三水白虎汤对人肿瘤坏死因子-α转基因关节炎小鼠滑膜组织自噬的影响

Effects of Sanshui Baihu Decoction on the autophagy of synovial tissue in human tumor necrosis factor-a transgenic mice

目的:通过观察三水白虎汤(SSBH)对人肿瘤坏死因子-α转基因关节炎(Tgtc)小鼠滑膜组织自噬的影响,以阐明SSBH减轻关节炎的作用机制。方法:24只雌性Tgtc小鼠按随机数表法分为空白对照(BC)组、SSBH组和阳性对照(3-甲基腺嘌呤,3-MA)组,8只雌性FVB小鼠作为阴性对照(NC)组,药物连续灌胃30d后评估关节变形指数、关节组织病理,透射电镜观察滑膜组织自噬体,ELISA法、免疫组化及免疫荧光检测自噬相关蛋白。结果:SSBH组、3-MA组小鼠关节变形指数、关节病理评分较BC组低(P<0.05);3组Tgtc小鼠滑膜组织中在透射电镜下可观察到自噬体;3组Tgtc小鼠滑膜组织不同程度表达微管相关蛋白1A/1b-轻链3B(LC3B),且SSBH、3-MA组表达量较BC组低(P<0.05);SSBH、3-MA组Tgtc小鼠血清、滑膜组织免疫组化及免疫荧光中雷帕霉素靶蛋白(mTOR)表达量较BC组高(P<0.05),Atg1蛋白激酶复合体(ULK1)较BC组低(P<0.05)。结论:Tgtc小鼠关节滑膜组织的自噬水平升高,SSBH治疗Tgtc小鼠关节炎的作用机制可能为上调自噬通路中的mTOR及下调ULK1。

Objective:To investigate the effects of Sanshui Baihu Decoction(SSBH)on the autophagy of synovial tissue in human tumor necrosis factor-α(TNF-α)transgenic(Tgtc)mice,and to clarify the mechanism of SSBH in reducing arthritis.Methods:Twenty-four female Tgtc mice were randomly divided into blank control(BC)group,SSBH group and positive control(3-methyladenine,3-MA)group.Eight female FVB mice were used as negative control(NC)group.After continuous intragastric administration of drugs for 30 days,joint deformation index and joint histopathology were evaluated.Autophagosomes were observed by transmission electron microscopy,and autophagy-related proteins were detected by ELISA,immunohistochemistry and immunofluorescence.Results:The joint deformation index and pathological score of mice in SSBH group and 3-MA group were lower than those in BC group(P<0.05).Autophagosomes could be observed under transmission electron microscope in synovial tissues of Tgtc mice in three groups.Microtubule-associated protein 1A/1B-light chain 3B(LC3B)was expressed in synovial tissues of Tgtc mice in different degrees,and the expression levels of SSBH group and 3-MA group were lower than those of BC group(P<0.05).The expression of target of rapamycin(mTOR)in serum and synovial tissue of Tgtc mice in SSBH group and 3-MA group was higher than that in BC group(P<0.05).Atgl protein kinase complex(ULKI)was lower than that in BC group(P<0.05).Conclusion:The level of autophagy in the joint synovium of Tgtc mice is increased.The mechanism of SSBH in the treatment of arthritis in Tgtc mice may be up-regulation of mTOR and down-regulation of ULKI in the autophagy pathway.