银杏内酯A对LPS诱发肝细胞损伤的保护作用及机制

发布时间：2018-05-03 23:24

本文选题：肝细胞 + 银杏内酯A　；参考：《山东医药》2017年09期

【摘要】：目的探讨银杏内酯A(GA)对脂多糖(LPS)诱发肝细胞损伤的保护作用及可能机制。方法将大鼠肝细胞随机分为6组:正常组、LPS组、LPS+CLI-095组、LPS+GA 20μmol/L组、LPS+GA 10μmol/L组、LPS+GA20μmol/L+LY294002组。MTT法检测细胞活性,Western blotting法检测肝细胞Toll样受体4(TLR4)表达,检测肝细胞NF-κB活性[NF-κB p65、磷酸化-NF-κB p65(p-p65)、磷酸化-IκBα(p-IκBα)、NF-κB p65基因结合活性],检测培养液中炎症细胞因子IL-1β、IL-6、TNF-α水平。结果与正常组比较,LPS组肝细胞TLR4表达增加(P0.05),NF-κB p65、p-p65、p-IκBα水平升高(P均0.05),NF-κB p65基因结合活性增强(P0.05),培养液中IL-1β、IL-6、TNF-α水平升高(P均0.05),细胞活性降低(P0.05)。与LPS组比较,GA剂量依赖性地抑制TLR4表达及NF-κB信号激活(P均0.05),降低上述炎症因子的释放(P均0.05),同时提高细胞活性(P0.05)。TLR4抑制剂CLI-095干预后,LPS引起的肝细胞上述变化被阻断(P0.05)。PI3K/Akt抑制剂LY294002可抵消GA对肝细胞的保护作用(P均0.05)。结论 GA可抑制LPS引起的肝细胞炎症反应、提高细胞活性,其机制可能与抑制LPS引起的TLR-NF-κB信号过度激活有关;PI3K/Akt通路参与了LPS诱发肝细胞损伤过程。
[Abstract]:Objective to investigate the protective effect and possible mechanism of ginkgolide AfGA on lipopolysaccharide (LPS) -induced hepatocyte injury. Methods Rat hepatocytes were randomly divided into six groups: normal group, CLI-095 group, LPS-GA20 渭 mol/L group, LPS-GA10 渭 mol/L LY294002 group, GA20 渭 mol/L LY294002 group. The expression of Toll like receptor 4TLR4 in hepatocytes was detected by Western blotting method. The activity of NF- 魏 B [NF- 魏 B p65, phosphorylated -NF- 魏 B p65p-p65, phosphorylated -I 魏 B 伪 -p-I 魏 B 伪 p65] and the level of IL-1 尾 -IL-6TNF- 伪 in the culture medium were detected. Results compared with the control group, the expression of TLR4 in lipopolysaccharide group increased, and the level of NF- 魏 B p65, p-I 魏 B 伪 increased, the binding activity of NF- 魏 B p65 increased, and the level of IL-1 尾 -IL-6TNF- 伪 increased (P 0.05), and the cell activity decreased (P0.05). Compared with LPS group, GA inhibited TLR4 expression and NF- 魏 B signal activation in a dose-dependent manner (P 0.05), decreased the release of these inflammatory factors (P 0.05), and increased cell activity (P 0.05). TLR4 inhibitor, CLI-095, inhibited these changes in hepatocytes induced by CLI-095. P0.05K / Akt inhibitor LY294002 could counteract the protective effect of GA on hepatocytes. Conclusion GA can inhibit the inflammatory response of hepatocytes induced by LPS and increase the cell activity. The mechanism may be related to the excessive activation of TLR-NF- 魏 B signal induced by LPS and the involvement of PI3K / Akt pathway in the process of hepatocyte injury induced by LPS.
【作者单位】：山东中医药大学附属医院;中国中医科学院西苑医院;
【基金】：山东省中医药管理局基金资助项目(2009-33)
【分类号】：R575

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