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用噻唑蓝比色法 (MTT法 )、H3 胸腺嘧啶核苷 (H3 TdR)掺入法和流式细胞术 ,观察红细胞生成素 (EPO)3’端增强子片段对培养的猪肺动脉平滑肌细胞 (PASMCs)的内皮依赖性和非内皮依赖性低氧性增殖的影响。结果为 :(1)低氧 2 4h后PASMCs明显增殖 ,转入野生型EPO3’端增强子片段可被抑制 ,而转入突变型片段无此作用 ;(2 )肺动脉内皮细胞 (PAECs)低氧 2 4h ,其条件培养液有明显的促PASMCs增殖作用 ,将野生型EPO3’端增强子片段先转入PAECs,此作用明显减弱 ,而转入突变型片段则无明显影响。提示 :(1)PAECs低氧条件培养液可促进PASMCs增殖 ,PASMCs也可直接感受低氧而增殖 ,PAECs和PASMCs的低氧反应均被外源性EPO3’增强子片段抑制 ;(2 )由于EPO3’增强子上有低氧诱导因子 1(HIF 1)结合位点 ,PAEC和PASMC低氧反应可能是通过HIF 1低氧信号转导的共同通路
The effects of the 3 ’enhancer fragment of erythropoietin (EPO) on the proliferation of cultured porcine pulmonary artery smooth muscle cells (HUVECs) were observed by MTT method, H3TdR incorporation method and flow cytometry PASMCs) in endothelial-dependent and non-endothelial-dependent hypoxic proliferation. The results were as follows: (1) The PASMCs proliferated significantly after hypoxia for 24 hours, and the transfected wild-type EPO 3 ’end enhancer fragment could be inhibited, but the mutant fragment did not. (2) The pulmonary arterial endothelial cells (PAECs) At 24 hours, the conditioned medium had a significant effect on the proliferation of PASMCs. The wild-type EPO 3 ’enhancer fragment was first transferred to PAECs and the effect was obviously weakened. However, no mutation was found in the medium. The results suggest that: (1) PAECs hypoxic conditions can promote the proliferation of PASMCs, PASMCs can also directly feel hypoxia and proliferation, PAECs and PASMCs hypoxia response were inhibited by exogenous EPO3 ’enhancer fragment; (2) As EPO3 ’Hypoxia-inducible factor 1 (HIF 1) binding site on the enhancer, the hypoxia response of PAEC and PASMC may be through the common pathway of HIF 1 hypoxia signaling