高体积分数氧对人肺腺癌细胞硫氧还蛋白-1及硫氧还蛋白还原酶-1表达的影响  

Effects of Hyperoxia on Expressions of Thioredoxin-1 and Thioredoxin Reductase-1 in Human Adenocarcinoma Cells

作　　者：单瑞艳[1] 常立文[1] 李文斌[1] 刘伟[1] 陈燕[1] 曾凌空[1] 徐洪涛[1] 周玉容[1] 张佳[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院儿科,武汉430030

出　　处：《实用儿科临床杂志》2010年第20期1573-1575,共3页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(30770944)

摘　　要：目的研究高体积分数氧(高氧)对人肺腺癌A549细胞硫氧还蛋白-1(Trx1)及硫氧还蛋白还原酶-1(TrxR1)表达的影响,探讨Trx1及TrxR1在高氧肺损伤中的作用。方法体外传代培养A549细胞,待其生长至融合状态时随机分为高氧组和空气组。高氧组置于一饱和湿度密封箱中,通以950 mL.L-1氧气(O2)[含50 mL.L-1二氧化碳(CO2)]15 min;空气组置于同一室空气中15 min;2组同时置于37℃含50 mL.L-1CO2细胞培养箱中,分别于6 h、12 h、24 h提取其A549细胞总RNA和总蛋白。采取半定量反转录(RT)-PCR测定其Trx1 mRNA、TrxR1 mRNA的表达;采用Western blot检测其A549细胞Trx1蛋白的表达。结果 1.与空气组比较,A549细胞高氧暴露6 h、12 h时Trx1 mRNA表达显著增强(P=0.012,0.002),24 h时减弱,与空气组比较差异无统计学意义(P=0.795);与空气组比较,高氧暴露6 h TrxR1 mRNA无明显变化(P=0.878);12 h、24 h时TrxR1 mRNA表达明显增强(P=0.003,0.001)。2.与空气组比较,高氧暴露6 h Trx1蛋白有所增加但差异无统计学意义(P=0.575);12 h、24 h时Trx1蛋白表达均显著增强(P=0.003,0.026)。结论高氧暴露可上调A549细胞中Trx1及TrxR1的表达,提示内源性的Trx1及TrxR1在高氧肺损伤机制中提供了保护作用。Objective To investigate the expressions of thioredoxin-1（Trx1） and thioredoxin reductase-1（TrxR1） in human adenocarcinoma of lung A549 cells exposed to high concentration of oxygen,and explore the roles of Trx1 and TrxR1 in hyperoxic lung injury.Methods A549 cells were passaged and cultured in 50 mL·L-1 CO2 culture chamber,when approaching to the condition of subconfluence in CO2 culture chamber,the cells were randomly divided into air group and hyperoxia group.The cells in hyperoxia group were transferred to a sealed,humidified chamber which were flushed with 950 mL·L-1 O2/50 mL·L-1 CO2 for 15 min,while those in air group were placed in the air of the same room.Both of them were incubated at 37 ℃ in humidified air containing 50 mL·L-1 CO2 at the same time.After exposure to oxygen or air for 6 h,12 h and 24 h,the total RNA of A549 cells were isolated and cells protein were extracted.Trx1 mRNA and TrxR1 mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction.Western blot was used to detect the expression of Trx1 protein in A549 cells.Results 1.Compared with air group,the expressions of Trx1 mRNA were significantly increased in A549 cells exposed to hyperoxia for 6 h and 12 h（P=0.012,0.002）,and its expression became weak for 24 h and the difference was not significant between the both groups（P=0.795）.TrxR1 mRNA after hyperoxia exposure for 6 h had no significant change（P=0.878）;the expressions of TrxR1 mRNA were increased significantly exposed to hyperoxia for 12 h and 24 h（P=0.003,0.001）.2.Western blot showed,compared with air group,Trx1 protein expression after exposure to hyperoxia for 6 h was elevated,but the difference was not significant（P=0.575）.In contrast,the expressions of Trx1 protein were remarkably increased in A549 cells after exposure to hyperoxia for 12 h and 24 h compared to air group（P=0.003,0.026）.Conclusions After exposure to hypero-xia the expressions of Trx1 and TrxR1 can be significantly up-regulate in A549 cells,which ind

关 键 词：高体积分数氧 A549细胞 肺损伤 硫氧还蛋白-1 硫氧还蛋白还原酶-1 

分 类 号：R734.2[医药卫生—肿瘤] TB331[医药卫生—临床医学]