue method plays different role in the expression of IL-1β,TNF-α and MMP-13,ADAMTS-5 mRNA in rats with knee osteoarthritis at different stage.In the early stage,the Huoxue method plays an important role.In the later stage,the Bushen method is more effective than the Huoxue method,and the Bushenhuoxue method is the most effective treatment.