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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。
Objective: To observe the effect of glycyrrhetinic acid (GA) on the expression and secretion of TNF-α and IL-1β in synovial fibroblastoid cells (CIA-FLS) of rats with collagen-induced arthritis and observe the effect of glycyrrhetinic acid Effect. Methods: CIA-FLS cells were isolated and purified from synovial tissue of knee joint of CIA rats by tissue culture method. The experiment was divided into 4 groups: blank group, MTX group, GA group and GA + MTX group, respectively, in vitro drug intervention experiments on CIA-FLS cells. Each group were interventions 1,3,5 d. The expression of TNF-α and IL-1β mRNA in CIA-FLS cells in each group was detected by Real time qPCR method. The levels of TNF-α and IL-1β in cell culture supernatants were detected by ELISA. Results: The levels of TNF-α and IL-1β in CIA-FLS decreased after 1 d of drug intervention, but there was no significant difference between the two groups (P> 0.05). The levels of TNF-α and IL-1β in CIA-FLS were decreased 3 d after drug intervention, with statistical significance (P <0.05). There was no significant difference between the three groups (P> 0.05). MTX, GA and MTX could significantly inhibit the expression of TNF-α and IL-1β (P <0.05) after 5-day drug intervention. The difference between the two groups was statistically significant, followed by GA + MTX > MTX> GA (P <0.05). MTX, GA, MTX and MTX inhibited the expression of TNF-α and IL-1β in a time-dependent manner, and prolonged the inhibitory effect (P <0.05). CONCLUSION: The inhibitory effects of GA, MTX, GA + MTX on the expression and secretion of TNF-α and IL-1β in CIA-FLS are increasing in a time-dependent manner.