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目的 探讨脐带静脉内皮细胞在缺氧条件培养下血管内皮生长因子特异受体的表达 ,为进一步研究血管内皮生长因子及其受体在新血管生成中的作用提供理论依据。方法 体外培养脐带静脉内皮细胞 ,不同缺氧时间处理及血管活性因子刺激 ,提取组织总RNA ,应用Northern杂交和逆转录聚合酶连扩增反应 (RT PCR)等方法 ,观察基因表达的变化。结果 内皮细胞无缺氧和缺氧 3h均未见flt 1表达 ,缺氧 6h可见flt 1摸板核糖核酸 (mRNA)的表达。内皮细胞缺氧 6h ,内皮素 (ET)、一氧化氮 (NO)、LNNA对flt 1表达有影响。ET促进flt 1mRNA的表达 ,NO抑制其表达 ,而LNNA部分恢复其表达。在无缺氧和缺氧 3,6hKDR基因均有表达。内皮细胞缺氧 6h ,ET、NO、LNNA对KDR无影响。LNNA阻断了内源性NO后 ,明显促进KDRmRNA的表达 ,其他无作用。结论 在缺氧条件下血管内皮细胞生长因子的受体flt 1表达明显增加并受血管活性因子调节 ,KDR有表达且部分受血管活性因子调节
Objective To investigate the expression of vascular endothelial growth factor (VEGF) -receptor in umbilical vein endothelial cells cultured under hypoxic conditions, and provide a theoretical basis for further study on the role of VEGF and its receptor in neovascularization. Methods The umbilical vein endothelial cells were cultured in vitro, treated with different time of hypoxia and stimulated by vasoactive factors. The total RNA was extracted and the changes of gene expression were observed by Northern blotting and reverse transcriptase polymerase chain reaction (RT PCR). Results No expression of flt 1 was found in endothelial cells exposed to hypoxia and hypoxia for 3 h. Flt 1 mRNA expression was detected in hypoxia for 6 h. Endothelial cells hypoxia 6h, endothelin (ET), nitric oxide (NO), LNNA flt 1 expression. ET promotes the expression of flt1mRNA, NO inhibits its expression, whereas LNNA partially regains its expression. No hypoxia and hypoxia 3,6 hKDR gene expression. Endothelial cells hypoxia 6h, ET, NO, LNNA had no effect on KDR. After LNNA blocked the endogenous NO, the expression of KDR mRNA was obviously promoted, while the others did not. Conclusions Under hypoxic conditions, the expression of vascular endothelial growth factor receptor flt 1 is significantly increased and regulated by vasoactive factors, KDR is expressed and partially regulated by vasoactive factors