五味温通除痹胶囊对佐剂性关节炎大鼠自噬蛋白Beclin-1、LC3-Ⅱ表达的影响

Effects of Wuwei Wentong Chubi Capsules on autophagy proteins of Beclin-1 and LC3-Ⅱ in adjuvant-induced arthritis rats

目的明确五味温通除痹胶囊(桂枝、茯苓、淫羊藿,等)对佐剂性关节炎大鼠自噬蛋白Beclin-1、LC3-Ⅱ表达的影响,探讨其防治类风湿关节炎的可能机制。方法雄性SD大鼠,随机分为正常组、模型组、五味温通除痹胶囊(0.80、1.60、3.20 g/kg)组、阳性药雷公藤多苷片(40 mg/kg)组,每组10只。除正常组外,采用弗氏完全佐剂诱导佐剂性关节炎大鼠模型。致炎后第12天灌胃给药,连续12 d。实验结束后,取非致炎侧踝关节,HE染色观察病理学改变,实时定量荧光PCR测定滑膜组织中Beclin-1、LC3-ⅡmRNA的表达,免疫荧光组织化学染色和Western blot法检测滑膜组织中Beclin-1、LC3-Ⅱ蛋白的表达。结果与模型组相比,五味温通除痹胶囊(1.60、3.20 g/kg)不仅可减轻佐剂性关节炎大鼠关节病理损伤程度,还可显著升高Beclin-1、LC3-ⅡmRNA和蛋白的表达水平。结论五味温通除痹胶囊对佐剂性关节炎大鼠具有一定的治疗作用,其机制可能与促进细胞自噬,减少滑膜细胞的过度增殖,从而减轻关节软骨损伤有关。

AIM To observe the effects of Wuwei Wentong Chubi Capsules（ Cinnamomi Ramulus,Poria,Epimedii Folium,etc.） on autophagy proteins of Beclin-1 and LC3-Ⅱin adjuvant-induced arthritis rats and to explore the possible mechanism of action. METHODS Sixty SD rats were randomized into six groups： normal group,model group,Wuwei Wentong Chubi Capsules（ 0. 8,1. 6,3. 2 g/kg） groups and tripterygium glycosides（ TPT,40 mg/kg） group. In addition to the normal group,adjuvant arthritis（ AA） was induced with Freund＇s complete adjuvant. From the 2th day after injection of FCA,Wuwei Wentong Chubi Capsules with different doses and TPT were given by gavage once a day for 12 days. At the end of the experiment,ankle-joint samples were taken to examine the degree of AA by HE. Beclin-1 and LC3-Ⅱ mRNA were determined by real-time fluorescent quantitative PCR. Meanwhile,the protein expression levels of Beclin-1 and LC3-Ⅱ were determined by immunofluorescence histochemical staining and Western blot. RESULTS As compared with the model group,Wuwei Wentong Chubi Capsules（ 1. 60,3. 20 g/kg） not only significantly reduced histopathological injuries,but also effectively up-regulated mRNA and protein expressions of Beclin-1 and LC3-Ⅱ. CONCLUSION Wuwei Wentong Chubi Capsules has a therapeutic action on AA in rats,which might be partly associated with promoting autophagy,decreasing excessive proliferation of synovial cells,leading to the reduction of damage to articular cartilage.