香烟烟雾提取物刺激下人支气管上皮细胞内质网应激对线粒体融合功能的影响  

CSE induced ERS in NHBE disturbs mitochondrial fusion

作　　者：龚丽[1] 罗百灵[1] 宋超[1] 

机构地区：[1]中南大学湘雅医院呼吸内科,长沙410008

出　　处：《国际呼吸杂志》2017年第9期688-692,共5页International Journal of Respiration

摘　　要：目的探讨在香烟烟雾提取物（cigarette smoking extract，CSE）作用下人支气管上皮细胞（NHBE）中内质网应激对线粒体融合功能的影响。方法体外培养NHBE，用CCK8法检测CSE不同浓度及不同时间干预对细胞存活率的影响，确定CSE干预的最佳浓度及时间。实验分为4组：对照组，内质网应激抑制剂（4-PBA）组，CSE干预组，4PBA＋CSE组，并分别给予相应浓度CSE及4-PBA干预。qPCR及Westernblot法检测内质网应激特异性蛋白CHOP及线粒体融合蛋白2（MFN2）的表达。结果细胞的生存率随CSE干预浓度及时间的增加而下降，CSE最佳干预浓度为5％，时间为12h。与正常对照组比较，CSE干预组CHOP表达增加，MFN2表达下降，差异有统计学意义[-mRNA水平上均值差（I-J）分别为4．17，0．67；蛋白水平上均．X差分别为3．38，1．26；P值均〈0．05]；4-PBA＋CSE组与CSE干预组比较，CHOP表达下降，MFN2表达增加，差异有统计学意义[mRNA水平上均值差（I-J）分别为2．47，0．41；蛋白水平上均值差分别为1．90，0．85，P值均〈0．05]。结论CSE作用下NHBE发生内质网应激，内质网应激通过减少MFN2影响线粒体融合。Objective To investigate the effect of cigarette smoking extract（CSE） induced ERS on mitochondrial fussion in NHBE. Methods NHBE were cultured in vitro and were treated with CSE of different concentration for different time. CCK8 was used to test the most suitable concentration and time for CSE to stimulate NHBE. Then the experiment was divided into four groups： normal control group, 4-PBA group, CSE group and 4-PBA-7 CSE group. Different group was processed with corresponding treatment. QPCR and western blot were used to detect mRNA and protein expression of CHOP and MFN2.Results Survival rate of CSE treated cells was negatively correlated with CSE concentration and time. The most suitable concentration and time of CSE to stimulate NHBE was 5 % and 12 h respectively. Compared with the control group, both mRNA and protein expression of CHOP in the CSE group were increased （the mean difference is 4.17,3.38 respectively, both P 〈0.05 ） ; however, mRNA and protein expression of MFN2 in the CSE group were decreased compared to those of the control group（the mean difference is 0.67,1.26 respectively, both P〈0.05）. The similarly results were found when CSE group compared with 4-PBA-7 CSE group. Conclusions CSE can result in ERS and decreased expression of MFN2 in NHBE. ERS disturbs mitochondrial fusion via MFN2.

关 键 词：肺疾病 阻塞性 内质网应激 线粒体 CCAAT/增强子结合蛋白 线粒体融合蛋白2 

分 类 号：R563.9[医药卫生—呼吸系统]