Objective To investigate the effect and mechanism of thymoquinone on hepatic ischemia-reperfusion injury (IRI).Methods Thirty C57 mice were randomly divided into the sham operation (sham),IRI and thymoquinone (Thy)groups (n =1 0 in each group).At preoperative 1 h,thymoquinone at a dose of 40 ml/kg was administered via intraperitoneal injection in the Thy group.Absolute ethyl alcohol at the same dosage was given via intraperitoneal injection in the sham and IRI groups.Liver IRI mouse models were established in the IRI and Thy groups.Serum and liver specimens were collected at 4 h after reperfusion.Under light microscope,hepatic histopathological changes were observed and assessed by pathological injury grading.Reverse transcription polymerase chain reaction (RT-PCR)was performed to measure the messenger ribonucleic acid (mRNA)expression levels of tumor necrosis factor (TNF)-α,monocyte chemotactic protein (MCP)-1 and interleukin (IL)-6.The expression of TNF-α,MCP-1 and IL-6 proteins in the serum in the serum were assessed by ELISA.The content of malondialdehyde (MDA)in the liver tissue was detected by thiobarbituric acid (TBA).The activity of catalase (CAT),glutathioneperoxidase (GPx)and superoxide dismutase (SOD)in the liver tissue was determined by ELISA.The expression levels of Wnt,β-catenin and p53 proteins were measured by Western blot.Results Compared with the sham group,the liver injury was more severe and the hepatic injury grading was significantly enhanced in the IRI group (P <0.05),the expression of TNF-α,MCP-1 and IL-6 in the liver tissue and serum sample,and MDA, Wnt,β-catenin and p53 in the liver tissue was significantly up-regulated (P <0.05-0.001 ),whereas CAT,GPx and SOD activity in the liver tissue was dramatically reduced (all in P <0.001 ).Compared with the IRI group,the liver injury in the Thy group was slighter and the liver injury grading was significantly decreased (both in P <0.05).The expression levels of TNF-α,MCP-1 and IL-6 in the liver tissue and serum sample,MDA,Wnt,β-catenin and p53 in the liver tissue were significantly down-regulated (all in P <0.05),whereas CAT,GPx and SOD activity was considerably up-regulated in the liver tissue (all in P <0.05).Conclusions Thymoquinone can mitigate liver IRI through alleviating inflammatory response and oxidation stress.The underlying mechanism is correlated with inhibiting the activation of Wnt/β-catenin/p53 signaling pathway.%目的：探讨百里醌对肝缺血-再灌注损伤（IRI）的作用及其机制。方法30只 C57小鼠随机均分为假手术（Sham）组、IRI 组和百里醌（Thy）组（每组各10只）。术前1 h，Thy 组给予百里醌（40 ml／kg）腹腔注射，Sham 组和 IRI 组给予无水乙醇（40 ml／kg）腹腔注射。IRI 组和 Thy 组建立小鼠肝 IRI 模型。再灌注4 h后收集血清和肝脏标本。光学显微镜下观察肝组织病理学改变，并予病理损伤评分；采用逆转录聚合酶链反应（RT-PCR）检测肝组织肿瘤坏死因子（TNF）-α、单核细胞趋化蛋白（MCP）-1和白细胞介素（IL）-6的信使核糖核酸（mRNA）表达水平；采用酶链免疫吸附试验（ELISA）检测血清中 TNF-α、MCP-1和 IL-6的蛋白表达水平；采用硫代巴比妥酸（TBA）法检测肝组织丙二醛（MDA）的含量；采用 ELISA 法测定肝组织过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GPx）、超氧化物歧化酶（SOD）活性；采用肝组织免疫印迹法（Western blot）检测 Wnt、β-catenin、p53的蛋白表达水平。结果与 Sham 组比较，IRI 组肝组织损伤严重，损伤评分明显增加（P ＜0.05），肝组织和血清中的 TNF-α、MCP-1、IL-6和肝组织 MDA、Wnt、β-catenin、p53的表达均明显增多（P ＜0.05～0.001），而肝组织 CAT、GPx 与 SOD 活性均明显降低（均为 P ＜0.001）。与 IRI 组比较， Thy 组肝组织损伤较轻，损伤评分明显减少（P ＜0.05），肝组织和血清中的 TNF-α、MCP-1、IL-6和肝组织MDA、Wnt、β-catenin、p53的表达均明显减少（均为 P ＜0.05），而肝组织 CAT、GPx 与 SOD 活性均明显增高（均为 P ＜0.05）。结论百里醌通过减轻炎症反应和氧化应激而减轻肝 IRI，其作用机制与抑制 Wnt／β-catenin／p53信号通路激活有关。
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