苦参素对LPS诱导的人肾小球系膜细胞中p-STAT3和SOCS-3的影响

Effect of Oxymatrine on expression of p-STAT3 and SOCS3 in lipopolysaccharide-induced human glomerularmesangial cells

目的:观察苦参素对LPS诱导的人肾小球系膜细胞(HMC)中磷酸化信号转导和转录活化因子3(p-STAT3)和细胞因子信号抑制因子3(SOCS3)的表达及相互关系.方法:体外原代培养的HMC,分为正常组、LPS(10 mg/L)组、LPS(10mg/L)+苦参素(320 mg/L)组,每组分3个时间点(12,24,48h).采用MTT法检测HMC的增殖,RT-PCR方法检测STAT3和SOCS3 mRNA的表达,Western Blot方法检测p-STAT3和SOCS3蛋白的表达.结果:苦参素能明显抑制LPS诱导的HMC增殖,与正常组相比,p-STAT3和STAT3 mRNA的表达在12,24,48 h均上调,SOCS3蛋白和mRNA的表达也同步上调.与LPS诱导组相比,苦参素组p-STAT3和STAT3 mRNA的表达在12,24,48 h均下调,而SOCS3蛋白和mRNA表达有明显上调的趋势.结论:苦参素能抑制LPS诱导的HMC的增殖,且能上调LPS诱导的HMC中SOCS3 mRNA和蛋白表达,下调P-STAT3和STAT3 mRNA的表达,其作用可能与上调SOCS3、抑制STAT3磷酸化有关.

AIM： To observe the effects of Oxymatfine on the phosphorylation expression of signal transducers and activators of transcription 3 （ p-STAT3 ） and suppressors of cytokine signaling 3 （ SOCS3 ） in lipopolysaceharide （LPS） -induced human glomerularmesangial cells （ HMGs ）. METHODS ： HMCs cultured in vitro were divided into 3 groups： control group, LPS （10 mg/L） group and OM group （ LPS 10 mg/L ＋ Oxymatfine 320 mg/L）. HMCs proliferation was detected by MTT assay, protein expressions of p-STAT3 and SOCS3 were observed by Western blot, and STAT3 and SOCS3 mRNA were measured by real-time quantitative RT-PCR. RESULTS： Compared with those in normal control group, the expressions of p-STAT3, STAT3 mRNA, SOCS3 and SOCS3 mRNA all significantly increased in PS-induced HMCs. Compared with those in LPS group, the expressions of p-STAT3 and STAT3 mRNA decreased in OM group, but the expression changes of SOCS3 and SOCS3 mRNA displayed an increasing tendency. CONCLUSION： Oxymarine inhibits the proliferation of PS-indueed HMCs and increases the expressions of SOCS3 and SOCS3 mRNA. Oxymarine also decreases the expressions of p-STAT3 and STAT3 mRNA, possibly by increasing SOCS3 to inhibit the expressions of p-STAT3.