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
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R593.22

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

    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 mRNA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 mRNA和蛋白水平降低,Bcl-2 mRNA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 mRNA和蛋白水平升高,Bcl-2 mRNA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。

    Abstract:

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

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高晓鹏,鲁贵生.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[J].新中医,2018,50(4):1-5

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  • 在线发布日期: 2018-04-10
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