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体外合成红细胞生成素(Epo)3’-端增强子野生型及突变型片段,借助脂质体转入培养的大鼠肺微血管内皮细胞,用半定量RT-PCR方法测定内皮生长因子(VEGF)mRNA。结果发现:①内皮细胞在常氧下培养有VEGF基因转录,mRNA量吸光度(A)值为2.32±0.36;②缺氧4h时VEGF基因转录增加,mRNA量(A)值为4.71±0.52,与常氧组比较PO.05。结果提示:在VEGF基因序列中可能存在Epo 3’-端增强子片段,它参与了内皮细胞的缺氧反应。
The wild type and mutant fragments of erythropoietin (Epo) 3’-end enhancer were synthesized in vitro and transfected into cultured rat pulmonary microvascular endothelial cells by liposomes. The expression of VEGF was measured by semi-quantitative RT-PCR. mRNA. The results showed that: (1) endothelial cells cultured under normoxia have VEGF gene transcription, the mRNA absorbance (A) value was 2.32 ± 0.36; (2) VEGF mRNA transcription increased after 4h hypoxia, mRNA value (A) was 4.71 ± 0.52, Compared with normoxia group, P <0.05; (3) Wild-type Epo 3’-terminal enhancer fragment can block the induction of VEGF gene transcription by hypoxia, the mRNA level (A) was 2.29 ± 0.41, P <0.05; while the mutant fragment had no effect, the amount of VEGF mRNA (A) was 4.57 ± 0.57, compared with the hypoxia group (P> 0.05). The results suggest that there may be an Epo 3’-end enhancer fragment in the VEGF gene sequence, which is involved in endothelial cell hypoxia.