糖原合成酶激酶3活性抑制通过激活PPARα对急性肝衰竭小鼠发挥保护性作用  

Protective effect of glycogen synthase kinase 3β inhibition via peroxisome proliferator-aCTIvated receptor alphaactivation in mice with acute liver failure

作　　者：时红波[1] 时红林[1] 张向颖[1] 陈德喜[1] 段钟平[1] 任锋[1] 

机构地区：[1]首都医科大学附属北京佑安医院北京市肝病研究所,100069

出　　处：《中华肝脏病杂志》2017年第3期211-216,共6页Chinese Journal of Hepatology

基　　金：基金项目：国家自然科学基金（81300349、81270532）;北京市自然科学基金（7172102、7162085、7144216）;北京市科技新星计划（Z131107000413016）;首都临床特色研究（Z161100000516113）;北京市医院管理局临床医学发展专项经费资助（XM201308）;北京市卫生系统高层次卫生技术人才培养项目资助（2014-3-090、2013-3-075）;北京市医院管理局登峰计划专项经费资助（DFL20151601）;国家临床重点专科建设项目（WJWYA-2014-002）

摘　　要：目的用D-氨基半乳糖／脂多糖（D-GaIN／LPS）诱导的小鼠急性肝衰竭模型，探讨糖原合成酶激酶3（GSK3）D和过氧化物酶体增殖物激活受体（PPAR）α信号通路在急性肝衰竭中的作用及其机制。方法以C57BL／6小鼠为研究对象，腹腔注射D-Ga1N／LPS建立小鼠急性肝衰竭模型。用SB216763抑制GSK3p活性，用PPAR仅siRNA抑制PPARα表达。Westernblot检测小鼠中PPARα蛋白表达情况。观察小鼠肝组织病理变化情况以评价肝脏损伤情况，检测血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）评价肝脏功能。实时荧光定量PCR检测肝组织中肿瘤坏死因子（TNF）α、白细胞介素（IL）-1β、IL-12p40mRNA和PPARα基因表达水平。多组样本均数的比较采用单因素方差分析，方差齐者用LSD检验，方差不齐者用Games-Howell法。结果在D-GalN／LPS诱导的肝衰竭小鼠中，抑制GSK3p活性促进了PPARα的mRNA（0．29±0．05与0．17±0．02，F=13．18，P〈0．01）和蛋白（0．79±0．05与0．15±0．04，F=301．36，P〈0．01）表达水平。在D—Ga1N／LPS诱导的急性肝衰竭小鼠中，抑制GSK3β活性减轻了小鼠肝脏出血、炎症和坏死，降低了血清肝功能指标ALT（572．0±127．8与1627．0±412．4，F=25．16，P〈0．01）、AST（479．2±229．2与1359．0±534.8，F=12．96，P〈0．01）水平，也降低了肝组织中炎症因子TNFα（F=32．17，P〈0．01）、IL-1β（F=11．57，P〈0．01）、IL-12β40（F=14．17，P〈0．01）mRNA表达水平。抑制PPARα表达逆转了GSK3p活性抑制带来的肝保护性作用，表现在肝脏出血、炎症和坏死加重，血清肝功能指标AIT（1433．0±464．0与708．7±196．2，F=25．16，P〈0．01）、AST（1361．0±583．2与352．7±177．4，F=12．96，P〈0．01）水平升高，肝组织中炎症因子TNFα（F=32．17，P〈0．01）、IL-1β（F=11．57，P〈0．01）、IL-12p40（F=14．17，P〈0．01）mRNA表Objective To investigate the role of the glycogen synthase kinase 3β （GSK3β） and theperoxisome proliferator-activated receptor alpha （PPARct） signaling pathway in acute liver failure and related mechanisms in a mouse model of acute liver failure induced by D-galactosamine/lipopolysaccharide （D-GaIN/ LPS）. Methods C57BL/6 mice were given intraperitoneal injection of D-GalN/LPS to establish a mouse model of acute liver failure. SB216763 was used to inhibit the activity of GSK3β and PPARα siRNA was used to inhibit the expression of PPARct. Westem blotting was used to measure the expression of PPARα protein. The changes in liver pathology were observed to evaluate liver injury, and the serum levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were measured to assess liver function. Quantitative real-time PCR was used to measure the mRNA expression of tumor necrosis factor-or （TNF-ct）, interleukin-1β （IL-1β）, interleukin-12p40 （IL-12p40）, and PPARα. A one-way analysis of variance was used for comparison of means between multiple groups; the least significant difference test was used for data with homogeneity of variance, and the Games-Howell method was used for data with heterogeneity of variance. Results In the mice with liver failure induced by D-GalN/LPS, GSK3β inhibition promoted the mRNA and protein expression of PPARα （F = 13.18 and 301.36, P = 0.00 and 0.00）. In the mice with acute liver failure induced by D-GalN/LPS, GSK3β inhibition alleviated liver bleeding, inflammation, and necrosis and reduced the serum levels ofALT （F= 25.16,P = 0.000） and AST （F= 12.96,P = 0.001）, as well as the mRNA expression of TNF-α （F= 32.17,P= 0.00）, IL-1β （F= 11.57,P= 0.005）, and IL-12p40 （F= 14.17,P = 0.015） in liver tissue. The inhibition of PPARα expression reversed the liver-protecting effect of GSK3β inhibition, which manifested as aggravation in liver bleeding, inflammation, and necrosis, increases in the serum levels ofALT （F

关 键 词：肝功能衰竭 脂多糖类 炎症 糖原合成酶激酶3 过氧化物酶体增殖物激活受体Α D-氨基半乳糖 

分 类 号：R575.3[医药卫生—消化系统]