阿尔泰金莲花总黄酮通过miR-23b-3p抑制RSV感染支气管上皮细胞凋亡和炎症反应的分子机制研究  

Molecular Mechanism of Ttrollius altaicus Flavonoids Inhibiting Apoptosis and Inflammation Response of Bronchial Epithelial Cells Infected with RSV through miR-23b-3p

作　　者：尚耀民[1] 邢宝春 胡旭 周晓蕾[1] 梁瑞霞[1] 张一[2] SHANG Yaomin;XING Baochun;HU Xu;ZHOU Xiaolei;LIANG Ruixia;ZHANG Yi(Henan Provincial Chest Hospital,Zhengzhou 450000,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南省胸科医院,河南郑州450000 [2]河南中医药大学第一附属医院,河南郑州450000

出　　处：《中医药信息》2022年第9期33-38,共6页Information on Traditional Chinese Medicine

基　　金：河南省医学科技攻关计划项目(LHGJ20210230)。

摘　　要：目的:探讨阿尔泰金莲花总黄酮对呼吸道合胞病毒(RSV)感染支气管上皮细胞凋亡和炎症反应的影响及分子机制。方法:将人支气管上皮细胞16-HBE分为NC组、RSV组、RSV+低、中、高剂量组、RSV+miR-23b-3p组、RSV+miR-NC组、anti-miR-23b-3p+RSV+高剂量组和anti-miR-NC+RSV+高剂量组。细胞计数(CCK-8)试剂检测细胞活性,流式细胞术检测细胞凋亡情况,Westernblot法检测蛋白表达情况,ELISA法检测TNF-α、IL-6、IL-1α水平,实时荧光定量PCR(RT-qPCR)检测miR-23b-3p表达水平。结果:RSV感染的16-HBE细胞活性、CyclinD1表达水平降低,凋亡率、CleavedCaspase-3、Bax表达水平、TNF-α、IL-6、IL-1α水平升高,miR-23b-3p表达水平降低(P<0.01)。不同剂量阿尔泰金莲花总黄酮处理后,16-HBE细胞活性、CyclinD1表达水平升高,凋亡率、CleavedCaspase-3、Bax表达水平、TNF-α、IL-6、IL-1α水平降低,miR-23b-3p表达水平升高(P<0.01)。过表达miR-23b-3p后,细胞活性、CyclinD1表达水平升高,凋亡率、CleavedCaspase-3、Bax表达水平、TNF-α、IL-6、IL-1α水平降低(P<0.01)。抑制miR-23b-3p表达可以逆转阿尔泰金莲花总黄酮对RSV感染的16-HBE细胞活性,凋亡和炎症因子水平的影响。结论:阿尔泰金莲花总黄酮可能通过上调miR-23b-3p表达抑制RSV感染的支气管上皮细胞凋亡和炎症反应。Objective:To explore the effect and the molecular mechanism of Ttrollius altaicus flavonoids on apoptosis and inflammatory response of bronchial epithelial cells infected with respiratory syncytial virus(RSV).Methods:Human bronchial epithelial cells 16-HBE were divided into NC group,RSV group,RSV+Ttrollius altaicus flavonoids groups of low-dose,medium-dose and high-dose,RSV+miR-23b-3p group,RSV+miR-NC group,anti-miR-23b-3p+RSV+high-dose group,anti-miR-NC+RSV+high-dose group.Cell viability was detected by CCK-8 kit;cell apoptosis rate was detected by flow cytometry;protein expression was detected by Western blot;the levels of TNF-α,IL-6 and IL-1αwere measured by ELISA;miR-23b-3p expression was detected by RT-qPCR.Results:Cell activity and CyclinD1 expression of RSV-infected 16-HBE were decreased(P<0.01);apoptosis rate,expressions of Cleaved Caspase-3 and Bax,and the levels of TNF-α,IL-6 and IL-1αwere increased(P<0.01);miR-23b-3p expression was decreased(P<0.01).After the intervention of Trollius altaicus flavonoids with different doses,16-HBE cell activity and CyclinD1 expression were increased(P<0.01);apoptosis rate,expressions of Cleaved Caspase-3 and Bax,as well as the levels of TNF-α,IL-6 and IL-1αwere decreased(P<0.01);the expression of miR-23b-3p was increased(P<0.01).After over-expression of miR-23b-3p,16-HBE cell activity and CyclinD1 expression were increased(P<0.01);apoptosis rate,expressions of Cleaved Caspase-3 and Bax,as well as the levels of TNF-α,IL-6 and IL-1αwere decreased(P<0.01).Inhibiting the expression of miR-23b-3p could reverse the effect of Trollius altaicus flavonoids on the activity,apoptosis and inflammatory factor levels of RSV-infected 16-HBE cells.Conclusion:Ttrollius altaicus flavonoids may inhibit RSV-infected bronchial epithelial cell apoptosis and inflammation response by up-regulating the expression of miR-23b-3p.

关 键 词：阿尔泰金莲花总黄酮 miR-23b-3p 支气管上皮细胞 凋亡 炎症反应 

分 类 号：R285[医药卫生—中药学]