青蒿琥酯对佐剂性关节炎大鼠血清及滑膜细胞培养上清液中肿瘤坏死因子α及趋化因子的影响

Effects of Artesunate on Tumor Necrosis Factor α and Chemotactic Factors in the Serum and the Synoviocyte Culture Supernate of Collagen-induced Arthritis Rats

目的验证青蒿琥酯对佐剂性关节炎大鼠血清及滑膜细胞原代培养上清液中单核细胞趋化因子(MCP)-1、调节正常T细胞表达和分泌因子(RANTES)和肿瘤坏死因子(TNF)-α的影响。方法取雄性Wistar大鼠80只建立佐剂性关节炎大鼠模型,第6天选出右后足和两前足关节炎指数之和≥6分的60只大鼠,随机数字表法分为空白对照组、模型对照组、甲氨蝶呤阳性药物对照组(简称甲氨蝶呤组)和青蒿琥酯大剂量[20mg/(kg.d)]、中剂量[10mg/(kg.d)]、小剂量组[2.5mg/(kg.d)]并灌胃给药。给药免疫后第8天处死大鼠,抽取静脉血,取血清。同时无菌操作取各组大鼠膝关节滑膜组织行原代培养后培养48h,离心收集上清液。采用酶联免疫吸附试验(ELISA)观察各组治疗前后血清及滑膜细胞原代培养上清液MCP-1、RANTES及TNF-α浓度变化。结果青蒿琥酯大、中、小剂量组能显著降低血清及上清液中细胞因子TNF-α,与模型对照组比较,差异有统计学意义(P<0.05),青蒿琥酯大、中剂量组与甲氨蝶呤组比较,差异无统计学意义(P>0.05),而青蒿琥酯小剂量组与青蒿琥酯大、中剂量组及甲氨蝶呤组比较,差异均有统计学意义(P<0.05);青蒿琥酯各剂量组能显著降低血清及上清液中炎性趋化因子MCP-l和RANTES的表达,与模型组比较,差异有统计学意义(P<0.05),而与甲氨蝶呤组比较,差异无统计学意义(P>0.05)。结论青蒿琥酯抗炎和免疫调节作用可能与抑制炎症因子TNF-α及趋化因子MCP-1、RANTES相关。

Objective To evaluate the effects of Artesunate on tumor necrosis factor α（TNF-α）,monocyte chemoattractant protein（MCP-1）,and on reduced activation normal T cell expressed and secreted（RANTES） in the serum and the synoviocyte culture supernate of collagen-induced arthritis（CIA） rats.Methods Eighty male Wistar rats were selected to establish the CIA rat model.On the 6th day after modeling,60 rats with the sum of arthritis index of right metapedes and two propodium ≥6 were selected,and randomly divided into 6 groups（n=10）,i.e.,the blank control group,the CIA model control group（treated with normal saline,abbreviated as the CIA group）,the MTX positive control group（abbreviated as the MTX group）,the large dose Artesunate group（at the daily dose of 20 mg/kg）,the moderate dose Artesunate group（at the daily dose of 10 mg/kg）,and the small dose of Artesunate group（at the daily dose of 2.5 mg/kg）.Mice were sacrificed 7 days of immune injection and their venous blood was collected to obtain the serum.Meanwhile,the synovial tissues of the knee joint were taken by aseptic techniques and primary cultured for 48 h.The supernate was collected by centrifuge.The changes of MCP-1,RANTES,and TNF-α in the serum and the synoviocyte culture supernate were observed in each group before and after treatment using ELISA.Results Artesunate significantly decreased the expressions of TNF-α in the serum and the synoviocyte culture supernate,showing significant difference when compared with the model control groups（P0.05）.There was no statistical difference in the large dose Artesunate group and the moderate dose Artesunate group when compared with the MTX group（P0.05）.But statistical difference existed in the large dose Artesunate group,the moderate dose Artesunate group,and the MTX group when compared with the small dose Artesunate group（P0.05）.Artesunate could significantly decrease the expressions of MCP-1 and RANTES in the serum and the synoviocyte culture supernate,showing statistical difference when compared with the model control group（P0.05）.But no statistical difference existed when compared with the MTX group（P0.05）.Conclusion The mechanism of anti-inflammatory action and immune regulation of Artesunate might be correlated with the inhibition of inflammatory factor TNF-α and chemotactic factors MCP-1 and RANTES.