过氧化物酶体增殖物激活受体α对急性肝衰竭小鼠内质网应激的影响  

Effect of peroxisome proliferator activated receptor α on endoplasmic reticulum stress in acute liver failure mice

作　　者：宋金钥 张向颖[2] 时红波[2] 陈德喜[2] 段钟平[2] 张桓虎[1] 任锋[2] 

机构地区：[1]山西医科大学第二医院消化科,太原030001 [2]首都医科大学附属北京佑安医院肝病研究所,北京100069

出　　处：《中华危重症医学杂志（电子版）》2017年第1期3-8,共6页Chinese Journal of Critical Care Medicine:Electronic Edition

基　　金：国家自然科学基金项目(81270532);北京市自然科学基金项目(7162085);首都特色临床应用研究项目(Z121107001012167);北京市卫生系统高层次卫生技术人才培养计划项目(2013-3-075)

摘　　要：目的研究过氧化物酶体增殖物激活受体α(PPARα)通过调控内质网应激(ERS)在小鼠急性肝衰竭(ALF)肝损伤病理机制中的作用。方法腹腔注射D-氨基半乳糖(D-Gal N)和脂多糖(LPS)诱导小鼠ALF模型。将小鼠分为对照组(仅给予相应量的磷酸盐缓冲液,10只)、模型组(16只)和Wy-14643干预组(造模前2 h以6 mg/kg通过尾静脉注射,16只)。同时,将模型组进一步分为给药后1、3、6 h 3个亚组,比较各亚组与对照组间内质网应激特异性凋亡蛋白CCAAT/增强子结合蛋白同源蛋白(CHOP)和PPARα表达情况。建模6 h后,检测血清丙氨酸转氨酶(ALT)、天门冬酸氨基转移酶(AST)评价肝脏功能。采用免疫印迹技术比较对照组、模型组及Wy-14643干预组间caspase-3、cleaved caspase-3、CHOP表达情况。另取13只小鼠为4-丁酸苯酯(4-PBA)干预组(建模前6 h腹腔注射给予100 mg/kg的4-PBA),比较对照组、模型组及4-PBA干预组间PPARα蛋白及m RNA表达情况。结果与对照组相比,随着ALF的逐渐进展,模型组给药后3、6h CHOP表达显著升高(F=6.341,P=0.025),PPARα表达显著降低(F=7.115,P=0.022)。三组小鼠间血清ALT、AST、caspase-3、cleaved caspase-3和CHOP表达水平比较,差异均有统计学意义(F=8.454,P=0.027;F=10.252,P=0.016;F=6.231,P=0.042;F=30.072,P<0.001;F=8.596,P=0.014)。Wy-14643干预组血清ALT[(524±330)U/L vs.(1 465±485)U/L]、AST[(1 227±314)U/L vs.(4 038±1 537)U/L]水平均显著低于模型组(P均<0.05),且与模型组相比,对照组与Wy-14643干预组小鼠caspase-3显著升高,cleaved caspase-3和CHOP表达显著降低(P均<0.05)。与此同时,4-PBA干预组PPARα的m RNA和蛋白表达均显著高于模型组(F=6.665,P=0.017;F=5.441,P=0.043)。结论 PPARα可能通过抑制严重内质网应激而保护小鼠急性肝衰竭后肝损伤。Objective To study the role of peroxisome proliferator activated receptor α（PPARα） on serious endoplasmic reticulum stress in acute liver failure（ALF） mice induced by D-Galactosamine/lipopolysaccharide（D-Gal N/LPS）. Methods ALF model was established by intraperitoneal injection of D-Gal N/LPS in C57BL/6 mice. Animal experimental groups included the control group（corresponding volume phosphate buffered saline, 10 mice）, model group（16mice）, Wy-14643 group（6 mg/kg Wy-14643 by tail vein on 2 h before model establishment）.Meanwhile, mice in the model group further divided into three subgroup according to 1, 3, 6 h after D-Ga N/LPS injection. The expression of C/EBP homologous protein（CHOP） and peroxisome proliferator activated receptor α（PPARα） among subgroup and the control group were compared.The levels of alanine transaminase（ALT） and aspartate aminotransferase（AST） were detected, the expression of caspase-3, cleaved caspase-3 and CHOP were examined by Western-blotting on 6 h after model establishment. The other 13 mice were injected 100 mg/kg 4-phenylbutyrate（4-PBA） on 6 h before model establishment as the 4-PBA group. The PPARα protein and m RNA were detected and compared. Results In the model group on 3, 6 h after D-Ga N/LPS injection,the expression of CHOP increased（F = 6.341, P = 0.025）, and PPARα decreased（F = 7.115, P = 0.022）as compared with those in the control group during the progression of ALF. The ALT, AST,caspase-3, cleaved caspase-3 and CHOP among the control group, model group and Wy-14643 group all showed significant differences（F = 8.454, P = 0.027; F = 10.252, P = 0.016; F = 6.231, P= 0.042; F = 30.072, P〈0.001; F = 8.596, P = 0.014）. And the levels of ALT [（524 ± 330） U/L vs.（1 465 ± 485） U/L] and AST [（1 227 ± 314） U/L vs.（4 038 ± 1 537） U/L] in the Wy-14643 group were much lower than those in the model group（all P〈0.05）. In the control group and Wy-14643 group, the expression of caspase-3 increased marke

关 键 词：过氧化物酶体增殖物激活受体Α 肝功能衰竭 急性 内质网 小鼠 

分 类 号：R575.3[医药卫生—消化系统]