蟛蜞菊内酯通过调控miR-142-3p表达对肺炎链球菌感染的肺泡上皮细胞损伤的保护作用及其机制研究  

The protective effect and mechanism of wedelolactone on alveolar epithelial cell injury infected by Streptococcus pneumoniae by regulating the expression of miR-142-3p

作　　者：郑大炜[1] 代岩[1] 郭娜[1] 党强[1] 路尧 张鹏[1] 孙辉[1] 王春侠[1] ZHENG Da-wei;DAI Yan;GUO Na(Department of Respiratory and Critical Care Medicine,Nanyang City Center Hospital,Nanyang 473000,China)

机构地区：[1]河南省南阳市中心医院呼吸与危重一科,河南南阳473000

出　　处：《中国实验诊断学》2021年第12期1831-1836,共6页Chinese Journal of Laboratory Diagnosis

摘　　要：目的探讨蟛蜞菊内酯(WEL)对肺炎链球菌感染的肺泡上皮细胞损伤的作用及分子机制。方法将肺泡上皮细胞A549分为对照组、感染组、感染组+蟛蜞菊内酯低、中、高浓度(WEL-L、M、H)组、感染组+miR-NC组、感染组+miR-142-3p组、感染组+WEL+anti-miR-NC组、感染组+WEL+anti-miR-142-3p组。酶联免疫吸附法(ELISA)检测IL-6、IL-10水平;流式细胞术检测细胞凋亡;蛋白质印迹(Western blot)法检测蛋白表达;实时荧光定量PCR(RT-qPCR)检测miR-142-3p表达水平。结果肺炎链球菌感染的肺泡上皮细胞中IL-10水平降低,IL-6水平升高,细胞凋亡率升高,Bcl-2表达水平降低,Bax表达水平升高,miR-142-3p表达水平降低(P<0.05)。不同浓度蟛蜞菊内酯处理及过表达miR-142-3p后,IL-10水平升高,IL-6水平降低,细胞凋亡率降低,Bcl-2表达水平升高,Bax表达水平降低,miR-142-3p表达水平升高,且呈浓度依赖性(P<0.05)。抑制miR-142-3p表达逆转了蟛蜞菊内酯对肺炎链球菌感染的肺泡上皮细胞损伤的作用。结论蟛蜞菊内酯可能通过上调miR-142-3p对肺炎链球菌感染的肺泡上皮细胞损伤起保护作用。Objective To explore the effect and molecular mechanism of wedelolactone(WEL)on the damage of alveolar epithelial cells infected by Streptococcus pneumoniae.Methods Alveolar epithelial cells A549 were divided into control group,infection group,infection group + wedelolactone low,medium and high concentration(WEL-L,M,H)group,infection group+ miR-NC group,infection group+ miR-142-3 p group,infection group+WEL+anti-miR-NC group,infection group+WEL+anti-miR-142-3 p group.Enzyme-linked immunosorbent assay(ELISA)was used to detect IL-6 and IL-10 levels;flow cytometry was used to detect cell apoptosis;Western blot was used to detect protein expression;real-time fluorescent quantitative PCR(RT-qPCR)was used to detect miR-142-3 p expression.Results The level of IL-10 in the alveolar epithelial cells infected by Streptococcus pneumoniae was decreased,the level of IL-6 was increased,the rate of apoptosis was increased,the expression of Bcl-2 was decreased,the expression of Bax was increased,and the expression of miR-142-3 p was decreased(P<0.05).After treatment with different concentrations of wedelolactone and overexpression of miR-142-3 p,IL-10 level was increased,IL-6 level was decreased,apoptosis rate was decreased,Bcl-2 expression was increased,Bax expression was decreased,the expression of miR-142-3 p was increased,in a concentration-dependent manner(P<0.05).Inhibition of miR-142-3 p expression reversed the effect of wedelolactone on the damage of alveolar epithelial cells infected by Streptococcus pneumoniae.Conclusion Wedelolactone may protect the alveolar epithelial cells from streptococcus pneumoniae infection by up-regulating miR-142-3 p.

关 键 词：蟛蜞菊内酯 miR-142-3p 肺炎链球菌 肺泡上皮细胞 凋亡 炎症反应 

分 类 号：R563.1[医药卫生—呼吸系统]