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目的 分析D-氨基半乳糖(D-Gal)/脂多糖(LPS)诱导的暴发性肝衰竭模型中肝组织Toll样受体2(TLR2)的表达变化及与细胞因子白细胞介素-18(IL-18)、肿瘤坏死因子α(TNF-α)、γ干扰素(IFN-γ)表达的关系,探讨TLR2在启动炎性应答而致肝损伤中的作用。 方法 BALB/C小鼠腹内联合注射D-Gal 900 mg/kg与LPS 10μg/kg后观察其存活率,并检测不同时间点血清转氨酶和血浆IL-18、TNF-α和IFN-γ含量。用半定量逆转录-聚合酶链反应和Tanon Gis2.0软件分析各时间点肝组织中TLR2 mRNA表达,并与血浆IL-18、TNF-α、IFN-γ含量进行相关分析;免疫组织化学观察肝组织TLR2蛋白的表达。 结果 给药后4 h,血清转氨酶明显升高(与0 h比较,P<0.05);10 h小鼠死亡率达80％。血浆IL-1 8、TNF一α和IFN-γ含量逐步上升,IL-18在1 h即显著升高,之后持续高表达;TNF-α在2 h、5 h有两个分泌高峰;IFN-γ在2 h前增加不明显(F=2.5 7,P=0.1 3),但3 h及以后则显著升高(与0 h比较,P<0.01)。正常小鼠肝组织少量表达TLR2 mRNA,给药后1 h表达即显著增强(与0 h比较,P<0.05);免疫组织化学也显示TLR2蛋白有类似的变化,尤其肝窦内皮细胞、库普弗细胞表达更为显著;且部分肝细胞凋亡、坏死后,残存肝组织仍有较高TLR2表达。相关分析表明,肝组织TLR2 mRNA表达与
Objective To investigate the expression of TLR2 in liver tissue of fulminant hepatic failure induced by D-galactose (L-Gal) / lipopolysaccharide (LPS) and its correlation with the expression of interleukin-18 -18), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ), and to explore the role of TLR2 in initiating inflammatory response to liver injury. Methods BALB / c mice were injected intraperitoneally with D-Gal 900 mg / kg and LPS 10 μg / kg. The survival rate of BALB / c mice was measured. The concentrations of serum transaminase, plasma IL-18, TNF- α and IFN- γ were measured at different time points. The expression of TLR2 mRNA in liver tissue was analyzed by semi-quantitative RT-PCR and Tanon Gis2.0 software, and correlated with the levels of plasma IL-18, TNF-α and IFN-γ. Immunohistochemistry Liver tissue TLR2 protein expression. Results Serum aminotransferase was significantly increased 4 h after administration (compared with 0 h, P <0.05), and the mortality rate reached 80% after 10 h. The content of IL-18, TNF-|Á and IFN-|Ã in plasma gradually increased, IL-18 increased significantly at 1 h and then continued to be high. TNF-|Áproduced two peak at 2 h and 5 h, IFN- γ increased slightly before 2 h (F = 2.57, P = 0.1 3), but increased significantly at 3 h and later (P <0.01 compared with 0 h). The normal mouse liver tissue expressed a small amount of TLR2 mRNA, which was significantly increased 1 h after administration (compared with 0 h, P <0.05). Immunohistochemistry also showed similar changes of TLR2 protein, especially hepatic sinusoidal endothelial cells, The expression of eukaryotic cells was more significant; and some hepatocytes apoptosis, necrosis, the remaining liver tissue is still higher TLR2 expression. Correlation analysis showed that liver tissue TLR2 mRNA expression and