摘要
目的探讨牡荆素对肺泡上皮细胞高迁移率族蛋白1(HMGB1)/晚期糖基化终产物受体(RAGE)通路及炎症损伤的影响。方法体外培养人肺泡上皮细胞HPAEpiC,分为对照组(不进行处理)、脂多糖组(10μg/mL脂多糖)、低浓度组(10μg/mL脂多糖+20μmol/L牡荆素)和高浓度组(10μg/mL脂多糖+60μmol/L牡荆素)。采用实时荧光定量PCR(qRT-PCR)法检测各组HPAEpiC细胞中HMGB1、RAGE mRNA表达情况;蛋白印迹(WB)法检测各组HPAEpiC细胞中HMGB1、RAGE蛋白表达情况;MTT法检测各组HPAEpiC细胞增殖情况;酶联免疫吸附(ELISA)法检测各组HPAEpiC细胞上清液中白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)水平。结果与对照组比较,脂多糖组HPAEpiC细胞中HMGB1、RAGE mRNA及蛋白表达水平显著升高(mRNA:t_(HMGB1)=14.269、t_(RAGE)=17.554,蛋白:t_(HMGB1)=19.200、t_(RAGE)=18.887,P均<0.05),细胞活力显著减弱(t=10.314,P<0.05),细胞上清液中IL-6、IL-8、TNF-α表达水平显著升高(t=45.731、64.827、46.063,P均<0.05);与脂多糖组比较,随着牡荆素的使用及浓度的升高,HPAEpiC细胞中HMGB1、RAGE mRNA及蛋白表达水平显著降低(P均<0.05),细胞活力显著增强(P均<0.05),细胞上清液中IL-6、IL-8、TNF-α表达水平显著降低(P均<0.05)。结论牡荆素可能通过抑制HMGB1/RAGE通路,减轻脂多糖诱导的肺泡上皮细胞炎症损伤。
Objective To investigate the effects of Vitexin on high mobility group box-1 protein/receptor for advanced glycosylation end-products(HMGB1/RAGE)pathway and inflammatory injury induced by lipopolysaccharide(LPS).Methods Human alveolar epithelial cells(HPAEpiC)were cultured in vitro and divided into control group(no treatment),lipopolysaccharide group(10μg/mL lipopolysaccharide),low concentration group(10μg/mL lipopolysaccharide+20μmol/L Vitexin)and high concentration group(10μg/mL lipopolysaccharide+60μmol/L Vitexin).Real-time quantitative PCR(qRT-PCR)was used to detect the mRNA expressions of HMGB1 and RAGE in HPAEpiC cells.Western blot was used to detect the protein expressions of HMGB1 and RAGE in HPAEpiC cells;the proliferation of HPAEpiC cells was detected by MTT method;the levels of interleukin-6(IL-6),interleukin-8(IL-8)and tumor necrosis factor-α(TNF-α)in the supernatant of HPAEpiC cells were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with those in the control group,the expression levels of HMGB1 and RAGE mRNA and protein were significantly higher in LPS group(t=14.269,17.554,19.200,18.887,P<0.05);the cell viability was significantly decreased(t=10.314,P<0.05),and the expression levels of IL-6,IL-8 and TNF-αin cell supernatant were significantly higher(t=45.731,64.827,46.063,P<0.05).Compared with LPS group,with the increase of Vitexin concentration,the expression levels of HMGB1 and RAGE mRNA and protein in HPAEpiC cells were significantly lowered(all P<0.05),the cell viability was significantly increased(all P<0.05),and the expression levels of IL-6,IL-8 and TNF-αin the supernatant were significantly lowered(all P<0.05).Conclusion Vitexin might reduce LPS-induced inflammatory injury of alveolar epithelial cells by inhibiting HMGB1/RAGE pathway.
作者
陈萌
白雨辰
汪铮
CHEN Meng;BAI Yu-chen;WANG Zheng(Department of Respiratory and Critical Care Medicine,Luoyang Central Hospital Affiliated to Zhengzhou University,Luoyang,Henan 471000,China)
出处
《热带医学杂志》
CAS
2021年第2期156-159,165,共5页
Journal of Tropical Medicine
基金
河南省医学科技攻关计划联合共建项目(LHGJ20191214)。
关键词
牡荆素
脂多糖
肺泡上皮细胞
Vitexin
Lipopolysaccharide
Alveolar epithelial cells
