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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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)
