miR-29b通过调控Bax基因抑制LPS诱导的肺泡上皮细胞系A549凋亡

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (6): 797-802.

miR-29b通过调控Bax基因抑制LPS诱导的肺泡上皮细胞系A549凋亡

潘姝, 文丹宁^*, 李华东

武汉市金银潭医院感染科, 湖北武汉 430000

收稿日期:2019-06-21 修回日期:2019-11-06 出版日期:2020-06-05 发布日期:2020-05-29
通讯作者: *1696205723@qq.com
基金资助:
武汉市卫生局科研项目(WX15D49)

miR-29b inhibits LPS-induced apoptosis of alveolar epithelial cell line A549 through regulating Bax gene

PAN Shu, WEN Dan-ning^*, LI Hua-dong

Department of Infectious Diseases, Wuhan Jinyin Tan Hospital, Wuhan 430000, China

Received:2019-06-21 Revised:2019-11-06 Online:2020-06-05 Published:2020-05-29
Contact: *1696205723@qq.com

摘要/Abstract

摘要： 目的探讨miR-29b对脂多糖(LPS)诱导的肺泡上皮细胞系A549凋亡的影响及其机制。方法将体外培养的A549细胞分为对照组、LPS组(给予10 mg/L的LPS处理)、LPS+miR-NC组(转染miR-29b mimics 阴性对照后给予LPS处理)、LPS+miR-29b组(转染miR-29b mimics 后给予LPS处理);用RT-qPCR检测细胞中miR-29b的表达水平;MTT法检测细胞存活率;流式细胞仪检测细胞凋亡率;Western blot检测Bcl-2、Bax和cleaved caspase-3蛋白的表达;双荧光素酶报告基因实验检测Bax和miR-29b的靶向关系。结果与对照组相比,LPS组、LPS+miR-NC组和LPS+miR-29b组细胞中miR-29b、Bcl-2蛋白的表达水平和细胞存活率均明显降低,而细胞凋亡率和Bax、cleaved caspase-3蛋白的表达水平均明显升高(P＜0.05);与LPS组相比,LPS+miR-29b组细胞中miR-29b、Bcl-2蛋白的表达水平和细胞存活率均明显升高,而细胞凋亡率和Bax、cleaved caspase-3蛋白的表达水平均明显降低(P＜0.05)。双荧光素酶报告基因实验证实Bax是miR-29b的潜在靶基因。结论 miR-29b可抑制LPS诱导的A549细胞凋亡,其作用机制可能与靶向调控Bax表达有关。

关键词: 肺泡上皮细胞, miR-29b, LPS, 细胞凋亡, Bax

Abstract: Objective To investigate the effect of miR-29b on LPS-induced apoptosis of alveolar epithelial cell line A549 and its mechanism. Methods A549 cells were divided into control group, LPS group (treated with 10 mg/L LPS), LPS+miR-NC group (treated with LPS after transfection of miR-29b mimics negative control), LPS+miR-29b group(treated with LPS after transfection of miR-29b mimics), the expression level of miR-29b in each group was detected by RT-qPCR, cell viability was detected by MTT assay, and apoptosis rate was detected by flow cytometry.The expressions of Bcl-2, Bax and cleaved caspase-3 protein were detected by Western blot, and the dual luciferase reporter gene assay was used to detect the targeting relationship between Bax and miR-29b. Results Compared with the control group, the expression levels of miR-29b and Bcl-2 proteins and cell viability in the LPS group, LPS+miR-NC group and LPS+miR-29b group were significantly decreased, while the apoptosis rate was decreased. The expression of Bax and cleaved caspase-3 proteins significantly increased (P＜0.05). Compared with LPS group, the expression of miR-29b and Bcl-2 proteins and cell survival rate in LPS+miR-29b cells were significantly higher, while the apoptotic rate, the expression of Bax and cleaved caspase-3 protein were all significantly lower(P＜0.05). Dual luciferase reporter assays confirmed that Bax could be a potential target gene for miR-29b. Conclusions miR-29b can inhibit the apoptosis of alveolar epithelial cells A549 induced by LPS, and its mechanism may be related to the targeted regulation of Bax expression.

Key words: alveolar epithelial cells, miR-29b, LPS, apoptosis, Bax

中图分类号:

R966

潘姝, 文丹宁, 李华东. miR-29b通过调控Bax基因抑制LPS诱导的肺泡上皮细胞系A549凋亡[J]. 基础医学与临床, 2020, 40(6): 797-802.

PAN Shu, WEN Dan-ning, LI Hua-dong. miR-29b inhibits LPS-induced apoptosis of alveolar epithelial cell line A549 through regulating Bax gene[J]. Basic & Clinical Medicine, 2020, 40(6): 797-802.

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