独活寄生汤含药血清对佐剂性关节炎大鼠滑膜成纤维细胞增殖和凋亡的影响

Medicated Serum of Duhuo Jisheng Tang Has Effect on Proliferation and Apoptosis of Fibroblast-like synoviocytes in Rats with Adjuvant-incluced Arthritis

目的:探讨独活寄生汤含药血清对佐剂性关节炎大鼠滑膜成纤维细胞(FLS)增殖和凋亡的影响。方法:采用SD大鼠制备独活寄生汤含药血清,弗氏完全佐剂诱导大鼠佐剂性关节炎,分离提取原代滑膜成纤维细胞。滑膜成纤维细胞分为6组,分别是空白组、模型组、甲氨喋呤组、独活寄生汤高、中、低剂量组,每组5个复孔,分别加入相应的药物。CCK8法检测各组细胞增殖率;荧光定量PCR和ELISA分别检测细胞Bax、Bcl-2、caspase3 m RNA和蛋白水平的表达变化。结果:空白组细胞在各时间点未见明显的变化;模型组细胞随时间延长,增殖抑制率升高(P<0.05);甲氨喋呤组和独活寄生汤高、中、低剂量组细胞随干预时间的延长,增殖抑制率降低(P<0.05)。与空白组比较,模型组细胞在各时间点的增殖抑制率均升高(P<0.05);与模型组比较,甲氨喋呤组和独活寄生汤各剂量组细胞在各时间点的增殖抑制率均降低(P<0.05),尤以高剂量组的抑制率更为明显。与空白组比较,模型组细胞Bax、caspase3 m RNA和蛋白水平降低,Bcl-2 m RNA和蛋白水平升高(P<0.05,P<0.01)。与模型组比较,甲氨喋呤组和独活寄生汤中、高剂量组细胞Bax、caspase3 m RNA和蛋白水平升高,Bcl-2 m RNA和蛋白水平降低(P<0.05,P<0.01),高剂量组的作用更为明显。结论:独活寄生汤能抑制佐剂性关节炎大鼠滑膜成纤维细胞的增殖,其作用机制与下调抗凋亡蛋白Bcl-2,上调促凋亡蛋白Bax和caspase3的表达有关。

Objective：To discuss the effect with of medicated serum of Duhuo Jisheng tang rats on were proliferation applied and apoptosis of Methods：SD to fibroblast-like synoviocytes（FLS）tang.in rats adjuvant-induced arthritis.to produce medicated rats,and then serum of Duhuo Jisheng Complete Freund＇s adjuvant were was used induce adjuvant-induced arthritis in groups：blank group,model group,methotrexate primary FLS were well separated and extracted.tang FLS divided into six group,as low,middle,and dose,with as Duhuo Jisheng groups of high of five replications in each group.Each was group was given corresponding medication respectively.The effect Duhuo Jisheng tang on cell proliferation rate cell Bax,Bcl-2 and caspase3 mRNA as well as detected protein by were the cell counting kit 8（CCK8）and expression changes of the levels and of respectively blank detected by time fluorescence point.quantitative PCR ELISA.Results：There was no obvious change in as the the cells of group at each The The proliferation inhibition rate of cells in the group model group was increased time prolonged（P0.05）.high proliferation was inhibition rate of cells in the time methotrexate and Duhuo Jisheng with tang groups of low,middle,and group,dose decreased cells as the time intervention point prolonged（P0.05）.Compared the blank the proliferation inhibition rate of at each was increased in the model group（P0.05）.Compared with the group,the model proliferation inhibition dose rate of cells at each time the point was decreased in the methotrexate Jisheng group group and Duhuo0.05）,and Jisheng was tang groups of each（Pespecially decrease of rate in Duhuo tang of high group dose were group,levels protein of Bax,caspase3 more obvious.Compared with the blank mRNA and protein in the with model decreased,while 0.05,Pgroup,levels of Bcl-2 mRNA and were increased（P0.01）.Compared the model of Bax,caspase3 mRNA levels were and Bclprotein in methotrexate and group and Duhuo Jisheng tang groups of middle and high dose increased,while 0.05,P 0.01）,and tang levels of 2 mRNA protein obvious.were decreased（Pespecially the effect in in Conclusion：Duhuo Duhuo Jisheng tang group of high dose was more Jisheng can inhibit proliferation of FLS rats with adjuvant arthritis.The mechanism is related to down-regulation and of the expression of anti-apoptotic protein Bcl-2 and up-regulation of the expression of pro-apoptotic protein Bax caspase3.