天山雪莲口服液对类风湿关节炎的治疗作用及其机制研究

Therapeutic effectiveness of Sasussured Involucrata Oral Liquid on rheumatoid arthritis and its underling mechanism

目的:观察天山雪莲口服液(SIOL)对类风湿关节炎(RA)的治疗作用,并探讨其作用机制。方法:将50只雄性SD大鼠随机分为健康对照组、阴性对照组、阳性对照组及SIOL低、高剂量组,10只/组。通过注射牛Ⅱ型胶原建立胶原诱导大鼠关节炎模型(CIA),健康对照组和阴性对照组灌胃生理盐水(2 ml/d),阳性对照组灌胃雷公藤多苷片[20 mg/(kg·d)],SIOL低、高剂量组分别灌胃SIOL(0.5 ml/d、2.0 ml/d),共给药28 d。给药期间每周测量并计算大鼠左后踝关节肿胀度;造模给药后采用HE染色观察大鼠关节滑膜组织病理形态表现;qPCR检测关节滑膜组织JNK、Bcl-2、Beclin-1、LC3 mRNA表达水平;Western blot检测JNK、p-JNK、Bcl-2、Beclin-1、LC3-Ⅱ相对蛋白表达水平。结果:与健康对照组相比,阴性对照组大鼠关节肿胀度明显加重(P<0.01),关节滑膜组织中JNK、Bcl-2、Beclin-1、LC3 mRNA表达均显著升高(P<0.01),关节滑膜组织中p-JNK、Bcl-2、Beclin-1、LC3-Ⅱ相对蛋白表达水平显著升高(P<0.05,P<0.01)。与阴性对照组相比,阳性对照组及SIOL各剂量组大鼠关节肿胀度明显减轻(P<0.01),关节滑膜组织中JNK、Bcl-2、Beclin-1、LC3 mRNA表达显著降低(P<0.05,P<0.01),关节滑膜组织中p-JNK、Bcl-2、Beclin-1、LC3-Ⅱ相对蛋白表达水平显著降低(P<0.05,P<0.01)。结论:SIOL对CIA大鼠具有治疗作用,其机制可能与抑制滑膜组织JNK/Bcl-2/Beclin-1介导的自噬通路激活有关。

Objective:To observe therapeutic effects of Saussurea Involucrata Oral Solution(SIOL)on rheumatoid arthritis and investigate its mechanisms of action.Methods:Fifty male SD rats were randomly divided into healthy control,negative control,positive control,low-dose SIOL and high-dose SIOL group(10 rats per group). Rat model of collagen-induced arthritis(CIA)was then established through injection of rodents with bovine type Ⅱ collagen. Over a period of 28 days,rats in healthy control and negative control groups were administered saline(2 ml/d)by gavage,those in positive control group were administered tripterygium glycoside tablets[20 mg/(kg·d)]by gavage,and those in low-dose and high-dose SIOL groups were administered SIOL(0.5 ml/d,2.0 ml/d)by gavage. During the drug administration period,degree of swelling in left hind ankle joint of each rat was measured and calculated once a week. After CIA model construction and drug administration,synovial tissue was dissected from rats and subjected to HE staining for observation of pathomorphological characteristics,qPCR analysis for measurement of JNK,Bcl-2,Beclin-1 and LC3 mRNA expression levels,and Western blot assay for measurement of relative JNK,p-JNK,Bcl-2,Beclin-1 and LC3-Ⅱ protein expression levels.Results:Compared with healthy control group,negative control group exhibited significant increases in degree of joint swelling(P<0.01),levels of JNK,Bcl-2,Beclin-1 and LC3 mRNA in synovial tissue(P<0.01)and relative expression levels of p-JNK,Bcl-2,Beclin-1 and LC3-Ⅱ protein in synovial tissue(P<0.05,P<0.01). Compared with negative control group,positive control group and both low-dose and high-dose SIOL groups showed significant decreases in degree of joint swelling(P<0.01)and in levels of JNK,Bcl-2,Beclin-1 and LC3 mRNA in synovial tissue(P<0.05,P<0.01)and relative expression levels of p-JNK,Bcl-2,Beclin-1 and LC3-Ⅱprotein in synovial tissue(P<0.05,P<0.01).Conclusion:SIOL provided therapeutic effects in rats with CIA,its mechanisms of action may be related to inhibition of JNK/Bcl-2/Beclin-1-mediated autophagy pathway activation in synovial tissue.