基于NF-κB通路研究芍药苷对LPS诱导支气管上皮细胞炎症反应的作用及机制研究
Effect and mechanism of paeoniflorin on LPS-induced bronchial epithelial cell inflammatory response based on NF-κB pathway
何芳 1丁敏 1甄海宁 1陈亚隽 1薛欣欣1
作者信息
- 1. 武汉市第三医院呼吸与危重症医学科,武汉 430060
- 折叠
摘要
目的:探讨芍药苷通过调节NF-κB通路对脂多糖(LPS)诱导支气管上皮细胞炎症反应的作用.方法:体外培养BEAS-2B细胞进行芍药苷毒性试验和浓度筛选.将BEAS-2B细胞分成正常对照组、LPS组、LPS+CAPE组、LPS+PF组和LPS+CAPE+PF组.使用LPS(1 µg/ml)诱导BEAS-2B细胞炎症反应,按照不同分组给予芍药苷或NF-κB抑制剂CAPE干预,CCK-8检测细胞活力.流式细胞术检测细胞凋亡和细胞周期,ELISA检测IFN-γ、IL-4、IL-17C、IL-10的水平,Western blot检测p53、Bcl-2、Bax、Cyclin1、NF-κB和p-p65的蛋白表达水平.结果:芍药苷能增加细胞活力,抑制细胞凋亡,提高IFN-γ和IL-10水平(P<0.01),降低IL-4和IL-17C水平(P<0.01),下调p53、Bax、NF-κB和p-p65蛋白表达水平(P<0.01),上调Bcl-2和Cyclin1蛋白表达水平(P<0.01).NF-κB抑制剂CAPE干预后芍药苷作用效果更显著.结论:芍药苷可通过调控NF-κB通路降低LPS诱导的支气管上皮细胞炎症因子水平,从而抑制哮喘炎症反应.
Abstract
Objective:To explore the effect of paeoniflorin on LPS-induced bronchial epithelial cell inflammatory response by regulating NF-κB pathway.Methods:BEAS-2B cells were cultured in vitro for paeoniflorin toxicity assay and concentration screening.BEAS-2B cells were divided into control group,LPS group,LPS+CAPE group,LPS+PF group and LPS+CAPE+PF group.Inflammatory responses were induced in BEAS-2B cells using LPS(1 µg/ml),and cell viability was detected by CCK-8 assay after administration of paeoniflorin or CAPE interventions.Apoptosis and cell cycle were detected by flow cytometry.The levels of IFN-γ,IL-4,IL-17C and IL-10 were detected by ELISA.The protein expression levels of p53,Bcl-2,Bax,Cyclin1,NF-κB and p-p65 were detected by Western blot.Results:Paeoniflorin increased cell viability,inhibited apoptosis,increased IFN-γ and IL-10 levels(P<0.01),de-creased IL-4 and IL-17C levels(P<0.01),down-regulated the protein expression levels of p53,Bax,NF-κB and p-p65(P<0.01),and up-regulated the protein expression levels of Bcl-2 and Cyclin1(P<0.01).The effect of paeoniflorin was more significant after the intervention of NF-κB inhibitor CAPE.Conclusion:Paeoniflorin reduces LPS-induced inflammatory factor levels in bronchial epithelial cells by regulating the NF-κB pathway,thereby suppressing the asthmatic inflammatory response.
关键词
芍药苷/哮喘/炎症反应/支气管上皮细胞/NF-κB通路Key words
Paeoniflorin/Asthma/Inflammatory response/Bronchial epithelial cells/NF-κB pathway引用本文复制引用
基金项目
武汉市卫健委2020年中医药类及中西医结合类一般项目(WZ20C20)
出版年
2024
国家科技期刊平台
NSTL
万方数据