甘草次酸对胶原诱导性关节炎滑膜成纤维样细胞炎症细胞因子表达和分泌的影响  

Effect of glycyrrhetinic acid on the expression of inflammatory factors in fibroblast-like synovial cells from collagen induced arthritis rats

作　　者：桂明[1] 李扬[1] 伍婷[1] 李英[1] 黄莹[1] 张浩[1] 

机构地区：[1]中南大学湘雅三医院肾内科,湖南长沙410013

出　　处：《细胞与分子免疫学杂志》2012年第9期915-919,922,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：2009年湖南省中医药科研计划(2009063)

摘　　要：目的:观察甘草次酸(GA)对胶原诱导性关节炎大鼠滑膜成纤维样细胞(CIA-FLS)TNF-α、IL-1β表达分泌的影响,并观测与传统治疗药物MTX联合应用后的效果。方法:组织块培养法体外分离纯化CIA大鼠膝关节滑膜组织中CIA-FLS细胞。实验分为空白组、MTX组、GA组和GA+MTX组4组,分别对CIA-FLS细胞进行体外药物干预实验。每组分别干预1、3、5 d。Real time qPCR法检测各组CIA-FLS细胞TNF-α、IL-1β mRNA表达水平,ELISA法检测细胞培养上清液中TNF-α、IL-1β水平。结果:药物干预1 d,CIA-FLS TNF-α、IL-1β表达均降低,但与对照组比较差异无统计学意义(P>0.05)。药物干预3 d,CIA-FLS TNF-α、IL-1β表达均降低,与对照组比较差异有统计学意义(P<0.05),3组组间差异无统计学意义(P>0.05)。药物干预5 d,CIA-FLS MTX、GA、GA+MTX均可明显抑制TNF-α、IL-1β表达(P<0.05),处理组间差异有统计学意义,按照强弱依次是GA+MTX>MTX>GA(P<0.05)。MTX、GA、GA+MTX抑制TNF-α、IL-1β表达均呈时效依赖性,作用时间延长,抑制作用越明显(P<0.05)。结论:GA、MTX、GA+MTX对CIA-FLS炎症因子TNF-α、IL-1β表达和分泌的抑制作用依次递增,并呈时间依赖性。AIM：To determine the effects of glycyrrhetinic acid（GA）,methotrexate（MTX） and GA＋MTX on the expressions of tumor necrosis factor-α（TNF-α） and interleukin-1β（IL-1β） in fibroblast-like synovial（FLS） cells from collagen induced arthritis（CIA） rats.METHODS： CIA-FLS cells were isolated and purified from the synovium of CIA rats by tissue block culture method,and then treated with MTX,GA,GA＋MTX or nothing（control group）,respectively.Each group was cultured for 3 different time periods（1,3,5 d）.Real-time qPCR was used to detect the expressions of TNF-α and IL-1β mRNA in CIA-FLS cells and ELISA to determine the levels of TNF-α and IL-1β in the supernatant.RESULTS： Day 1： No significant down-regulation of TNF-α and IL-1β mRNA was identified in intervention groups compared with the control group（P〉0.05）;Day 3： Intervention groups significantly down-regulated TNF-α and IL-1β mRNA（P〈0.05 vs control）,but without intergroup difference（P〉0.05）;Day 5： the three intervention groups all suppressed the expressions of TNF-α and IL-1β mRNA（P〈0.05 vs control）,with GA＋MTX〉MTX〉GA in inhibiting effect（P〈0.05）.All intervention groups were found to inhibit the expressions of TNF-α and IL-1β in a time-dependent manner（P〈0.05）.Similar outcomes were observed in the change of the levels of TNF-α and IL-1β in cell culture supernatant by ELISA.CONCLUSION： GA,MTX,GA＋MTX suppress the expression of TNF-α and IL-1β in CIA-FLS cells in a time-dependent manner,and the suppressing effect is GA＋MTX〉MTX〉GA.

关 键 词：类风湿关节炎 胶原诱导性关节炎 甘草次酸 滑膜成纤维样细胞 白介素-1 肿瘤坏死因子Α 

分 类 号：R392.11[医药卫生—免疫学]