摘要
该研究探讨人参皂苷Rg1对非酒精性脂肪性肝细胞炎症反应的作用及其分子机制。用1 mmol/L游离脂肪酸处理HepG2和L02细胞24 h,再用20μg/mL或40μg/mL人参皂苷Rg1处理6 h;设置对照组、模型组、低剂量Rg1组、高剂量Rg1组。全自动生化仪检测各组细胞上清谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)的含量;酶联免疫吸附法测定细胞上清IL-1β、IL-6、TNF-α。RT-qPCR及Western blot检测NF-κB通路相关基因及蛋白的改变。免疫荧光染色观察NF-κB核转移;Western blot检测各组胞质与胞核内的NF-κB P65蛋白的表达。与对照组相比,模型组培养上清炎症指标明显增加(P<0.05);Rg1能降低炎症指标的表达(P<0.05)。Rg1能减少游离脂肪酸诱导的NF-κB磷酸化及其下游IL-1β、IL-6、TNF-α的表达,减少NF-κB P65从胞质向胞核的转移(P<0.05)。Rg1可通过抑制NF-κB活化减少NASH细胞模型炎症反应,为非酒精性脂肪性肝炎的治疗提供了可能的靶点。
This work was to investigate the effect of ginsenoside Rg1 on inflammation in NASH(non-alcoholic steatohepatitis)cell model and its molecular mechanism.HepG2 cells and L02 cells were treated with 1 mmol/L FFA(free fatty acid)for 24 h,and then treated with 20μg/mL or 40μg/mL ginsenoside Rg1 for 6 h.The control group,model group,low-dose Rg1 group and high-dose Rg1 group were set.The ALT(alanine aminotransferase)and AST(aspartate aminotransferase)in supernatant were detected by automatic biochemical analyzer.IL-1β,IL-6,TNF-αin supernatant were detected with ELISA(enzyme-linked immunosorbent assay).RT-qPCR and Western blot were used to detect alterations of genes and proteins related to NF-κB pathway.Immunofluorescence was used to demonstrate NF-κB P65 nuclear translocation.and Western blot was used to detect the expression of NF-κB P65 protein in the cytoplasm and nucleus of each group.Compared with the control group,the inflammatory cytokines in supernatant of the model group were significantly increased(P<0.05).Rg1 could decrease the expressions of inflammatory indicators(P<0.05).Rg1 could down-regulate FFA activated NF-κB phosphorylation,translocation of NF-κB P65 from cytoplasm to nucleus,and the downstream target genes of NF-κB,including IL-1β,IL-6 and TNF-α(P<0.05).Rg1 might alleviate FFA mediated inflammation in NASH cell model through inhibiting NF-κB activation,which provided a possible target for NASH treatment.
作者
肖晴
章述军
阳成
高月
徐静
朱雅莉
黄文祥
XIAO Qing;ZHANG Shujun;YANG Cheng;GAO Yue;XU Jing;ZHU Yali;HUANG Wenxiang(Department of Infectious Diseases,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2020年第1期102-109,共8页
Chinese Journal of Cell Biology
基金
重庆市渝中区科学技术局基础研究与前沿探索项目(批准号:20190138)资助的课题。
