摘要
目的:探讨白芍总苷(TGP)对白介素22(IL-22)诱导的HaCaT银屑病细胞模型的抗凋亡作用,探索其在银屑病治疗中的机制,为银屑病的临床研究用药提供理论依据.方法:IL-22刺激HaCaT细胞,检测细胞活性,筛选IL-22干预的最佳剂量和最佳作用时间,构建银屑病细胞模型.用逆转录聚合酶链式反应(RT-PCR)和蛋白免疫印迹法(WB)测定银屑病细胞模型中的抗凋亡分子的表达.TGP处理银屑病细胞模型后,用MTT检测细胞的活性.免疫荧光染色测定银屑病细胞模型中STAT3的定位.抑制STAT3的表达后,用RT-PCR和WB检测下游抗凋亡分子BCL-XL的表达.实验采用随机分组,通过GraphPad-Prism 6软件对数据进行处理,结果差异进行t检验.结果:MTT实验发现,IL-22处理HaCaT细胞的最佳剂量和时间分别为12.5 mmol/L,48 h.银屑病细胞模型中,抗凋亡相关通路JAK/STAT3/BCL-XL的表达显著增加.通过TGP处理后,细胞活性明显下降,同时细胞核内STAT3明显减少.抑制STAT3表达后,银屑病细胞模型活性指标显著降低,尤其是STAT3下游的分子BCL-XL的表达下降.结论:TGP基于调节抗凋亡通路JAK/STAT3/BCL-XL的表达,降低IL-22诱导的银屑病细胞模型的活性.
Abstract
Objective:IL-22 was applied to stir the HaCaT cells to construct psoriasis model and to detect the changes of anti-apoptotic signaling molecule in the model.So in this study,total glucosides of paeony(TGP)was used to treat the cell model to explore the possible therapeutic mechanism for psoriasis.Methods:After HaCaT cells were stimulated with different concentrations of IL-22 for different periods of time,the changes of cell activity were detec-ted by MTT,and the optimal parameters of IL-22 were determined.RT-PCR and WB were used to detect the expres-sion of inflammatory and apoptosis-related molecules in IL-22 induced psoriatic cell models.The MTT and the Im-munofluorescence staining were applied to measure cell activity of psoriasis cell model after treated with TGP.RT-PCR and WB were used to detect the expression of downstream anti-apoptotic molecule BCL-XL after inhibition of STAT3 expression.The experiment was randomly grouped,and Graph Pad-Prism 6 software was used to analyze the results by t test.Results:MTT results showed that the optimal concentration of IL-22 in HaCaT cells was 12.5 mmol/L and the optimal treatment time was 48 h.The expression levels of inflammatory and apoptosis-related molecular signaling pathway J AK/STAT3/BCL-XL were significantly increased in IL-22-induced psoriasis cell model.The activity of Psoriatic cell model was decreased after treated with TGP,as same the localization of STAT3 in nucleus.Inhibition of STAT3 expression significantly reduced the activity of psoriatic cell model and the expression of STAT3 and its downstream anti-apoptotic molecule BCL-XL.Conclusion:TGP could reduce the activity of IL-22 induced psoriasis cell model by inhibiting the expression and activation of inflammatory and apoptotic signaling cascade JAK/STAT3/BCL-XL.
维普
万方数据