小干扰RNA干扰高氧暴露A549细胞Nrf2对谷胱甘肽S转移酶及白细胞介素1β的影响  

Effects of siRNA interference with Nrf2 on the regulation of glutathione S-transferase and IL-1βin A549 cells exposed to hyperoxia

作　　者：蔡怡子 李爱国 朱莹莹 张潇月 龚小慧[1] 蔡成[1] Cai Yizi;Li Aiguo;Zhu Yingying;Zhang Xiaoyue;Gong Xiaohui;Cai Cheng(Department of Neonatology,Shanghai Children's Hospital,Shanghai Jiao Tong University,Shanghai 200062,China;Department of Pediatrics,Tongren Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200336,China)

机构地区：[1]上海市儿童医院,上海交通大学附属儿童医院新生儿科,200062 [2]上海交通大学医学院附属同仁医院儿科,200336

出　　处：《中华新生儿科杂志（中英文）》2021年第5期58-62,共5页Chinese Journal of Neonatology

基　　金：国家自然科学基金面上项目(81571467)。

摘　　要：目的探讨高氧暴露下A549细胞中小干扰RNA(short interference RNA,siRNA)干扰红系衍生核因子相关因子2(nuclear factor erythroid-2-related factor 2,Nrf2)后,Nrf2与谷胱甘肽S转移酶(glutathione-S-transferase,GST)、白细胞介素1β(interleukin-1β,IL-1β)的基因表达变化及细胞凋亡情况。方法将体外培养的正常A549细胞株根据是否使用siRNA干扰、是否暴露于高氧环境(95%O2+5%CO2),分为常氧实验组、常氧对照组、高氧实验组、高氧对照组。采用实时定量聚合酶链反应检测各组细胞中Nrf2、GST和IL-1β的mRNA表达水平;采用流式细胞术检测高氧对照组和高氧实验组在不同时间点细胞凋亡情况;使用方差分析检验各组之间基因相对表达及A549细胞凋亡情况。结果(1)与常氧对照组相比,高氧对照组A549细胞Nrf2、GST基因表达升高、IL-1β基因表达下降(P<0.05),常氧实验组Nrf2、GST基因表达下降、IL-1β基因表达升高(P<0.05)。(2)常氧实验组和高氧实验组Nrf2、GST、IL-1β基因表达水平比较差异无统计学意义(P>0.05)。(3)与高氧对照组相比,高氧实验组Nrf2、GST基因表达下降、IL-1β基因表达升高(P<0.05)。(4)与高氧对照组相比,高氧实验组A549细胞凋亡比例在24 h、48 h及72 h增高(P<0.05)。结论siRNA干扰Nrf2后,通过调控GST和IL-1β的表达抵抗氧化应激,减轻炎症反应、抑制细胞凋亡。Objective To study the gene expression of nuclear factor erythroid-2-related factor 2(Nrf2),glutathione-S-transferase(GST)and interleukin-1β(IL-1β)in A549 cells exposed to hyperoxia and cell apoptosis after siRNA interference with Nrf2.Method Normal A549 cell lines were assigned into normoxia+siRNA group,normoxia+control group,hyperoxia+siRNA group and hyperoxia+control group according to whether siRNA interference was used and the exposure environment(normoxia/hyperoxia).The hyperoxia environment contained 95%O2 and 5%CO2.The levels of mRNA expression of Nrf2,GST and IL-1βwere detected using quantitative real-time polymerase chain reaction(qRT-PCR).Flow cytometry was used to examine cell apoptosis of the hyperoxia+control group and hyperoxia+siRNA group at different time points.Analysis of variance(ANOVA)was used to test the relative gene expression and apoptosis of A549 cells.Result(1)Compared with the normoxia+control group,the expression of Nrf2 and GST in the hyperoxia+control group was significantly increased(P<0.05),and the expression of IL-1βwas significantly decreased(P<0.05);the expression of Nrf2 and GST in the normoxia+siRNA group decreased significantly(P<0.05),while the expression of IL-1βincreased significantly(P<0.05).(2)Compared with the normoxia+siRNA group,Nrf2 expression in the hyperoxia+siRNA group showed no significant changes(P=0.230),GST expression increased slightly(P=0.057),and IL-1βexpression decreased slightly(P=0.112).(3)Compared with the hyperoxia+control group,the expression of Nrf2 and GST in the hyperoxia+siRNA group decreased significantly(P<0.05),and the expression of IL-1βincreased significantly(P=0.042).(4)Compared with the hyperoxia+control group,the apoptosis of A549 cells in the hyperoxia+siRNA group increased significantly at 24 h,48 h and 72 h(P<0.05).Conclusion After interfering with Nrf2,siRNA may regulate the expression of GST and IL-1β,preventing oxidative stress,reducing inflammatory response and inhibiting apoptosis.

关 键 词：小干扰RNA 红系衍生核因子相关因子2 谷胱甘肽-S-转移酶 白细胞介素1Β 

分 类 号：R722.6[医药卫生—儿科]