Pu Junliang, Wan Lei, Zheng Daofeng, Wei Xufu, Wu Zhongjun, Tang Chengyong
Department of Hepatobiliary Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China.
Department of Pharmacy, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 Jul;33(7):936-941.
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 cells was established and the cells 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.
目的 探讨非瑟酮(FIS)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 建立BRL-3A细胞H/R损伤模型,并用FIS对细胞进行预处理。采用CCK-8法检测细胞存活率。通过流式细胞术检测细胞凋亡情况。用酶标仪测定ALT和AST水平。采用ELISA法检测TNF-α和IL-1β的产生。分别通过定量实时PCR和蛋白质免疫印迹法分析TLR4和NF-κBp65的mRNA和蛋白水平。结果 H/R处理后,细胞存活率降低,凋亡水平升高。细胞上清液中ALT和AST水平升高,TNF-α和IL-1β的产生也增加。FIS预处理提高了细胞存活率并抑制了凋亡。ALT、AST水平以及TNF-α和IL-1β的产生均显著降低。此外,FIS抑制了H/R诱导的TLR4和NF-κBp65表达水平的升高。结论 FIS通过调节TLR4/NF-κB信号通路减轻H/R诱导的肝细胞损伤。