雷公藤内酯醇抑制小鼠肾小球系膜细胞表达CXCL10的实验研究

摘要

目的：探讨雷公藤内酯醇（triptolide，TPL）对γ干扰素（interferon-γ，IFN-γ）诱导的小鼠肾小球系膜细胞（SV40MES13）表达趋化因子CXCL10 mRNA的影响及其可能机制。方法将对数生长期SV40MES13随机分为空白组、刺激组、干预组，用不同浓度的IFN-γ（0U/ml～2000U/ml）刺激细胞不同时间（0h～48h）后，观察细胞表达CXCL10 mRNA的变化；用CCK-8法检测不同浓度（2ng/ml～20ng/ml）TPL对SV40MES13生存率的影响，选用细胞生存率大于90%的TPL用于后续实验，观察TPL对SV40MES13表达CXCL10 mRNA的影响；选用JAK/STAT信号通路特异性抑制剂AG490干预细胞，探讨IFN-γ是否通过JAK/STAT信号通路诱导CXCL10表达；收集以上细胞，用Real-Time PCR技术检测各组CXCL10 mRNA表达水平。结果 IFN-γ能诱导SV40MES13表达CXCL10 mRNA，并呈时间、剂量依赖性；AG490具有抑制IFN-γ诱导的SV40MES13表达CXCL10 mRNA；TPL具有抑制IFN-γ诱导的SV40MES13表达CXCL10 mRNA的作用。结论 IFN-γ可能通过激活JAK/STAT信号通路，诱导SV40MES13表达CXCL10 mRNA；TPL具有抑制IFN-γ诱导的SV40MES13表达CX-CL10的作用。%Objective To investigate the effects of triptolide (TPL) on the expression of CXCL10 mRNA induced by interferon gamma(IFN-γ) in mouse glomerular mesangial cells(SV40MES13) and its mechanism. Methods SV40MES13 were randomly divided into blank group,stimulation group,intervention group. Different concentrations of IFN-γ (0U/ml~2000U/ml) were used to stimulate the cells with different time(0h~48h),and then observe the changes of the expression of CXCL10 mRNA;CCK8 assay was used to de-tect the effects of different concentrations of TPL(2ng/ml~20ng/ml) on the survival rate of SV40MES13. The appropriate concentration of TPL causing cell survival rate>85%was applied to subsequent experiment,and then observe the effect of TPL on the expression of CXCL10 mRNA;JAK/STAT signaling pathway specific inhibitor AG490 was used to intervene in cells to investigate whether IFN-γinduced the expression of CXCL10 through the JAK/STAT signaling pathway. Real-Time PCR was used to detect the expression level of mRNA CXCL10. Results The present study showed that IFN-γinduced the expression of CXCL10 mRNA in SV40MES13 through a dose- and time-dependent manner;The JAK/STAT signaling pathway specific inhibitor AG490 inhibit the expression of CXCL10 mRNA induced by IFN-γin SV40MES13;TPL could inhibit the expression of CXCL10 mRNA induced by IFN-γ in SV40MES13 as well. Conclusion IFN-γ could induce expression of CXCL10 by activating JAK/STAT signaling pathway in SV40MES13;TPL could inhibit expression of CXCL10 by blocking JAK/STAT signaling pathway in SV40MES13.