酵母抽提物改善酒精和脂多糖诱导的HepG2细胞炎症反应
Yeast extract improves the inflammatory response of HepG2 cells induced by ethyl alcohol and lipopolysaccharide
田蕾 1张彦 2程倩 3韩艳阳 1董亚静 1李想 1韩浩1
作者信息
- 1. 山西医科大学公共卫生学院营养与食品卫生学教研室,太原 030001
- 2. 酵母功能湖北省重点实验室,宜昌 443000
- 3. 酵母功能湖北省重点实验室,宜昌 443000;宜昌市营养健康食品工程技术研究中心,宜昌 443000
- 折叠
摘要
目的 探讨酵母抽提物对酒精与脂多糖联合诱导的人肝癌细胞HepG2炎症反应的改善作用,并基于Toll样受体4(Toll-like receptor 4,TLR4)/核因子kappa B(nuclear factor kappa-B,NF-κB)信号通路进一步探索潜在分子机制.方法 采用50 mmol/L酒精+1 μg/mL脂多糖联合诱导HepG2细胞发生炎症反应,同时给予0.5 mmol/L酵母抽提物干预.ELISA法检测细胞炎症因子表达水平;免疫荧光染色检测TLR4表达水平及NF-κB核转位情况;蛋白免疫印迹检测TLR4、NF-κB、磷酸化NF-KB-p65(phospho-nuclear factor kappa-B-p65,P-NF-KB-p65)、核内磷酸化 NF-κB-p65(nucleus-phospho-nuclear factor kappa-B-p65,N-P-NF-KB-p65)、肿瘤 坏死因 子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)和白细胞介素-1β(interleukin-1β,IL-1 β)的表达水平.结果 与对照组相比,模型组细胞培养上清液中 1L-6、IL-1β 和 TNF-α 水平显著增加(P＜0.05),TLR4、细胞核 NF-κB、P-p65 及 N-P-p65蛋白表达均显著上调(P＜0.05).与模型组相比,酵母抽提物干预组IL-6、IL-1β和TNF-α水平显著降低(P＜0.05),肝细胞TLR4表达水平下调(P＜0.05),并抑制NF-κB核转位(P＜0.05).结论 酵母抽提物能有效抑制酒精+脂多糖联合诱导的HepG2细胞炎症反应,其分子机制可能与下调TLR4/NF-κB通路有关.
Abstract
OBJECTIVE To explore the ameliorative effect of yeast extract(YE)on the inflammatory response of human hepatoma cells(HepG2)induced by ethyl alcohol(EtOH)and lipopolysaccharide(LPS),and further explore the potential molecular mechanism based on Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)signaling pathway.METHODS HepG2 cells were induced by 50 mmol/L EtOH and 1 μg/mL LPS combined with YE intervention.The expression level of inflammatory cytokines was detected by ELISA.The expression level of TLR4 and the nuclear translocation of NF-κB were detected by immunofluorescence staining.The expression levels of TLR4,NF-κB,phospho-NF-KB-P65(P-NF-KB-p65),nucleus-phospho-NF-KB-p65(N-P-NF-KB-p65),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1 β(IL-1β)were detected by Western blot.RESULTS Compared with the control group,the cells in EtOH+LPS group produced a large number of inflammatory factors and had a significant inflammatory response.YE intervention significantly alleviated EtOH+LPS induced hepatocyte inflammatory response.Further molecular mechanism studies showed that YE significantly reduced TLR4 expression level and inhibited NF-κB nuclear translocation in hepatocytes.CONCLUSION YE can effectively inhibit the inflammatory response of HepG2 cells induced by EtOH and LPS,and its molecular mechanism may be related to the down-regulation of TLR4/NF-κB pathway.
关键词
酵母抽提物/酒精/脂多糖/Toll样受体4/核因子kappa/BKey words
yeast extract/ethyl alcohol/lipopolysaccharide/toll-like receptor 4/nuclear factor kappa B引用本文复制引用
出版年
2024
NETL
NSTL
维普
万方数据