TIMP1基因对肺炎链球菌诱导的肺泡上皮细胞HEPApiC凋亡的影响及机制研究  

Effect of TIMP1 gene on apoptosis of HEPApiC induced by streptococcus pneumoniae and its mechanism

作　　者：常大芸 刘学工[1] 李宁华[1] 王晓丽[1] 戴蕾莲[1] 王晓辉[2] 王义涛[2] CHANG Da-yun;LIU Xue-gong;LI Ning-hua;WANG Xiao-li;DAI Lei-lian;WANG Xiao-hui;WANG Yi-tao(Department of Pediatrics,Weihai Municipal Hospital,Weihai,Shandong 264200,China;Clinical Laboratory,Weihai Municipal Hospital,Weihai,Shandong 264200,China)

机构地区：[1]威海市立医院儿科,山东威海264200 [2]威海市立医院检验科,山东威海264200

出　　处：《临床肺科杂志》2020年第1期40-45,共6页Journal of Clinical Pulmonary Medicine

摘　　要：目的探讨基质金属蛋白酶抑制剂1(TIMP1)基因对肺炎链球菌诱导的人肺泡上皮细胞HEPApiC凋亡的影响及其可能的机制。方法体外培养人肺泡上皮细胞HEPApiC,采用肺炎链球菌感染HEPApiC细胞,实时荧光定量PCR(QPCR)和Western blot检测细胞中TIMP1的表达水平。将HEPApiC细胞分为4组,空白对照组(未做任何处理的细胞)、感染组(肺炎链球菌感染的细胞)、阴性对照组(转染siRNA control+肺炎链球菌感染的细胞)和干扰组(转染TIMP1 siRNA+肺炎链球菌感染的细胞)。CCK-8法检测各组HEPApiC细胞活力,流式细胞术检测各组HEPApiC细胞凋亡情况,Western blot检测细胞中B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平,ELISA检测上清中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的含量。结果肺炎链球菌感染后HEPApiC细胞中TIMP1 mRNA和蛋白表达水平均明显上调。转染TIMP1 siRNA后HEPApiC细胞中TIMP1 mRNA和蛋白表达水平均明显下调。感染组细胞凋亡率、Bax表达水平、TNF-α、IL-1β和IL-6含量均明显高于空白对照组(P<0.05),而细胞活力和Bcl-2表达水平低于空白对照组(P<0.05);干扰组细胞凋亡率、Bax表达水平、TNF-α、IL-1β和IL-6含量均明显低于感染组和阴性对照组(P<0.05),而细胞活力和Bcl-2表达水平高于感染组和阴性对照组(P<0.05)。结论敲低TIMP1基因表达能够抑制肺炎链球菌诱导的HEPApiC细胞凋亡,其作用机制可能与抑制炎性因子TNF-α、IL-1β和IL-6的表达有关。Objective To investigate the effect of matrix metalloproteinase inhibitor 1(TIMP1)gene on the apoptosis of human alveolar epithelial cells HEPApiC induced by Streptococcus pneumoniae and its possible mechanism.Methods Human alveolar epithelial cells HEPApiC were cultured in vitro,HEPApiC cells were infected with streptococcus pneumoniae,and the expression level of TIMP1 was detected by QPCR and Western blot.The HEPApiC cells were divided into 4 groups.The blank control group,the infection group,the negative control group and the interference group were tested for the activity of HEPApiC cells by CCK-8 method.Flow cytometry was used to detect the apoptosis of HEPApiC cells in each group.Western blot analysis was used to detect Bcl-2 and Bax expression levels in cells.The content of TNF-α,IL-1βand IL-6 in supernatant was determined by ELISA.Results The expression levels of TIMP1 mRNA and protein in HEPApiC cells were significantly up-regulated after S.pneumoniae infection.The expression levels of TIMP1 mRNA and protein in HEPApiC cells were significantly down-regulated after transfection of TIMP1 siRNA.The apoptotic rate,Bax expression,TNF-α,IL-1βand IL-6 levels in the infected group were significantly higher than those in the blank control group(P<0.05),while the cell viability and Bcl-2 expression levels were lower than the blank control group(P<0.05).Apoptosis rate,Bax expression level,TNF-α,IL-1βand IL-6 levels in the interference group were significantly lower than those in the infected group and the negative control group(P<0.05),while cell viability and Bcl-2 were higher than those of the infected group and the negative control group(P<0.05).Conclusion Knockdown of TIMP1 gene expression can inhibit the apoptosis of HEPApiC cells induced by streptococcus pneumoniae,and its mechanism may be related to the inhibition of the expression of inflammatory factors TNF-α,IL-1βand IL-6.

关 键 词：TIMP1基因 肺泡上皮细胞 凋亡 肺炎 

分 类 号：R73[医药卫生—肿瘤]