加味四妙勇安汤对CIA大鼠关节炎性病理损伤的作用研究

Study on pathological and molecular mechanism of the effect of the modified Simiao Yong'an decoction on the arthritis in CIA rats

关键词:  加味四妙勇安汤  CIA模型  AI  病理改变  分子机制

Key words:  modified Simiao Yong'an decoction  CIA model  AI  pathological change  molecular mechanism

摘要:

[目的]探讨加味四妙勇安汤对Ⅱ型胶原诱导性关节炎（CIA）大鼠关节炎指数（AI）、关节病理损伤的改善情况，对大鼠体内白介素（IL）-6，IL-17，肿瘤坏死因子-α（TNF-α）表达的影响。[方法]建立CIA模型，将造模成功的SD大鼠随机分为模型组、阳性药（来氟米特）组、中药高、中、低剂量组，设正常对照组，每组6只，干预治疗12周。每周评价每只鼠的AI值。12周后麻醉处死，取各组大鼠右踝关节，光镜下观察病理学变化。采用逆转录-聚合酶链反应（RT-PCR）法检测大鼠膝关节滑膜IL-6、IL-17、TNF-α的mRNA转录水平，酶联免疫吸附实验（ELISA）法检测血清IL-6、IL-17、TNF-α的蛋白表达水平。[结果]中药3种剂量组在6~12周时AI分值较模型组明显下降（P<0.05或P<0.01）；而且在8~10周还分别表现出较阳性组显著的改善（P<0.05或P<0.01）。光镜检测结果显示：中药中剂量组明显改善了CIA模型大鼠关节滑膜增生、炎性细胞浸润情况。细胞因子检测结果显示各治疗组IL-6、IL-17、TNF-α的mRNA及蛋白表达显著降低（P<0.05或P<0.01）。[结论]加味四妙勇安汤对CIA模型大鼠关节炎改善作用的分子机制，可能与下调炎性细胞因子IL-6、IL-17、TNF-α的表达相关。

Abstract:

[Objective] To explore the effect of modified Simiao Yong'an decoction on collagen induced arthritis about arthritis index (AI), the joint tissue inflammatory pathological manifestations, and effect on protein expression of IL-6,IL-17,TNF-α in rats.[Methods] The successful CIA models of SD rats were randomly divided into model group, positive group (leflunomide), and high, medium, low dose group of Chinese medicine, and normal control group, 6 rats in each group. The intervention and treatment period was 12 weeks. AI values of each rat were evaluated every week. At 12 weeks, rats were executed and right ankle joint of each group of rats was taken. Joint pathlological changes of each right ankle were observed. At the same time levels of IL-6, IL-17 and TNF-α in serum were examined by ELISA, mRNA transcription levels of IL-6, IL-17and TNF-α in synovial were detected by RT-PCR.[Results] AI scores of the three Chinese medicine groups decreased significantly (P<0.05 or P<0.01) in 6~12 weeks compared with the model group, and in the 8~10 weeks AI scores were even improved significantly than positive group (P<0.05 or P<0.01). Histological examination showed that the ankle joint cartilage pathological had an improvement in the medium dose of modified Simiao Yong'an decoction group compared with the control model group. Compared with the model group, mRNA and protein expression of IL-6, IL-17and TNF-α were down regulated in all treatment groups significantly (P<0.01).[Conclusion] The anti-inflammatory action mechanism on CIA rats of modified Simiao Yong'an decoction might achieve by reducing expression of IL-6, IL-17 and TNF-α.