氧化苦参碱对脂多糖诱导的人肾小球系膜细胞增殖p-STAT1/PIAS1信号分子的影响  

Oxymatrine suppresses p-STAT1/PIAS1 signaling in LPS-induced human mesangial cells proliferation

作　　者：党红星[1] 金玉[1] 李宇宁[1] 凌继祖[1] 苏婕[1] 

机构地区：[1]兰州大学第一医院儿科,兰州730000

出　　处：《第三军医大学学报》2009年第17期1645-1648,共4页Journal of Third Military Medical University

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)~~

摘　　要：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 effect of oxymatrine （OM） on the expressions of p-STAT1 and PIAS1 signaling molecules at protein and mRNA levels in the proliferation of the human mesangial cells （HMC） induced by lipopolysaccharide （LPS） and explore the relationship between them. Methods HMCs were primarily cultured from a 4-month-old aborted human fetus （with informed consent and approved by the Ethics Committee of Lanzhou University）, and then divided into 3 groups, that is, control group , LPS group （10 ng/ml） and OM group （ LPS 10 ng/ml and OM 320 mg/L）. After cultured for 12, 24 and 48 h respectively, HMC proliferation were analyzed by methyl thiazolyl tetrazolium （MTT） assay and type Ⅳ collagen in the supernatants were detected by ELISA. At the same time points, the cells lysates were collected for the mRNA and protein expressions of p-STAT1 and PIAS1 by real-time quantitative RT-PCR and Western blot analysis. Results The cell proliferation of LPS group was faster and the type Ⅳ collagen protein was increased more than the control group （P 〈0.01 ） ,but decreased in OM treatment group （P 〈0.01 ） ; The protein expression of p-STAT1 and its mRNA in LPS group were upregulated compared with the control group （P 〈 0.01 ）, but decreased in OM treatment group at the same time points （P 〈 0. 01 ） ; the expression of protein and mRNA of PIAS1 were lower than control group significantly （ P 〈 0.01 ）, but become higher level in OM treatment group （ P 〈 0.01 ）. Conclusion OM downregulated the increased expressions of p-STAT1/PIAS1 in the process of LPS-induced HMC proliferation, probably through the JAK/STAT signal pathway.

关 键 词：人肾小球系膜细胞 增殖 氧化苦参碱 JAK/STAT信号通路 p—STAT1 PIAS1分子 

分 类 号：R282.5[医药卫生—中药学] R322.61[医药卫生—中医学]