白芍总苷对类风湿关节炎成纤维样滑膜细胞增殖和凋亡的影响及机制

Effects and Mechanism of Total Glucosides of Paeony on the Proliferation and Apoptosis of Fibroblast-Like Synovial Cells in Rheumatoid Arthritis

目的:探究白芍总苷(TGP)对类风湿关节炎成纤维样滑膜细胞增殖、凋亡以及炎症反应的影响及其作用机制。方法:将人类风湿关节炎成纤维样滑膜细胞(HFLS-RA)分为对照组、模型组和干预组,其中对照组细胞常规培养;模型组和干预组细胞使用肿瘤坏死因子(TNF)-α10 ng/mL培养建立RA体外细胞模型;随后干预组细胞使用TGP 62.5μg/mL培养,对照组和模型组细胞使用等量生理盐水培养;采用CCK-8法检测细胞增殖能力;原位末端转移酶标记技术(TUNEL)检测细胞的凋亡率;酶联免疫吸附试验法(ELISA)检测白细胞介素(IL)-6和血管内皮生长因子(VEGF)水平;Western Blotting检测磷酸化JAK2蛋白(p-JAK2)、磷酸化STAT1蛋白(p-STAT1)以及磷酸化STAT3蛋白(p-STAT3)的表达水平。结果:CCK-8和TUNEL结果显示TGP可抑制HFLS-RA细胞增殖,促进细胞凋亡;ELISA结果显示TGP可降低细胞中IL-6和VEGF的水平;Western Blotting实验显示TGP可降低p-JAK2、p-STAT1和p-STAT3的表达水平(P<0.05)。结论:TGP可通过调节JAK/STAT信号通路抑制HFLS-RA细胞增殖,促进细胞凋亡,抑制滑膜细胞炎症反应和血管生成。

Objective:To investigate the effects of total glucosides of paeony(TGP)on the proliferation,apoptosis and inflammatory response of fibroblast synovial cells in rheumatoid arthritis and its mechanism.Methods:Human fibroblast-like synoviocytes(HFLS-RA)were divided into a control group,a model group and an intervention group randomly,and the cells of the control group were cultured routinely.Cells in the model group and intervention group were cultured with tumor necrosis factor(TNF)-α10 ng/mL to establish RA cell models in vitro.Subsequently,cells in the intervention group were cultured with TGP 62.5μg/mL,and cells in the control group and model group were cultured with the same amount of normal saline.CCK-8 assay method was used to detect cell proliferation.Cell apoptosis rate was detected by in situ end transferase labeling(TUNEL).The levels of interleukin(IL)-6 and vascular endothelial growth factor(VEGF)were determined by enzyme-linked immunosorbent assay(ELISA).The expression levels of phosphorylated JAK2(p-JAK2),phosphorylated STAT1(p-STAT1)and phosphorylated STAT3(p-STAT3)were detected by Western blotting.Results:CCK-8 and TUNEL assay showed that TGP could inhibit the proliferation of HFLS-RA cells and promote cell apoptosis.ELISA assay showed that TGP can reduce the levels of IL-6 and VEGF in cells.Western blotting assay showed that TGP can reduce the expression levels of p-JAK2,p-STAT1 and p-STAT3(P<0.05).Conclusion:TGP can inhibit HFLS-RA cell proliferation,promote cell apoptosis,inhibit synovial cell inflammation and angiogenesis by regulating JAK/STAT signaling pathway.