Regulation of VEGF mRNA expression and protein secretion by TGF-β2 in human retinal pigment epithelial cells
Regulation of VEGF mRNA expression and protein secretion by TGF-β2 in human retinal pigment epithelial cells
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DOI：
10.1016/j.exer.2006.12.016
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发表时间：
2007-05-01
影响因子：
3.4
通讯作者：
Elner, Victor M.
中科院分区：
文献类型：
--
作者：
Bian, Zong-Mei;Elner, Susan G.;Elner, Victor M.
人类视网膜色素上皮（HRPE）的VEGF分泌在视网膜和脉络膜新生血管形成中起重要作用。在这项研究中，在HRPE细胞中研究了转化生长因子-BETA 2（TGF-β2）诱导的血管内皮生长因子（VEGF）基因表达。用TGF-β2治疗HRPE细胞为24和48 h，而8小时则显着增加了VEGF的分泌，分别增加了五倍和九倍。诱导的VEGF mRNA在刺激后的3小时内达到峰值，并在36小时保持基础上方。 TGF-β2刺激VEGF表达被环己酰亚胺阻断，这表明需要从头蛋白质合成。抗燃料燃料，地塞米松和环孢菌素A的诱导VEGF产生强烈抑制。尽管单独使用TNF-Alpha或BFGF刺激VEGF表达的刺激较弱，但这两种细胞因子中的任何一种都使TGF-BETA 2的效应促进了TGF-Beta 2的作用。关于VEGF mRNA表达和蛋白质的产生。定量RT-PCR表明该协同作用主要是在VEGF转录水平上。此外，TGF-beta 2诱导的RPE VEGF分泌显着降低了有丝分裂原激活蛋白（MAP）激酶（MEK）（MEK）（UO126）（UO126），p38（SB202190），C-Jun NH2-NH2-末端激酶（JNK），Sp600125，c-Jun NH2-末端激酶（SB202190），蛋白酪氨酸激酶（PTK）（染料木黄酮）和pbosphatidylylinininolininolininyl 3-激酶（PI3K）（LY294002）。蛋白激酶C（PKC）（RO318220），核因子-Kappa B（NF-KAPPA B）[咖啡酸苯乙酯酯（CAPE）]和活性氧（ROS）[N--乙酰基形成碱（NAC）和二苯基二烯（DPI）]。这些结果表明MEK，p38，JNK，PI3K和NF-KAPPA B以及包括PKC，PTK和ROS在内的多个必需信号中间体，由TGF-Beta 2。有限公司保留所有权利。
VEGF secretion by the human retinal pigment epithelium (hRPE) plays an important role in retinal and choroidal neovascularization. In this study, transforming growth factor-beta 2 (TGF-beta 2)-induced vascular endothelial growth factor (VEGF) gene expression was investigated in hRPE cells. Treatment of hRPE cells with TGF-beta 2 for 24 and 48 h as compared to 8 h resulted in markedly increased VEGF secretion by fivefold and nine-fold, respectively. Induced VEGF mRNA peaked within 3 h of stimulation and remained above the basal at 36 h. Stimulation of VEGF expression by TGF-beta 2 was blocked by cycloheximide, suggesting that de novo protein synthesis is required. Induced VEGF production was strongly inhibited by anti-in flammatoryagents, dexamethasone and cyclosporin A. Despite of the weak stimulation of VEGF expression by TNF-alpha or bFGF alone, co-administration of either of these two cytokines synergized the effect of TGF-beta 2 on VEGF mRNA expression and protein production. Quantitative RT-PCR revealed that the synergy was predominantly at the level of VEGF transcription. Moreover, TGF-beta 2-induced RPE VEGF secretion was significantly reduced by inhibitors of mitogen-activated protein (MAP) kinase (MEK) (UO126), p38 (SB202190), c-Jun NH2-terminal kinase (JNK), Sp600125, protein tyrosine kinase (PTK) (Genistein), and pbosphatidylinositol 3-kinase (PI3K) (Ly294002). Induced VEGF expression was completely abrogated by inhibitors of protein kinase C (PKC) (Ro318220), nuclear factor-kappa B (NF-kappa B) [caffeic acid phenethyl ester (CAPE)], and reactive oxygen species (ROS) [N-acetyl-cysteine (Nac) and diphenyleneiodonium (DPI)]. These results suggest that MEK, p38, JNK, PI3K, and NF-kappa B as well as multiple essential signaling intermediates, including PKC, PTK and ROS, are involved in hRPE VEGF up regulation by TGF-beta 2. (C) 2006 Elsevier Ltd. All rights reserved.