HB-EGF-induced VEGF production and eNOS activation depend on both PI3 kinase and MAP kinase in HaCaT cells
HB-EGF-induced VEGF production and eNOS activation depend on both PI3 kinase and MAP kinase in HaCaT cells
复制标题
DOI：
10.1016/j.jdermsci.2009.06.002
复制
发表时间：
2009-09-01
影响因子：
4.6
通讯作者：
Kubota, Yasuo
中科院分区：
文献类型：
--
作者：
Nakai, Kozo;Yoneda, Kozo;Kubota, Yasuo
背景：肝素结合表皮生长因子样生长因子(HB-EGF)是一类与皮肤病理生理密切相关的生长因子。尽管内皮型一氧化氮合酶(ENOS)和血管内皮生长因子(VEGF)似乎参与了表皮角质形成细胞的有丝分裂和趋化作用，但HB-EGF诱导的eNOS和VEGF的激活及其信号转导机制尚不清楚。目的：探讨HB-EGF诱导人表皮角质形成细胞系(HaCaT细胞)eNOS激活和VEGF产生的可能信号转导途径。采用酶联免疫吸附试验检测血管内皮细胞生长因子的产生。结果：HB-EGF可诱导EGFR的磷酸化，并在1h达到峰值，HB-EGF可在几分钟内诱导p42/p44man的磷酸化。P42/p44MAPK抑制剂U0126和磷脂酰肌醇3-激酶(PI3K)抑制剂LY294002可显著阻断HB-EGF诱导的eNOS活化。HB-EGF促进血管内皮生长因子的产生。U0126和LY294002可阻断HB-EGF诱导的血管内皮生长因子的产生。结论：HB-EGF诱导HaCaT细胞eNOS活化依赖于p42/p44man、PI3K/Akt通路和内源性VEGF。(C)2009年日本皮肤病研究学会。爱思唯尔爱尔兰有限公司出版。保留所有比赛。
Background: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of growth factors that have been implicated in skin patho-physiology. Although endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) appear to be involved in mitogenesis and chemotaxis in epidermal keratinocytes, the activation of eNOS and VEGF production induced by HB-EGF and its signaling mechanism remains undefined.Objective: We examined possible signal transduction pathways by which HB-EGF leads to eNOS activation and VEGF production in human epidermal keratinocyte cell line (HaCaT cells).Methods: The phosphorylation of epidermal growth factor receptor (EGFR), mitogen-activated Protein kinase (MAPK; p42/p44 MAPK), Akt and eNOS were examined by Western blotting analysis. VEGF production was determined by enzyme-linked immunosorbent assay. Various inhibitors were utilized to investigate the signaling mechanisms of eNOS activation and VEGF production.Results: HB-EGF-induced phosphorylation of EGFR with maximum phosphorylation at I h. HB-EGF-induced phosphorylation of p42/p44 MAN in a few minutes. It activated Akt with maximum phosphorylation at I h and eNOS with maximum phosphorylation at 3 h. The HB-EGF-induced eNOS activation was significantly blocked by the p42/p44 MAPK inhibitor U0126 and the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002. HB-EGF increased VEGF production. The HB-EGF-induced VEGF production was blocked by U0126 and LY294002. Finally, the HB-EGF-induced activation of Akt and eNOS was Suppressed by VEGF competitive antagonist, CBO-P11.Conclusion: These results demonstrate that HB-EGF-induced eNOS activation depends on p42/p44 MAN, PI3K/Akt pathways and endogenous VEGF in HaCaT cells. (C) 2009 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All Fights reserved.