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目的 探讨缺氧对大鼠肺动脉平滑肌细胞 (PASMCs)信号转导及转录活化因子 3(STAT 3)表达及细胞增殖的影响。方法 采用组织块法原代培养PASMCs并进行缺氧处理 ,半定量RT PCR ,Westernblot法分别检测缺氧 2、6、1 2和 2 4hSTAT 3mRNA和酪氨酸活性水平的变化 ;3 H TdR掺入法观察缺氧 6、1 2和 2 4h细胞增殖情况。结果 缺氧 2hSTAT 3mRNA表达升高 ,6h达高峰 ,1 2h开始下降 ,但仍高于常氧组 ;Westernblot定量分析示缺氧 6hSTAT 3酪氨酸磷酸化水平升高 ,1 2h达高峰 ,2 4h下降 ;3 H TdR掺入法测定结果示PASMCs3 H TdR掺入量在缺氧 6h开始增加 ,随着缺氧时间延长 ,3 H TdR掺入量逐渐增加 ,至缺氧 2 4h3 H TdR掺入量最高。结论缺氧诱导PASMCs中STAT 3mRNA和蛋白酪氨酸活性水平增加 ,提示STAT 3在缺氧致PASMCs增殖过程中可能发挥重要作用
Objective To investigate the effects of hypoxia on the expression of signal transducer and activator of transcription 3 (STAT3) and cell proliferation in pulmonary artery smooth muscle cells (PASMCs) of rats. Methods Primary cultured PASMCs were treated with tissue block method and subjected to hypoxia. Semi-quantitative RT-PCR and Western blotting were used to detect the changes of STAT 3 mRNA and tyrosine activity in hypoxia, respectively. The incorporation of 3 H TdR Law observed hypoxia 6,1 2 and 2 4h cell proliferation. Results The expression of STAT 3 mRNA increased at 2 h and peaked at 6 h, but decreased at 12 h but still higher than that in normoxia group. Western blot analysis showed that the level of tyrosine phosphorylation increased 6 h after hypoxia, peaked at 12 h and reached the peak at 24 h 3 H TdR incorporation assay showed that 3 H TdR incorporation in PASMCs increased from 6 h after hypoxia. With the prolongation of hypoxia, the incorporation of 3 H TdR increased gradually, highest. Conclusion The increase of STAT3 mRNA and protein tyrosine activity in hypoxia-induced PASMCs suggests that STAT3 may play an important role in the process of hypoxia-induced PASMCs proliferation