Hypoxic stimulation of vascular endothelial growth factor expression in activated rat hepatic stellate cells
Hypoxic stimulation of vascular endothelial growth factor expression in activated rat hepatic stellate cells
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DOI：
10.1002/hep.510310122
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发表时间：
2000-01-01
期刊：
影响因子：
13.5
通讯作者：
Dai, ZH
中科院分区：
文献类型：
--
作者：
Ankoma-Sey, V;Wang, Y;Dai, ZH
伤口愈合期间对缺氧刺激的组织修复反应包括血管生成因子（例如血管内皮生长因子（VEGF））的产生增加。肝星状细胞是氧敏感细胞，能够产生VEGF。我们假设，缺氧刺激信号在活化的星状细胞介导VEGF分泌在肝损伤。具体目的是评估缺氧对HSC-T6细胞（一种永生化大鼠肝星状细胞系）中VEGF基因表达的影响，并在大鼠原代培养的星状细胞中，VEGF mRNA的缺氧诱导具有剂量和时间依赖性。低氧刺激VEGF信使RNA（mRNA）与低氧T6细胞条件培养基中VEGF蛋白的分泌相关。S-亚硝基-N-乙酰基-D，L-青霉胺（SNAP），一氧化氮（NO）供体，去铁胺（DFx）和氯化钴，模拟细胞缺氧，类似地刺激VEGF mRNA的表达和分泌。在常氧或低氧激活的星状细胞中检测到四种先前描述的VEGF mRNA的剪接变体（VEGF-120、144、164、188）。在缺氧的T6细胞中，VEGF受体Fit-1和Flk-1的表达存在差异。低氧条件选择性刺激Fit-1 mRNA表达，而Flk-1 mRNA保持不变。在原代星状细胞培养物和体内损伤后，也证实了VEGF的低氧诱导，低氧刺激星状细胞中的细胞信号传导，最终快速诱导VEGF和Flt-1 mRNA表达和VEGF分泌。VEGF的低氧诱导被NO模拟，并且可能在肝创伤愈合和肝癌发生的发病机制中具有重要的机制。
The tissue repair response to hypoxic stimuli during wound healing includes enhanced production of angiogenic factors, such as vascular endothelial growth factor (VEGF). Hepatic stellate cells are oxygen-sensing cells, capable of producing VEGF. We hypothesized that hypoxia-stimulated signaling in activated stellate cells mediate VEGF secretion during liver injury. The specific aim was to evaluate the effect of hypoxia on the gene expression of VEGF in HSC-T6 cells, an immortalized rat hepatic stellate cell line, and in rat primary cultures of stellate cells, Hypoxic induction of VEGF mRNA was dose- and time-dependent. The hypoxic stimulation of VEGF messenger RNA (mRNA) correlated with the secretion of VEGF protein in conditioned media by hypoxic T6 cells. S-Nitroso-N-acetyl-D,L-penicillamine (SNAP), a nitric oxide (NO) donor, and desferrioxamine (DFx) and cobalt chloride, mimics of cellular hypoxia, similarly stimulated VEGF mRNA expression and secretion. Four previously described splice variants of the VEGF mRNA (VEGF-120, 144, 164, 188) were detected in both normoxic- or hypoxic-activated stellate cells. There was differential expression of the VEGF receptors, Fit-1 and Flk-1, in hypoxic T6 cells. Hypoxic conditions selectively stimulated Fit-1 mRNA expression, whereas Flk-1 mRNA remained unchanged. Hypoxic induction of VEGF was also demonstrated in primary stellate cell cultures and after in vivo injury, Hypoxia stimulates cell signaling in stellate cells, culminating in the rapid induction of VEGF and Flt-1 mRNA expression and VEGF secretion, The hypoxic induction of VEGF is mimicked by NO and may be of mechanistic importance in the pathogenesis of hepatic wound healing and hepatocarcinogenesis.