摘要
目的探讨槲皮素对呼吸道合胞病毒(RSV)感染的支气管上皮细胞凋亡和炎症反应的影响及可能机制。方法体外培养支气管上皮细胞16-HBE,用不同剂量(1、10、100μmol/L)槲皮素干预RSV感染的16-HBE细胞24 h后,CCK-8法检测细胞活性,流式细胞术检测细胞凋亡,蛋白质印迹法检测细胞中Cleaved-caspase-3蛋白表达,试剂盒检测细胞培养上清液中TNF-α、IL-6和IL-1α表达量,q RT-PCR检测细胞中miR-140-5p表达。转染miR-140-5p模拟物或抑制剂至16-HBE细胞,上述相同方法观察miR-140-5p对RSV感染的16-HBE细胞凋亡及炎症反应的影响。结果与RSV组比较,槲皮素提高了RSV感染的16-HBE细胞活性(P<0.05),而降低了细胞凋亡率、细胞中Cleaved-caspase-3蛋白表达及培养上清液中TNF-α、IL-6和IL-1α水平(P<0.05)。槲皮素还促进了RSV感染的16-HBE细胞中miR-140-5p的表达。上调miR-140-5p提高了RSV感染的16-HBE细胞活性(P<0.05),而降低了细胞凋亡率、细胞中Cleaved-caspase-3蛋白表达及培养上清液中TNF-α、IL-6和IL-1α水平(P<0.05)。下调miR-140-5p降低了RSV感染的16-HBE细胞活性、凋亡及炎症因子TNF-α、IL-6和IL-1α表达的水平。结论槲皮素可抑制RSV感染的支气管上皮细胞凋亡和炎症反应,其作用机制可能与上调细胞中miR-140-5p表达有关。
This study was performed to investigate the effect of quercetin on the apoptosis and inflammation of bronchial epithelial cells infected with respiratory syncytial virus(RSV)and its possible mechanism.The bronchial epithelial cells 16-HBE were cultured in vitro.After different doses(1,10,100μmol/L)of quercetin were used to interfere with 16-HBE cells infected by respiratory syncytial virus for 24 h,the cell viability was detected by CCK-8method,and the cell apoptosis was detected by flow cytometry.The expression of Cleaved-caspase-3 protein in cells was detected by Western blotting;the expression of TNF-α,IL-6 and IL-1αin cell culture supernatant were detected by corresponding kits;and the expression of miR-140-5p in cells was detected by qRT-PCR.miR-140-5p mimics or inhibitors was transfected into 16-HBE cells,and then the same method described above were used to observe the effect of miR-140-5p on the apoptosis and inflammation of RSV-infected 16-HBE cells.Compared with the RSV group,quercetin increased the activity of RSV-infected 16-HBE cells(P<0.05),and decreased the apoptosis rate,Cleaved-caspase-3 protein expressionand the expression of TNF-α,IL-6 and IL-1α(P<0.05).Quercetin also promoted the expression of miR-140-5p in RSV-infected 16-HBE cells.Up-regulation of miR-140-5p could increase the activity of RSV-infected 16-HBE cells(P<0.05),and reduced the apoptosis rate,the expression of Cleaved-caspase-3 protein and theexpression of TNF-α,IL-6 and IL-1α(P<0.05).Down-regulation of miR-140-5p reduced the effect of quercetinon the activity,apoptosis and the expression of inflammatory factors TNF-α,IL-6 and IL-1αin RSV-infected 16-HBE cells.In conclusion,quercetin could inhibit RSV-infected bronchial epithelial cell apoptosis andinflammation,and the mechanism may be related to the up-regulation of miR-140-5p expression in the cells.
作者
章秋霞
陈炯科
吴伶莉
ZHANG Qiuxia;CHEN Jiongke;WU Lingli(Department of Laboratory,Second People's Hospital of Fuyang District,Hangzhou 311404,China;Department of Laboratory,Hangzhou Cancer Hospital,Hangzhou 310001,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2022年第10期883-889,共7页
Immunological Journal
