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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 emodin on hypoxia / reoxygenation (H / R) injury in rat hepatocytes and its mechanism. Methods The H / R model of BRL-3A rat hepatocytes was constructed and treated with emodin. Cell viability was detected by CCK-8 assay, apoptosis was detected by flow cytometry, and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were detected by microplate assay. ELISA The levels of TNF-α and IL-1β were detected by real-time quantitative PCR. Toll-like receptor 4 (TLR4), nuclear factor κBp65 (NF- κBp65) mRNA level, Western blot was used to detect TLR4, NF-κBp65 protein levels. Results The survival rate of BRL-3A cells treated with H / R decreased and the apoptosis increased. The levels of ALT, AST, IL-1β and TNF-α in the culture supernatant increased. Reduce the levels of ALT and AST, inhibit the release of IL-1β and TNF-α, and inhibit the increase of H / R-induced TLR4 and NF-κBp65. Conclusion Suedein may reduce H / R injury of rat hepatocytes by regulating TLR4 / NF-κB signaling pathway.