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目的观察氧化苦参碱对LPS诱导下HMC增殖时p-STAT1、PIAS1蛋白和mRNA表达情况,探讨其相互关系。方法体外培养HMC,分为空白对照组和LPS模型组及氧化苦参碱干预组,培养12、24、48 h时以MTT法检测HMC的增殖情况,ELISA检测细胞上清Col-Ⅳ含量,收集同时段细胞,提取细胞裂解蛋白和mRNA,采用Western blot检测p-STAT1和PIAS1的蛋白表达,实时荧光定量PCR检测STAT1和PIAS1 mRNA表达。结果LPS组细胞增殖较对照组显著加快(P<0.01),Col-Ⅳ的表达显著高于对照组(P<0.01);氧化苦参碱干预后细胞增殖和Col-Ⅳ的表达显著较模型组降低(P<0.01)。蛋白和mRNA表达情况:LPS诱导下各时间段p-STAT1表达升高,表达量显著高于对照组(P<0.01),经OM干预后表达显著下降(P<0.01);LPS诱导下PIAS1表达量显著降低(P<0.01),经OM干预,与LPS组相比各时间段均显著上升(P<0.01)。结论氧化苦参碱对LPS诱导的人系膜细胞增殖过程中p-STAT1/PIAS1蛋白及mRNA表达有调节作用,氧化苦参碱可能影响JAK/STAT信号转导通路。
Objective To observe the expression of p-STAT1 and PIAS1 protein and mRNA in oxymatrine (HMC) -induced LPS-induced HMC proliferation and to explore their relationship. Methods HMC were cultured in vitro and divided into blank control group, LPS model group and oxymatrine intervention group. The proliferation of HMC was detected by MTT assay at 12, 24 and 48 h after culture. Col-Ⅳ content in supernatant was detected by ELISA. At the same time, cells were harvested for cell lysis of protein and mRNA. The protein expression of p-STAT1 and PIAS1 was detected by Western blot. The expression of STAT1 and PIAS1 mRNA was detected by real-time fluorescence quantitative PCR. Results The proliferation of LPS group was significantly higher than that of the control group (P <0.01), and the expression of Col-Ⅳ was significantly higher than that of the control group (P <0.01). Compared with the model group Decreased (P <0.01). (P <0.01). The expression of PIAS1 in LPS group was significantly lower than that in control group (P <0.01), and the expression of PIAS1 was down-regulated by LPS (P <0.01) (P <0.01). Compared with LPS group, the levels of OM increased significantly (P <0.01). Conclusion Oxymatrine can regulate the expression of p-STAT1 / PIAS1 protein and mRNA during LPS-induced human mesangial cell proliferation. Oxymatrine may affect JAK / STAT signal transduction pathway.