Interferon-γ enhances superoxide production in human mesangial cells via the JAK-STAT pathway
Interferon-γ enhances superoxide production in human mesangial cells via the JAK-STAT pathway
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DOI：
10.1038/sj.ki.5001639
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发表时间：
2006-08-01
影响因子：
19.6
通讯作者：
Kohno, M.
中科院分区：
文献类型：
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作者：
Moriwaki, K.;Kiyomoto, H.;Kohno, M.
免疫反应性细胞因子，如干扰素（IFN）-γ，在肾小球肾炎中具有多种作用。与肾小球肾炎进展相关的超氧阴离子（O-2（-））主要由烟酰胺腺嘌呤二核苷酸磷酸（还原型）NAD（P）H氧化酶产生。本实验观察了IFN-γ对人肾小球系膜细胞（HMCs）O-2（-）产生、信号转导和转录激活因子（STAT）-1 α磷酸化、NAD（P）H氧化酶组成部分p22 phox和Nox 1 mRNA和蛋白表达的影响。含黄素的酶抑制剂diphenyleneiodonium（10 μ mol/l）和Janus激活激酶（JAK）2抑制剂AG 490（100 μ mol/l）完全消除了IFN-γ引起的O-2（-）产生的显著增加。IFN-γ刺激5 min后，用Western印迹分析检测到磷酸化STAT-1 α，并从30 min至4 h观察到与γ激活位点的结合，此后通过电泳迁移率变动分析（EMSA）观察。EMSA的超移位分析显示主要转录因子为STAT-1 α。IFN-γ显著增加p22 phox mRNA和蛋白的表达，尽管表达被AG 490抑制。这些数据表明IFN-γ通过JAK-STAT途径和NAD（P）H氧化酶刺激HMCs中O-2（-）的产生。
Immune reactive cytokines, such as interferon (IFN)-gamma, have multiple effects in glomerulonephritis. Superoxide anions (O-2(-)), which are associated with the progression of glomerulonephritis, are mainly generated by nicotinamide adenine dinucleotide phosphate ( reduced form) NAD(P)H oxidases. We determined the effects of IFN-gamma on O-2(-) production, phosphorylation of signal transducer and activator of transcription (STAT)-1 alpha, and the mRNA and protein expressions of p22phox and Nox1, components of NAD( P) H oxidases, in human mesangial cells (HMCs). Significant increases in O-2(-) production with IFN-gamma were completely abolished by the flavin-containing enzyme inhibitor, diphenyleneiodonium ( 10 mu mol/l), and the Janus-activated kinase (JAK)2 inhibitor, AG490 (100 mu mol/l). Phosphorylated STAT-1 alpha was detected after 5min of IFN-gamma stimulation using Western blot analysis, and binding to the gamma-activating site was observed from 30min to 4 h, thereafter by electrophoretic mobility shift assay ( EMSA). Super-shift analysis in EMSA revealed that the main transcription factor was STAT-1 alpha. IFN-gamma significantly increased the expression of p22phox mRNA and protein, although expression was inhibited by AG490. These data suggest that IFN-gamma stimulates O-2(-) production in HMCs via the JAK-STAT pathway and NAD(P) H oxidase.