漆黄素通过调控TLR4/NF-κB通路减轻大鼠肝细胞缺氧/复氧损伤  

Fisetin alleviates hypoxia/reoxygenation injury in rat hepatocytes via modulation of TLR4/NF-κB signaling pathway

作　　者：蒲俊良 万磊[1] 郑道峰 魏续福[1] 吴忠均[1] 汤成泳[2] 

机构地区：[1]重庆医科大学附属第一医院肝胆外科,重庆400016 [2]重庆医科大学附属第一医院药学部,重庆400016

出　　处：《细胞与分子免疫学杂志》2017年第7期936-941,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81672959);重庆市科委基金(CSTC;2016shms-ztzx0044)

摘　　要：目的探讨漆黄素对大鼠肝细胞缺氧/复氧(H/R)损伤的保护作用及其机制。方法构建BRL-3A大鼠肝细胞H/R模型,并给予漆黄素处理。CCK-8法检测细胞存活率,流式细胞术检测细胞凋亡,微板法检测细胞上清液中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)水平;ELISA检测细胞上清液肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的水平,实时定量PCR检测BRL-3A细胞Toll样受体4(TLR4)、核因子κBp65(NF-κBp65)mRNA水平,Western blot法检测TLR4、NF-κBp65蛋白水平。结果 H/R处理的BRL-3A细胞存活率下降,细胞凋亡增多,培养细胞上清中ALT、AST水平和IL-1β、TNF-α升高;漆黄素处理可提高细胞存活率,减少细胞凋亡,降低ALT、AST水平,抑制IL-1β、TNF-α的释放,同时抑制H/R诱导的TLR4、NF-κBp65水平升高。结论漆黄素可能通过调节TLR4/NF-κB信号通路减轻大鼠肝细胞H/R损伤。Objective To investigate the protective effect of fisetin（ FIS） against hypoxia/reoxygenation（ H/R） injury in rat hepatocytes and its mechanism. Methods H/R injury model of BRL-3A cel s was established and the cel s were pretreated with FIS. Survival rate was detected by CCK-8 assay. Cell apoptosis was measured by flow cytometry. The levels of ALT and AST were determined by microplate assay. The production of TNF-α and IL-1β were detected by ELISA. The mRNA and protein levels of TLR4 and NF-κBp65 were analyzed by quantitative real-time PCR and Western blotting,respectively. Results After subjected to H/R,cell survival rate decreased and the apoptosis level increased. The levels of ALT and AST in cell supernatant were elevated,so were the production of TNF-α and IL-1β. FIS pretreatment increased the cell survival rate and inhibited apoptosis. The levels of ALT,AST and the production of TNF-α and IL-1β were reduced significantly. Moreover,FIS inhibited the increasing expression levels of TLR4 and NF-κBp65 induced by H/R. Conclusion FIS alleviates the hepatocyte injury induced by H/R via modulation of TLR4/NF-κB signaling pathway.

关 键 词：漆黄素 缺氧/复氧 肝细胞损伤 Toll样受体4(TLR4) 核因子κBp65(NF-κBp65) 

分 类 号：R285.5[医药卫生—中药学] R392.33[医药卫生—中医学]