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首页|期刊导航|河北医学|甘草酸抑制呼吸道合胞病毒感染支气管上皮细胞凋亡的机制研究

甘草酸抑制呼吸道合胞病毒感染支气管上皮细胞凋亡的机制研究OACSTPCD

Mechanism of Inhibition of Apoptosis in Respiratory Syncytial Virus-Infected Bronchial Epithelial Cells by Glycyrrhizic Acid

中文摘要英文摘要

目的:探讨甘草酸对呼吸道合胞病毒(RSV)感染的支气管上皮细胞凋亡的影响,分析其机制是否与内源性酪氨酸激酶2(JAK2)/信号传导和转录激活因子 3(STAT3)信号通路有关.方法:将HBE细胞分为对照组、感染组(RSV组)、低剂量甘草酸组(RSV+L-甘草酸组)、中剂量甘草酸组(RSV+M-甘草酸组)、高剂量甘草酸组(RSV+H-甘草酸组)、RSV+Colivelin 组、RSV+H-甘草酸+SD-1029 组.对照组细胞不经任何处理;RSV组用含0.0001 PFU/mL的RSV病毒溶液培养细胞2h,再更换为正常培养液培养24h;RSV+L-甘草酸组、RSV+M-甘草酸组和RSV+H-甘草酸组在RSV感染HBE 细胞模型的基础上,用30、60、120μg/mL甘草酸处理的培养基培养的细胞;RSV+Colivelin 组在 RSV 感染 HBE 细胞模型的基础上,用含有浓度为0.5μmoL/L JAK2/STAT3 信号通路激活剂Colivelin 处理的培养基培养细胞;RSV+H-甘草酸+SD-1029 组在RSV感染HBE细胞模型的基础上,用120μg/mL甘草酸和10μmoL/L JAK2/STAT3 信号通路抑制剂 SD-1029 共同处理的培养基培养细胞.24h 后收集细胞,CCK-8 试剂盒检测细胞活性;流式细胞仪检测细胞凋亡;ELISA 检测细胞上清液中 IL-6、TNF-α 水平;western blot检测细胞中Bax、cleaved-Caspase-3、Bcl-2、p-JAK2、JAK2、p-STAT3、STAT3 蛋白表达情况.结果:与对照组比较,RSV组OD值、Bcl-2、cleaved-Caspase-3、p-JAK2/JAK2、p-STAT3/STAT3 蛋白表达均显著降低(P<0.05),细胞凋亡率、IL-6、TNF-α水平、Bax蛋白表达量显著升高(P<0.05);与RSV组比较,RSV+L-甘草酸组、RSV+M-甘草酸组、RSV+H-甘草酸组、RSV+Colivelin 组 OD 值、Bcl-2、cleaved-Caspase-3、p-JAK2/JAK2、p-STAT3/STAT3 蛋白表达均显著升高(P<0.05),细胞凋亡率、IL-6、TNF-α水平、Bax蛋白表达量显著降低(P<0.05);与RSV+H-甘草酸组比较,RSV+H-甘草酸+SD-1029 组OD值、Bcl-2、cleaved-Caspase-3、p-JAK2/JAK2、p-STAT3/STAT3 蛋白表达均显著降低(P<0.05),细胞凋亡率、IL-6、TNF-α水平、Bax蛋白表达量显著升高(P<0.05).结论:甘草酸可能通过激活 JAK2/STAT3 信号通路,促进RSV感染的支气管上皮细胞增殖,抑制RSV感染的支气管上皮细胞的凋亡.

Objective:To explore the effect of glycyrrhizic acid on the apoptosis of bronchial epithelial cells infected with respiratory syncytial virus(RSV)and to analyze whether the mechanism is related to the endogenous Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling path-way.Methods:Human bronchial epithelial(HBE)cells were divided into seven groups:control group,in-fection group(RSV group),low-dose glycyrrhizic acid group(RSV+L-glycyrrhizic acid group),medium-dose glycyrrhizic acid group(RSV+M-glycyrrhizic acid group),high-dose glycyrrhizic acid group(RSV+H-glycyrrhizic acid group),RSV+Colivelin group,and RSV+H-glycyrrhizic acid+SD-1029 group.The control group cells were untreated.The RSV group cells were cultured with RSV virus solution at 0.0001 PFU/mL for 2 hours and then switched to normal culture medium for 24 hours.The RSV+L-glycyrrhizic acid group,RSV+M-glycyrrhizic acid group,and RSV+H-glycyrrhizic acid group were cultured with media containing glycyr-rhizic acid at concentrations of 30,60,and 120μg/mL respectively after RSV infection.The RSV+Colivelin group cells were cultured with media containing 0.5μmoL/L of JAK2/STAT3 signaling pathway activator Col-ivelin after RSV infection.The RSV+H-glycyrrhizic acid+SD-1029 group cells were cultured with media con-taining 120μg/mL glycyrrhizic acid and 10μmoL/L JAK2/STAT3 signaling pathway inhibitor SD-1029 after RSV infection.After 24 hours,cells were collected.Cell viability was detected using the CCK-8 kit.Apopto-sis was detected by flow cytometry.Levels of IL-6 and TNF-α in cell supernatants were measured by ELISA.Protein expression of Bax,cleaved-Caspase-3,Bcl-2,p-JAK2,JAK2,p-STAT3,and STAT3 was ana-lyzed by western blot.Results:Compared to the control group,the RSV group showed significantly lower OD value,Bcl-2,cleaved-Caspase-3,p-JAK2/JAK2,and p-STAT3/STAT3 protein expression(P<0.05),and significantly higher cell apoptosis rate,IL-6,TNF-α levels,and Bax protein expression(P<0.05).Compared to the RSV group,the RSV+L-glycyrrhizic acid group,RSV+M-glycyrrhizic acid group,RSV+H-glycyrrhizic acid group,and RSV+Colivelin group showed significantly higher OD value,Bcl-2,cleaved-Caspase-3,p-JAK2/JAK2,and p-STAT3/STAT3 protein expression(P<0.05),and significantly lower cell apoptosis rate,IL-6,TNF-α levels,and Bax protein expression(P<0.05).Compared to the RSV+H-glycyrrhizic acid group,the RSV+H-glycyrrhizic acid+SD-1029 group showed significantly lower OD value,Bcl-2,cleaved-Caspase-3,p-JAK2/JAK2,and p-STAT3/STAT3 protein expression(P<0.05),and sig-nificantly higher cell apoptosis rate,IL-6,TNF-α levels,and Bax protein expression(P<0.05).Conclu-sion:Glycyrrhizic acid may promote the proliferation of bronchial epithelial cells infected with RSV and inhibit their apoptosis by activating the JAK2/STAT3 signaling pathway.

付晓康;梁薇薇;高帅;苏琴;尉全平;孙子梅

河北北方学院附属第二医院儿内,河北 张家口 075000河北省张家口市妇幼保健院儿内,河北 张家口 075000

呼吸道合胞病毒支气管上皮细胞甘草酸JAK2/STAT3信号通路凋亡

Respiratory syncytial virusBronchial epithelial cellsGlycyrrhetinic acidJAK2/STAT3 signaling pathwayApoptosis

《河北医学》 2024 (006)

912-918 / 7

2023年市级科技计划自筹经费项目,(编号:2322087D)

10.3969/j.issn.1006-6233.2024.06.06

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