人参皂苷Rg1抑制滑膜细胞样成纤维细胞侵袭增殖及炎性因子释放的实验研究

Experimental Study of Ginsenoside Rg1 Inhibits the Invasion and Proliferation of Fibroblast-Like Synoviocytes and the Release of Inflammatory Factors

目的:探讨人参皂苷Rg1对滑膜细胞样成纤维细胞(FLSs)增殖、侵袭和炎症因子分泌的影响,为临床治疗类风湿性关节炎(RA)提供理论及实验依据。方法:体外分离培养RA的FLSs。以肿瘤坏死因子-α(TNF-α)刺激FLSs,在上述条件中加入不同浓度Rg1(0,0.4,4及40 mmol/L),培养24 h后,分别用实时荧光定量PCR(RT-PCR),免疫印迹实验(Western blot)和酶联免疫吸附试验(ELISA)检测白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)的表达变化。CCK-8克隆形成以及Transwell评估细胞增殖和侵袭能力。Western-blot评估Rg1对丝裂原活化蛋白激酶/细胞外调节蛋白激酶(MAPK/ERK)通路激活的影响。结果:在基因和蛋白水平,Rg1能抑制由TNF-α诱导FLSs的IL-6和IL-1β的表达,并且限制二者的分泌;此外,Rg1能够显著抑制FLSs的侵袭与增殖;机制上,Rg1抑制了ERK的磷酸化,阻碍了MAPK/ERK通路的激活。结论:Rg1抑制FLSs中MAPK/ERK通路的激活,限制细胞的增殖和侵袭,并阻止炎性因子IL-6和IL-1β的释放,从而在RA的治疗过程中发挥作用。

Objective:To investigate the effects of Ginsenoside Rg1 on the proliferation,invasion and secretion of inflammatory factors of fibroblast-like synoviocytes(FLSs),so as to provide theoretical and experimental basis for the clinical treatment of rheumatoid arthritis(RA).Methods:FLSs of RA were isolated and cultured in vitro.FLSs were stimulated by tumor necrosis factor-α(TNF-α).Different concentrations of Rg1(0,0.4,4 and 40 mmol/L)were added in the above conditions.After 24 hours of culture,expression changes of interleukin-6(IL-6)and interleukin-1β(IL-1β)were detected by real-time fluorescence quantitative PCR(RT-PCR),Western blot and Enzyme-linked immunosorbent assay(ELISA).CCK-8clone formation and Transwell evaluation of cell proliferation and invasion.The effect of Rg1 on the activation of mitogen activated protein kinase/extracellular regulated protein kinase(MAPK/ERK)pathway was evaluated by Western blot.Results:At the gene and protein levels,Rg1 could inhibit the expressions of IL-1βand IL-6 induced by TNF-α,and limit their secretion.In addition,Rg1 can significantly inhibit the invasion and proliferation of FLSs.In mechanism,Rg1 inhibited the phosphorylation of ERK and hindered the activation of MAPK/ERK pathway.Conclusion:Rg1 inhibits the activation of MAPK/ERK pathway in FLSs,limits cell proliferation and invasion,and prevents inflammatory factors IL-6 and IL-1β.And thus play a role in the treatment of RA.