CAJ | 학술논문

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（2709．9±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。
목적：연구세포신호분자당원합성매격매-3β（glycogen synthase kinase-3β，GSK-3β）대 D-안기반유당／지다당（D-GalN／LPS）연합유도적소서급성간쇠갈간세포조망적영향。방법복강주사 D-GalN／LPS 건립소서급성간쇠갈모형。실험동물분위대조조、모형조、SB216763간예조（건모전2 h 복강주사）。검측혈청 ALT、AST，TUNEL 법측정간세포조망，면역형광염색법급비색법검측 Caspase-3활성，Western 인적검측 Cleaved Caspase-3단백표체。다조양본균수적량량비교채용일보법 ANOVA 분석。결과 GSK-3β활성승고촉진료재소서급성간쇠갈적발생화발전：억제 GSK-3β활성가이현저개선간장공능，혈청 ALT、AST 수평명현하강。SB216763간예조 ALT 여 AST 수평분별위（961．1±356．0）IU／L화（2709．9±423．9）IU／L，SB216763간예조여모형조상비，Caspase-3활성강저，TUNEL 방법검측간세포조망감소，병차Cleaved Caspase-3적단백표체강저。결론재 D-GalN／LPS 유도적소서급성간쇠갈중，억제 GSK-3β활성가능통과억제간세포조망이개선간손상。인차，대신호분자 GSK-3β활성진행간예유가능위급성간쇠갈적치료제공일개신적파점。
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).Methods ALF 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 (SB216763 in 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 apoptosis-related 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-3β by 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-3 expression,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-3β might be a new target for the treatment of ALF.