Anti‐apoptotic action of hepatocyte growth factor through mitogen‐activated protein kinase on human aortic endothelial cells
Anti‐apoptotic action of hepatocyte growth factor through mitogen‐activated protein kinase on human aortic endothelial cells
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肝细胞生长因子通过丝裂原激活蛋白激酶对人主动脉内皮细胞的抗凋亡作用
DOI：
10.1097/00004872-200018100-00008
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发表时间：
2000
影响因子：
4.9
通讯作者：
T. Ogihara
中科院分区：
文献类型：
--
作者：
H. Nakagami;R. Morishita;Kei Yamamoto;Y. Taniyama;M. Aoki;Shokei Kim;Kunio Matsumoto;Toshikazu Nakamura;J. Higaki;T. Ogihara
目的探讨肝细胞生长因子(HGF)抗血管内皮细胞凋亡的分子机制。方法采用水溶性四唑盐染色法、DNA片段酶联免疫吸附试验、Hoechst 33258和碘化丙啶非共聚焦荧光显微镜核染色法测定细胞数量和死亡指数。细胞外信号调节蛋白激酶(ERK)和p38丝裂原活化蛋白激酶(P38MAPK)用磷酸化抗体进行Western blotting分析。结果HGF处理静止期内皮细胞后，细胞数呈剂量依赖性增加，乳酸脱氢酶(LDH)释放量减少。此外，HGF显著减轻无血清条件下培养所致的内皮细胞死亡。因此，我们重点研究了信号转导系统，特别是ERK和p38MAPK。ERK被HGF明显磷酸化。加入MAPK的特异性抑制剂PD98059，以剂量依赖的方式显著减弱HGF诱导的内皮细胞数量的增加，这支持了ERK对细胞生长的贡献。同样，在无血清条件下，PD98059也可减轻HGF对LDH释放和DNA片段化的影响。有趣的是，ERK在刺激后12小时重新磷酸化。ERK的重新磷酸化是外源性HGF诱导内源性HGF的结果，在条件培养液中加入中和的抗HGF抗体可减弱12h时ERK的再次磷酸化，相反，尽管p38MAPK也被HGF磷酸化，但p38MAPK的特异性抑制剂SB203580不能改变HGF诱导的内皮细胞生长。结论HGF抗内皮细胞死亡的作用主要是通过对人内皮细胞ERK的磷酸化实现的。
Objective To investigate the molecular mechanisms of the anti-apoptotic action of hepatocyte growth factor (HGF), a novel angiogenic growth factor that may have a pivotal role in the regulation of endothelial cells, on human aortic endothelial cells. Methods An index of cell number and death was determined using a water-soluble tetrazolium salt dye assay, DNA fragmentation enzyme-linked immunosorbent assay, and non-confocal fluorescence microscopy of nuclear staining with Hoechst 33258 and propidium iodide. Extracellular-signal-regulated protein kinase (ERK) and the p38 mitogen-activated protein kinase (p38MAPK) were analysed by Western blotting using a phospho-specific antibody. Results Treatment of quiescent endothelial cells with HGF resulted in significant dose-dependent increases in cell numbers and decreases in lactate dehydrogenase (LDH) release. Moreover, HGF significantly attenuated endothelial cell death induced by culture in serum-free conditions. We therefore focused on the signal transduction system, and in particular on ERK and p38MAPK. ERK was markedly phosphorylated by HGF. The contribution of ERK to cell growth was supported by the observation that addition of PD98059, a specific inhibitor of MAPK kinase, significantly attenuated the increase in endothelial cell numbers induced by HGF, in a dose-dependent manner. Similarly, PD98059 also attenuated the decrease in LDH release and DNA fragmentation by HGF under serum-free conditions. Interestingly, ERK was re-phosphorylated at 12 h after stimulation. Re-phosphorylation of ERK was the result of induction of endogenous HGF by exogenously added HGF, as addition of neutralizing anti-HGF antibody to the conditioned medium attenuated re-phosphorylation of ERK at 12 h. In contrast, although p38MAPK was also phosphorylated by HGF, SB203580, a specific inhibitor of p38MAPK, failed to change the endothelial cell growth induced by HGF. Conclusion We have demonstrated that the anti-apoptotic action of HGF against endothelial cell death was mainly through phosphorylation of ERK on human endothelial cells.
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肝细胞生长因子受体 c-met 的信号转导。
DOI：
10.1681/asn.v5111872
发表时间：
1995
期刊：
Journal of the American Society of Nephrology : JASN
影响因子：
--
作者：
Cantley,LG;Cantley,LC
通讯作者：
Cantley,LC
DOI：
10.1056/nejm199405193302008
发表时间：
1994-05
期刊：
The New England journal of medicine
影响因子：
--
作者：
G. Gibbons;V. Dzau
通讯作者：
G. Gibbons;V. Dzau
机械和体液因素在血管平滑肌和心肌细胞生长调节中的作用。
影响因子：
3.2
作者：
Dzau,VJ
通讯作者：
Dzau,VJ
影响因子：
3.7
作者：
Gupta, K;Kshirsagar, S;Hebbel, RP
通讯作者：
Hebbel, RP
影响因子：
4.8
作者：
Chen, YR;Wang, XP;Tan, TH
通讯作者：
Tan, TH