转染血红素加氧酶-1能抑制乙醇诱导的成骨细胞凋亡

Heme oxygenase-1 transfection can inhibit osteoblast apoptosis induced by ethanol

目的 观察转染血红素氧合酶-1（HO-1）能否抑制乙醇诱导的成骨细胞凋亡作用.方法 利用质粒将HO-1转染进入人成骨细胞,乙醇干预24h后,Hoechst染色观察成骨细胞核变化,流式细胞仪检测细胞凋亡,Western bolt检测细胞中凋亡蛋白酶活化因子1（APAF1）、半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3和细胞色素-C（Cyt-C）表达量.结果 Hoechst染色结果显示HO-1转染组少量细胞胞核发生改变,而阳性对照组中胞核发生改变的较多.流式细胞仪和Western blot检测结果显示,与阴性对照组比较,HO-1转染组和阳性对照组细胞凋亡率、APAF1、Caspase-3和Cyt-C表达水平均明显升高（P＜0.05）;而与阳性对照组比较,HO-1转染组细胞凋亡率、APAF1、Caspase-3和Cyt-C表达量均明显降低（P＜0.05）.结论 转染HO-1能抑制乙醇诱导的成骨细胞凋亡,这一作用可能与HO-1抑制细胞中APAF1、Caspase-3和Cyt-C的表达有关.

Objective To identify whether heme oxygenase-1 （HO-1） transfection can inhibit osteoblast apoptosis induced by ethanol.Methods HO-1 was transfected into osteoblasts using plasmid.After osteoblasts were exposed to ethanol for 24 h,the change of nucleus was observed by Hoechst stain.Apoptosis of osteoblasts was measured using flow cytometry.The levels of APAF1,cysteinyl aspartate-specific protease （Caspase）-3 and cytochrome-C （Cyt-C） were measured by Western blotting.Results After osteoblasts were exposed to ethanol for 24 h,there were a little of apoptosis cells in HO-1 transfection group,and there were a lot of apoptosis cells in positive control group.As compared with negative control group,the apoptosis rate and the levels of APAF1,caspase-3 and Cyt-C were all significantly increased in HO-1 transfection group and positive control group （P 〈0.05）.As compared with the positive control group,the apoptosis rate and the levels of APAF1,Caspase-3 and Cyt-C were all significantly decreased in HO-1 transfectian group （P 〈 0.05）.Conclusion HO-1 transfection can inhibit osteoblast apoptosis induced by ethanol,which was related to the inhibitory effects of HO-1 on APAF1,Caspase-3 and Cyt-C.