血红素氧合酶1对肺上皮细胞自噬和凋亡的影响及机制

摘要

目的 探讨血红素氧合酶1(HO-1)对肺泡上皮细胞自噬和凋亡的影响及机制.方法 采用前瞻性研究体外培养人源肺泡Ⅱ型上皮细胞A549,利用体外设计合成的小分子RNA(siRNA)沉默A549细胞HO-1基因的表达.实验分为:对照组(Control组)、siRNA-Con空载组(Con组)、HO-1 siRNA沉默组(siRNA组).Control组不作任何处理,Con组和HO-1 siRNA组分别转染siRNA-Con和HO-1 siRNA 48 h后取样本检测.采用荧光定量PCR(QT PCR)检测HO-1 mRNA水平;采用Western Blot法检测 HO-1、磷脂酰肌醇-3激酶(PI3K)、微管相关蛋白轻链3B(LC-3B)蛋白的表达变化;采用免疫荧光结合荧光显微镜观察各组细胞内HO-1和LC3B表达定位;采用JC-1结合流式细胞仪检测细胞线粒体膜电位情况.结果 与Control组相比,Con组HO-1、LC3B、PI3K蛋白表达,JC-1单体所占比率差异无统计意义(P >0.05);与Con组相比,HO-1 siRNA组细胞HO-1、LC-3B、PI3K蛋白表达明显下降(P 0.05),the expression of HO-1,LC-3B and PI3K protein in siRNA group were significantly lower than those in Con group(P <0.05). The percentage of JC-1 monomer in HO-1 siRNA group increased and the number of early apoptotic cells increased(P <0.01). Conclusion Heme Oxygenase 1 may regulate the role of autophagy and apoptotic levels in lung epithelial cell A549 cells and its mechanism may be related to PI3K signaling pathway.