摘要
目的观察苦参素对脂多糖（LPS）诱导下人肾小球系膜细胞（HMC）增殖时磷酸化信号传导和转录激活因子3（p-STAT3）、活化的信号传导和转录激活因子3蛋白抑制剂（PIAS3）蛋白和mRNA表达的影响，并探讨其相互关系。方法体外培养HMC，并分为对照组、LPS模型组及苦参素干预组。培养12、24、48h时以MTF法检测HMC的增殖情况：ELISA法检测细胞上清Ⅳ型胶原蛋白（CollV）含量；同时收集同时间点细胞，采用Western印迹法检测p-STAT3和PIAS3的蛋白表达；实时荧光定量PCR法检测STAT3和PIAS3的mRNA表达。结果LPS组细胞增殖较对照组显著加快（P〈0．01），CollV的表达显著高于对照组（P〈0．01）。苦参素干预后细胞增殖和ColIV的表达显著低于模型组（P〈0．01）。LPS诱导12h时细胞p-STAT3表达开始上调，各时间点P—STAT3表达量显著高于对照组（P〈0，01）。苦参素干预后，与同时间点LPS模型组相比显著下调（P〈0．01）。LPS诱导下PIAS3各时间点表达量显著下调（P〈0．01）。苦参素干预后，与同时间点LPS模型组相比均显著上调（P〈0．01）。结论苦参素对LPS诱导HMC增殖过程中p-STAT3蛋白及mRNA表达有下调作用，对PIAS3有上调作用。苦参素可能影响HMC增殖过程中JAK—STAT信号传导通路。
Objective To study the effect of oxymatrine on p-STAT3 and PIAS3 signaling molecule and it＇s mRNA expression in the proliferation of the human mesangial cells （HMCs） induced by lipopolysaccharide（LPS） and to explore their relationship. Methods HMCs were divided into three groups： control group, LPS group and oxymatrine group. HMC proliferation was detected by methyl thiazolyl tetrazolium （MTF） assay. Type Ⅳcollagen in the supernatants of the cultured HMCs was detected by ELISA at 12, 24, 48 hours respectively. At the same time, the protein and mRNA expressions of p-STAT3 and PIAS3 were measured by Western blot and realtime quantitative RT-PCR. Results The cell proliferation, the mRNA and protein expression of type Ⅳ collagen, p-STAT3 in LPS group were increased compared with the control group （P〈0.01）, but they were decreased in oxymatrine group （P 〈 0.01）. The expressions of protein and mRNA of PIAS3 in LPS group were decreased significantly compared with control group （P〈0.01）, but they were increased in oxymatrine group （P〈0.01）. Conclusion Oxymatrine can down-regulate the expression of p-STAT3 and up-regulate the expression of PIAS3, which plays an important role in the process of LPS-induced HMCs proliferation.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2009年第8期635-639,共5页
Chinese Journal of Nephrology
基金
甘肃省中青年自然科学基金（YS-011-A23-022）
甘肃省中医药科研项目（GZK-2008-5）
