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作 者:邹丽萍[1] 蒋涛[1] 车祺[1] 李慧[1] 瞿俊杰[1] 陈琦[1] 张农[1]
机构地区:[1]复旦大学上海医学院病理学系,上海200032
出 处:《复旦学报(医学版)》2007年第4期557-562,共6页Fudan University Journal of Medical Sciences
基 金:国家自然科学基金项目(30170430)
摘 要:目的探讨醛糖还原酶抑制剂(ARI)影响血小板源性生长因子-BB(PDGF-BB)促体外培养大鼠系膜细胞(MsC)增殖作用的机制。方法用四甲基偶氮唑盐(MTT)比色法检测细胞增殖,流式细胞术分析细胞周期,对比PDGF-BB刺激MsC生长速度与MAPK3条信号通路(ERK、JNK及p38)抑制剂及PI3K信号通路抑制剂对PDGF促MsC增殖作用的影响。用半定量RT-PCR检测醛糖还原酶抑制剂(ARI)对MsC合成内源性PDGF-B链的影响。蛋白印迹法观察ARI对PDGF引起的信号通路磷酸化的影响。结果(1)ERK抑制剂U0126、JNK抑制剂SP600125及PI3K抑制剂LY294002能明显抑制PDGF对MsC的促增殖作用(P<0.05),而p38抑制剂SB203580不能抑制PDGF的促MsC增殖作用;(2)ARI在转录水平对MsC合成内源性PDGF-B链没有明显影响。(3)PDGF刺激MsC后引起ERK、JNK及PI3K/Akt信号通路的磷酸化,而ARI仅能抑制JNK及PI3K/Akt两条信号通路的磷酸化(P<0.05),对ERK信号通路的磷酸化没有明显影响。结论ARI部分抑制PDGF促MsC增殖作用可能与其影响JNK和PI3K/Akt信号通路活化有关。Purpose To study the mechanism of effects of aldose reductase inhibitors on rat mesangial cells (MsC) in vitro proliferation induced by PDGF-BB. Methods Cell proliferation was assessed by MTT colorimetric assay. Cell cycle and apoptosis were analyzed by flow cytometry. The growth of MsC proliferation stimulated by PDGF,with or without respective inhibitor of ERK,JNK,p38 and PI3K,was compared. The impact of aldose reductase inhibitor (ARI) on PDGF-B chain transcription level of MsC was assessed by semi-quantitative RT-PCR. The influence of ARIs on the activation of MAPK and PI3K/Akt signal transduction pathways induced by PDGF was measured by Western blot. Results The inhibitors of ERK,JNK and PI3K,but not p38,greatly reduced the MsC proliferation (P 〈0.05) stimulated by PDGF. ARIs did not apparently change the transcription (P〉0.05) of PDGF-B chain of MsC. ARIs could prevent the PDGF-induced phosphorylation of JNK and PI3K/Akt, but not ERK. Conclusions ARIs may partially reduce the PDGF-induced proliferation of MsC through JNK and PI3K/Akt signal transduction pathway.
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