Circ_0038467靶向miR-182调控肺炎链球菌(Sp)诱导的人肺泡上皮细胞系HPAEpiC损伤

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (9): 1277-1284.

Circ_0038467靶向miR-182调控肺炎链球菌(Sp)诱导的人肺泡上皮细胞系HPAEpiC损伤

刘青¹, 张蕾², 张利元³, 王世凤⁴, 左立旻^1*

电子科技大学医学院附属妇女儿童医院成都市妇女儿童中心医院 1.急诊科; 2.呼吸科; 3.检验科; 4.病理科, 四川成都 611731

收稿日期:2020-09-22 修回日期:2021-02-02 出版日期:2021-09-05 发布日期:2021-09-02
通讯作者: *fob6er@163.com
基金资助:
四川省卫生和计划生育委员会科研课题(16PJ067)

Circ_0038467 targets at miR-182 for regulating S. pneumoniae (Sp)-induced injury of human alveolar epithelial cell line HPAEpiC

LIU Qing¹, ZHANG Lei², ZHANG Li-yuan³, WANG Shi-feng⁴, ZUO Li-min^1*

1. Department of Emergency; 2. Department of Respiratory Medicine; 3. Clinical Laboratory; 4. Department of Pathology, Chengdu Women's and Children's Central Hospital, School of Medicine, University of Electronic Science and Techology of China, Chengdu 611731, China

Received:2020-09-22 Revised:2021-02-02 Online:2021-09-05 Published:2021-09-02
Contact: *fob6er@163.com

摘要/Abstract

摘要： 目的探讨环状RNA 0038467(circ_0038467)靶向miR-182对肺炎链球菌(Sp)诱导的肺泡上皮细胞系HPAEpiC凋亡和内质网应激的影响。方法将HPAEpiC细胞分为对照(NC)组(未做任何处理)、Sp组(Sp感染)、Sp+si-NC组(转染si-NC后Sp感染)、Sp+si-circ_0038467组(转染si-circ_0038467后Sp感染)、Sp+miR-NC组(转染miR-NC后Sp感染)、Sp+miR-182组(转染miR-182 mimics后Sp感染)、Sp+si-circ_0038467+anti-miR-NC组(转染si-circ_0038467和anti-miR-NC后Sp感染)、Sp+si-circ_0038467 +anti-miR-182组(转染si-circ_0038467和anti-miR-182后Sp感染)。实时定量PCR检测细胞中circ_0038467和miR-182表达量;流式细胞测量术检测细胞凋亡率;蛋白印迹法检测B细胞淋巴瘤(Bcl-2)、重链结合蛋白(BIP)、Bcl相关X蛋白(Bax)和CCAAT增强子结合蛋白同源蛋白(CHOP)、半胱氨酸蛋白酶3(caspase-3)和裂解的caspase-3(cleaved-caspase-3)蛋白表达量。结果 Sp感染后HPAEpiC细胞凋亡率、circ_0038467、Bax、caspase-3、cleaved-caspase-3、BIP和CHOP表达显著升高(P＜0.05),而Bcl-2表达显著降低(P＜0.05)。抑制circ_0038467或过表达miR-182后Sp诱导的HPAEpiC凋亡率、circ_0038467、Bax、caspase-3、cleaved-caspase-3、BIP和CHOP表达显著降低(P＜0.05),而Bcl-2表达显著升高(P＜0.05)。抑制miR-182表达能够减弱circ_0038467抑制对Sp诱导的HPAEpiC凋亡以及内质网应激的影响(P＜0.05)。结论抑制circ_0038467可减弱Sp诱导的肺泡上皮细胞凋亡和内质网应激,其机制与靶向调控miR-182表达有关。

关键词: circ_0038467, 肺炎链球菌, 肺泡上皮细胞, 凋亡, 内质网应激

Abstract: Objective To investigate the effect of circular RNA 0038467 (circ_0038467) targeting at miR-182 on apoptosis and endoplasmic reticulum stress of alveolar epithelial cell line(HPAEpiC) induced by S. pneumoniae (Sp). Methods HPAEpiC cells were divided into control (NC) group (without any treatment), Sp group (Sp infection), Sp+si-NC group (Sp infection after transfection by si-NC), Sp+si-circ_0038467 group (Sp infection after transfection by si-circ_0038467), Sp+miR-NC group (Sp infection after transfection of miR-NC), and Sp+miR-182 group (Sp infection after transfection of miR-182 mimics), Sp+si-circ_0038467+anti-miR-NC group (Sp infection after transfection by si-circ_0038467 and anti-miR-NC), Sp+si-circ_0038467+anti-miR-182 group (Sp infection after transfection by si-circ_0038467 and anti-miR-182). The expressions of circ_0038467 and miR-182 were detected by RT-qPCR. Flow cytometry detected cell apoptosis; Western blot analyzed the expression level of B-cell lymphoma (Bcl-2), heavy chain binding protein (BIP), Bcl associated X protein (Bax) and CCAAT enhancer binding protein homologous protein (CHOP), cysteine protease 3 (caspase-3), and cleaved-caspase-3 proteins. The effects of circ_0038467 and miR-182 expression on HPAEpiC cell apoptosis and endoplasmic reticulum stress induced by S. pneumoniae were analyzed. Results The apoptosis rate of HPAEpiC cells, the expression of circ_0038467, Bax, caspase-3, cleaved-caspase-3, BIP and CHOP were significantly increased after Sp infection (P＜0.05), whereas the expression of Bcl-2 was significantly decreased(P＜0.05). After circ_0038467 inhibition or miR-182 over-expression, the apoptosis rate, expression of circ_0038467, Bax, caspase-3, cleaved-caspase-3, BIP and CHOP were significantly reduced (P＜0.05), whereas Bcl-2 expression was significantly increased(P＜0.05). miR-182 inhibition can attenuate the effect of circ_0038467 inhibition on Sp induced HPAEpiC apoptosis and endoplasmic reticulum stress (P＜0.05). Conclusions Inhibition of circ_0038467 can attenuate Sp induced alveolar epithelial cell apoptosis and endoplasmic reticulum stress, and its mechanism is related to the targeted regulation of miR-182 expression.

Key words: circ_0038467, S. pneumoniae, alveolar epithelial cells, apoptosis, endoplasmic reticulum stress

中图分类号:

Q932

刘青, 张蕾, 张利元, 王世凤, 左立旻. Circ_0038467靶向miR-182调控肺炎链球菌(Sp)诱导的人肺泡上皮细胞系HPAEpiC损伤[J]. 基础医学与临床, 2021, 41(9): 1277-1284.

LIU Qing, ZHANG Lei, ZHANG Li-yuan, WANG Shi-feng, ZUO Li-min. Circ_0038467 targets at miR-182 for regulating S. pneumoniae (Sp)-induced injury of human alveolar epithelial cell line HPAEpiC[J]. Basic & Clinical Medicine, 2021, 41(9): 1277-1284.

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