摘要
目的探讨甘草酸二铵对小鼠急性肝衰竭发生过程中小鼠肝组织高迁移率族蛋白B1(HMGB1)表达的影响。方法取雄性昆明种小鼠72只,随机分为正常对照组、急性肝功能衰竭组及甘草酸二铵治疗组。模型组及治疗组予D-Galn 500mg/kg及LPS 10μg/kg腹腔注射建立急性肝功能衰竭小鼠模型,并于建模1h后予以甘草酸二铵70mg/kg对治疗组小鼠进行灌胃处理,正常对照组及模型组予等量生理盐水灌胃。以建模后4,8,12,24h分别取静脉血以常规生化方法检测小鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)水平,HE染色法检查肝组织病理学变化,采用免疫组织化学法及实时荧光定量PCR检测小鼠肝组织中HMGB1的蛋白表达量及mRNA水平。结果模型组及治疗组小鼠血清ALT和AST水平均显著高于正常对照组(P<0.01),且模型4,8h组小鼠血清ALT和AST含量均高于相应治疗组(P<0.05),模型8,12,24h组及甘草酸二铵治疗12,24h组小鼠血清TBIL水平均显著高于正常对照(P<0.05)。免疫组化结果发现,模型8,12,24h组及治疗8,12,24h组小鼠肝组织中HMGB1蛋白表达水平较相应正常对照组显著增加(P<0.01),且随肝衰竭发展明显增加,模型8,12h组HMGB1蛋白表达量高于相应甘草酸二铵治疗组(P<0.05)。Real time-PCR结果显示,模型组和治疗8,12,24h组小鼠肝组织HMGB1mRNA表达量高于正常对照组(P<0.05),模型8h、12h组HMGB1mRNA表达量高于相应治疗组(P<0.05)。结论 HMGB1在小鼠急性肝功能衰竭过程中发挥了重要作用。甘草酸二铵可能通过抑制HMGB1的产生减轻肝损伤。
Objective To observe the expression influence of transcription factor HBGB1 in the process of acute liver failure induced by D-GalnandLPS,and explore the possible role of transcription factor HMGB1 in the pathogenesis of mice acute hepatic failure.Methods Total of 72 male mice was randomly divided into normal control group,acute liver failure model group,and glycyrmizinate acid diamine therapy group.Each group except the normal group was injected with D-GalN 500mg/kg and LPS 10μg/kg intraperitoneally,and after one hour,the diammonium glycyrmizinate group was fed with the glycyrrhizic acid diamine 70mg/kg and other two groups were fed with normal saline(NS).Liver tissue was taken and detected by using HE staining method to check liver tissue pathological changes after 4h,8h,12 h,24h.The expression of HMGB1 protein in hepatic tissue was detected by immunohistochemistry and the expression of HMGB1 mRNA was determined by real time-PCR.Results Immunohistochemistry results was shown that model group and therapy group mice liver tissue HMGB1 protein expression at 8h,12 h,and 24 h were higher than normal control group,and there were statistical significance differences(P〈0.01),and the model group of HMGB1 protein expression at 8hand 12 hwas higher than therapy group(P〈0.05).Real time PCR results was shown that model and therapy groups' s HMGB1 mRNA expression at 8h,12 h,and 24 hwere higher than normal control group(P〈0.05),and the model group's HMGB1 mRNA expression at 8h,12 h was higher than the therapy group(P〈0.05).Conclusion HMGB1 in the mice with acute liver failure model may play in a key role.Glycyrrhizic acid diamine can inhibit HMGB1 of the liver cell to protect liver.
出处
《贵州医药》
CAS
2016年第5期456-459,共4页
Guizhou Medical Journal