关节镜联合电针治疗膝骨性关节炎的分子机制研究

The Study of Molecular Mechanism on Electro-acupuncture combined with Arthroscope for Knee Osteoarthritis

目的：研究关节镜清理术联合电针治疗膝骨关节炎的分子机制,通过观察关节液中影响膝关节软骨生理病理过程的MMP13、IL-1、TNF-a、TGF-β细胞因子,探讨中西医结合治疗膝骨关节炎的临床疗效机制。方法：治疗组和对照组术前、术后4周、6个月、12个月均采用免疫组织化学染色方法检测MMP13的mRNA原位表达及采用酶联免疫吸附测定（ELISA）法检测各组关节液中IL-1、TNF-α、TGF-β的含量,对患者治疗前后关节液中细胞因子和蛋白酶进行量化比较。结果：1术前两组患者的IL-1、TNF-α、TGF-β及MMP13的比较差异无统计学意义（P〉0.05）;2治疗组4周时IL-1、TNF-α含量较术前均有显著下降（P〈0.05）,6个月时TGF-β含量较术前显著升高,对照组较治疗组同时期的IL-1、TNF-α、TGF-β的改变滞后;3两组术后6个月时IL-1、TNF-α指标显著改变,而TGF-β的含量变化较IL-1、TNF-α改变幅度小;4治疗组在术前与术后4周,术后4周与6个月、术后6个月与术后1年的IL-1、TNF-α比较差异均有统计学意义（P〈0.05）,细胞因子IL-1、TNF-α均有显著下降;5治疗组与对照组观察并不是所有标本都检测到MMP13且MMP13指标变化不明显（P〉0.05）。结论：1无论治疗组还是对照组均可以通过调节IL-1、TNF-α与TGF-β细胞因子生成,并维持分解与生成代谢的动态平衡,达到治疗疗效;2两组对于改善膝骨关节炎疼痛及功能在关节液的表达上,是以降低IL-1、TNF-α对于软骨的破坏为主,而TGF-β对于软骨的修复出现较晚;3对照组于术后6个月关节疼痛才开始逐渐缓解,细胞因子IL-1、TNF-α、TGF-β的改变滞后;42组在术后4周时细胞因子IL-1、TNF-α含量较术前均有显著下降,在术后6个月时细胞因子TGF-β含量较术前显著升高,临床表现为术后4周即关节疼痛开始缓解,在术后6个月时疼痛缓解显著,在术后6个月~1年时间段内关节功能康复疗效显著提高;52组MMP13表达都存在时相特异性的特点,不能验证治疗效果。

Objective：To study the molecular mechanism of electro-acupuncture combined with arthroscope in treating knee osteoarthritis.By observing the Effects of physiological and pathological processes of knee cartilage of MMP13、IL-1、TNF-a、TGF-βin Joint fluid,to discuss the method for the knee osteoarthritis in Integrative Medicine.Methods：The treatment group was treated by electro-acupuncture postoperatively.Immunohistochemical staining was used to detect MMP13 mRNA and enzyme-linked immunoabsorbent assay was used to detect IL-1,TNF-αand TGF-βin the joint fluid before the operation as well as 4weeks,6months and12 months after the operation.Results：1 There were no significant differences in the levels of IL-1,TNF-α,TGF-βand MMP13 between the two groups before the operation（P〉0.05）.2 The content of IL-1and TNF-αin the treatment group was reduced in the first 4weeks after surgery,and TGF-βwas increased significantly 6months after the surgery.At the same time,the control group showed the same trend of changes in IL-1,TNF-αand TGF-β,which were slower.3 There were great changes in IL-1and TNF-αin the two groups,but the change of TGF-βwas not so significant.4IL-1and TNF-αlevels were reduced significantly after the surgery.There was remarkable difference in the levels of IL-1and TNF-αbetween before and after the operation,between4 weeks and 6months after the operation,and also between 6months and 1year after the operation（P〈0.05）.5 MMP13 was not detected in all the two groups,which was not changed significantly（P〉0.05）.Conclusion：1 Either the treatment group or the control group can treat the knee osteoarthritis by adjusting the generated of IL-1,TNF-αand TGF-β,and keep a balance between them.2 The functional improvement in the two groups was mainly through reducing IL-1and TNF-α,leading to less cartilage destruction.TGF-βhad a later function in cartilage repairing.3 The pain in the control group was relieved 6months after the operation,while IL-1,TNF-αand TGF-βwere changed slowly.4 Compared with the control group,postoperative IL-1and TNF-αin the treatment group were reduced significantly（P〈0.05）than before the operation.TGF-βwas increased significantly 6months after the surgery.The pain began to be relieved 4weeks after the operation,and relieved significantly 6months later.Joint function rehabilitation effects were increased significantly between 6months and 12 months after the operation.5 MMP13 level cannot be used to evaluate the therapeutic efficacy either in the treatment group or the control group.