siRNA沉默EGLN1基因对低氧下大鼠肺动脉平滑肌细胞生长的影响

Effects of EGLN1 siRNA on growth of rat pulmonary artery smooth muscle cells under hypoxic condition

目的:体外培养大鼠肺动脉平滑肌细胞(PASMCs),利用小干扰RNA技术转染PASMCs干扰EGLN1基因表达,检测细胞活力变化,从而验证EGLN1在PASMCs活力变化中的作用。方法:采用原代培养PASMCs,构建出特异的EGLN1 siRNA脂质体并转染到PASMCs;分别在常氧和低氧下进行细胞培养,采用Western blot检测PASMCs的EGLN1蛋白、VEGF蛋白表达水平;用CCK-8法检测细胞活力,探讨低氧条件下沉默EGLN1基因表达后对PASMCs活力的影响。结果:低氧下PASMCs活力变化和VEGF的蛋白水平表达较常氧下增加并呈时间依赖性;EGLN1沉默后,无论低氧和常氧下,PASMCs活力变化和VEGF的蛋白水平表达均受到抑制。结论:EGLN1基因参与调控低氧下大鼠PASMCs的生长,其调节可能是通过VEGF的介导而完成的。

AIM： To observe whether EGLN1 gene is involved in the growth of pulmonary arterial smooth muscle cells（ PASMCs） during hypoxia when EGLN1 gene expression was interference by siRNA. METHODS： The rat primary pulmonary arterial smooth muscle cells were cultured,and the specific lipidosome of EGLN1 siRNA was constructed and transfected into the PASMCs. The transfected PASMCs were cultured under hypoxia or normoxia conditions,respectively. The viability of the PASMCs was detected by CCK-8 assay. The protein expression of EGLN1 and vascular endothelial growth factor（ VEGF） was determined by Western blot. RESULTS： The viability of the PASMCs was increased and the protein expression of VEGF was up-regulated in the PASMCs under hypoxic condition in a time-dependent manner. In hypoxia or normoxia condition,the viability and VEGF protein expression of the PASMCs were suppressed by EGLN1 siRNA. CONCLUSION： EGLN1 gene may involve in the growth of rat PASMCs by regulating VEGF protein level under hypoxic condition.