摘要
目的探讨细胞内信号分子糖原合成酶激酶(GSK)30在HBV引起的重型肝炎肝衰竭发生过程中的作用。方法收集2009年至2011年北京佑安医院就诊患者中慢性乙型肝炎患者(12例,慢乙型肝炎患者组)和HBV感染引起的重型肝炎肝衰竭患者(12例,乙型重型肝炎肝衰竭组)以及正常人(8例,对照组)的肝组织资料,进行各项临床检测指标的统计分析,用细胞裂解液匀浆提取蛋白,用GSK3D活性测定试剂盒检测肝组织中GSK3D活性,用Westernblot方法检测p-GSK3、GSK3、β-肌动蛋白的表达;制备石蜡切片进行免疫荧光检测。用LSD-t检验进行组间比较,P〈0.05为差异有统计学意义。结果Westernblot结果显示,与对照组相比,慢性乙型肝炎患者组GSK3β第9位丝氨酸磷酸化程度上升,活性下降,重型肝炎肝衰竭患者组GSK3β第9位丝氨酸磷酸化程度显著下降,即GSK3β活性显著上升(P=0.0342);同时,免疫荧光方法对各组样本进行检测结果显示乙型重型肝炎肝衰竭组GSK3D磷酸化水平显著下降,证明其活性明显增高;最后,GSK3D活性检测试剂盒进行GSK3β活性检测,与Westernblot和免疫荧光结果相一致,重型肝炎肝衰竭患者中GSK3β涪性显著增加,与对照组相比,P=0.0289。结论GSK3活性在重型肝炎肝衰竭发生过程中被激活,因此其是重型肝炎肝衰竭发病机制中的一种重要信号分子,抑制其活性可能在重型肝炎肝衰竭的预防和治疗中发挥一定的作用。
Objective To investigate the role of glycogen synthase kinase-3β (GSK3β) in the development of severe hepatitis liver failure (SHLF) caused by the hepatitis B virus. Methods Twelve patients with chronic hepatitis B (CHB) (CHB group), 12 patients with SHLF caused by hepatitis B virus (SHLF group), and 8 normal subjects (control group), who were admitted to Beijing You'an Hospital from January 2009 to December 2011, were included in this study. Their liver tissues were collected to do some clinical examinations. The GSK3β activity in the liver tissue was detected with a GSK3β activity assay kit. Western blot was used to determine the expression of p-GSK3, total GSK3, and -actin. The paraffin sections of livertissue were prepared for immunofluorescence assay. All data were expressed as mean:kstandard deviation, and comparison between groups was made by least significant difference t test. P 〈 0.05 was considered statistically significant. Results Western blot results showed that compared with the control group, the CHB group had a higher level of p-GSK313 and the SHLF group had a significantly lower level of p-GSK3β (P = 0.0342). The immunofluorescence assay results showed that the SHLF group had a significantly lower level of p-GSK3β than the control group. GSK3β activity assay results showed that compared with the control group, the CHB group had a significantly lower GSK3β activity and the SHLF group had a significantly higher GSK3β activity (P = 0.0289), which were consistent with the results of Western blot and immunofluorescence assay. Conclusion GSK3 is activated in the development of SHLF, so it is an important signaling molecule in the pathogenesis of SHLF. Inhibiting its activity may play a role in the prevention and treatment of SHLF.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2016年第4期265-269,共5页
Chinese Journal of Hepatology
基金
基金项目:国家十二五科技重大专项(2012ZX10002004-006、2012ZX10004904-003-001,2013ZX10002002-006-001)
国家自然科学基金项目(81270532)
北京市优秀人才培养资助(D类,2011D003034000022)
2012北京市留学人员科技活动择优资助项目
首都临床特色应用研究(Z121107001012167)
王宝恩肝纤维化研究基金项目(CFHPC20131031)