【摘要】 目的研究细胞信号分子糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)对D-氨基半乳糖/脂多糖(D-GalN/LPS)联合诱导的小鼠急性肝衰竭肝细胞凋亡的影响。方法腹腔注射D-GalN/LPS建立小鼠急性肝衰竭模型。实验动物分为对照组、模型组、SB216763干预组(建模前2 h腹腔注射)。检测血清ALT、AST,TUNEL法测定肝细胞凋亡,免疫荧光染色法及比色法检测Caspase-3活性,Western印迹检测Cleaved Caspase-3蛋白表达。多组样本均数的两两比较采用一步法ANOVA分析。结果 GSK-3β活性升高促进了在小鼠急性肝衰竭的发生和发展:抑制GSK-3β活性可以显著改善肝脏功能,血清ALT、AST水平明显下降。SB216763干预组ALT与AST水平分别为(961.1±356.0)IU/L和(2 709.9±423.9)IU/L,SB216763干预组与模型组相比,Caspase-3活性降低,TUNEL方法检测肝细胞凋亡减少,并且Cleaved Caspase-3的蛋白表达降低。结论在D-GalN/LPS诱导的小鼠急性肝衰竭中,抑制GSK-3β活性可能通过抑制肝细胞凋亡而改善肝损伤。因此,对信号分子GSK-3β活性进行干预有可能为急性肝衰竭的治疗提供一个新的靶点。更多还原

【Abstract】 Objective To investigate the effect of glycogen synthase kinase-3β(GSK-3β)on hepatocyte apoptosis in mice with acute liver failure(ALF)induced by injection of D-galactosamine/lipopolysaccharide(D-GalN/LPS).MethodsALF model was established in C57BL/6 mice by intraperitoneal injection of D-GalN/LPS.The mice were divided into control group,ALF model group,and SB216763 treatment group(SB216763in DMSO,i.p,two hours before the induction of ALF).Serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured to assess the liver function.Hepatocyte apoptosis was detected by TUNEL assay,and caspase-3 activity was detected by immunofluorescence staining and colorimetric detection.Western blot was conducted to measure the expression of apoptosisrelated protein caspase 3.One-way ANOVA was carried out in pair-wise comparison of means for multiple samples(homogeneity of variance with LSD-t test,unequal variances with Games-Howell method).Results The increased GSK-3βactivity promoted liver injury in mice with ALF.Compared to ALF model,inhibition of GSK-3βby pretreatment with SB216763 could improve liver function:serum ALT and AST levels decreased significantly.Moreover,when compared to model group,SB216763 treatment group exhibited reduction not only in caspase-3 activity,but also in cleaved caspase-3expression,and even hepatocyte apoptosis.Conclusion In the D-GalN/LPS-induced ALF mice,inhibition of GSK-3βactivity could improve liver injury through reducing hepatocyte apoptosis.Thus,GSK-3βmight be a new target for the treatment of ALF.更多还原