摘要
目的 研究过氧化物酶体增殖物激活受体α（PPAR α）对D-氨基半乳糖（D-GalN）联合脂多糖（LPS）诱导的小鼠急性肝衰竭（ALF）的保护作用及其机制.方法 第一部分实验将C57BL/6小鼠按随机数字表法随机分为对照组、ALF 2h组、ALF 4h组、ALF 6h组,每组8只;第二部分实验将C57BL/6小鼠按随机数字表法随机分为对照组、ALF组、WY14643组,每组8只.前后两部分实验的给药方式一样,通过腹腔注射溶于生理盐水中的D-GalN （700 mg/kg）联合LPS（10 μg/kg）构建小鼠ALF模型;WY14643作为PPARα的选择性激动剂于造模前2h以6 mg/kg通过尾静脉注射.第一部分实验分别在D-GalN/LPS注射后及注射后2h、4h、6h麻醉小鼠取血,获取肝组织并迅速提取mRNA;第二部分实验在D-GalN/LPS注射后6h麻醉小鼠取血,获取肝组织并迅速提取mRNA,mRNA和肝组织都在-80℃储存供以后分析使用.组织病理学分析及血清转氨酶活性测定观察肝组织损伤的严重程度;实时定量PCR检测促炎性细胞因子TNFα、IL-1β、IL-6以及趋化因子（CXCL）-1、CXCL-10的mRNA表达;蛋白质印迹检测炎症信号通路相关蛋白（磷酸化的NF-κBp65、JNK、ERK、p38）的表达.结果 PPAR α的mRNA和蛋白在急性肝衰竭疾病进展过程中表达逐渐减少.在给予WY14643激活PPAR α后,与ALF组相比,WY14643组肝小叶结构较完整,部分肝细胞呈气球样变,小叶内及汇管区炎细胞浸润不明显,血清ALT、AST水平较低[ALT：WY14643组（555±62）U/L,ALF组（2 898±822） U/L,P＜0.05;AST：WY14643组（791±58） U/L,ALF组（3 013±997）U/L,P＜0.05],促炎性细胞因子、趋化因子的mRNA及磷酸化的NF-κB p65、ERK、JNK蛋白表达减少,且差异具有统计学意义.在对照组、ALF组、WY14643组基因表达结果显示,TNFα的相对表达量分别为0.161±0.085、7.996±1.068、3.346±0.942,P＜0.05;IL-1β的相对表达量分别为0.041±0.002、3.657±0.904、0.176±0.089,P＜0.01;IL-6的相对表�
Objective To determine the role and mechanism of peroxisome proliferator activated receptors （PPAR） α in a mouse model of D-galactosamine/lipopolysaccharide （D-GalN/LPS）-induced acute liver failure（ALF）.Methods Firstly,C57BL/6 mice were randomly divided into control group（n =8）,ALF 2h group（n =8）,ALF 4h group （n =8）,ALF 6h group （n =8）.Secondly C57BL/6 mice were randomly divided into control group（n =8）,ALF group（n =8）,WY14643 group（n =8）.To induce ALF,the mice were injected intraperitoneally with D-GalN （700 mg/kg） and LPS （10 μg/kg）.WY14643 （6 mg/kg）,the selective agonist of PPAR α,was administered via tail vein two hours prior to D-GalN/LPS exposure.Two,four,and six hours after D-GalN/LPS treatment in the first study,mice were anesthetized and blood was collected,6h after D-GalN/LPS treatment in the second study,blood was collected.The liver tissue was harvested for histology and mRNA extraction.Serum levels of ALT and AST were measured to evaluate the hepatic damage.Inflammatory cytokines （TNFα,IL-1β,IL-6） and chemokines （CXCL-1,CXCL-10） were detected by real-time quantitative PCR.Differential protein expression of p-NF-κBp65,p-JNK,p-ERK,p-p38 in inflammatory pathways was detected by Western blotting.Significance of inter-group differences was assessed by one-way ANOVA,and pairwise comparison was performed by the least significant difference test.Results The gene and protein expression of PPAR α were gradually reduced during the development of ALF.Compared with the model group,the liver architecture was better preserved almost with normal morphology in WY14643-treated mice.Serum ALT and AST levels in WY14643-treated group were significantly lower [ALT：（555 ±62）U/L vs （2 898 ±822） U/L,P 〈0.05; AST：（791 ±58） U/L vs （3 013 ±997）U/L,P 〈 0.05].The expression of proinflammatory cytokines and chemokines was significantly suppressed during the activation of PPAR α.In the second study,the levels of gene expression of proinf
出处
《中华内科杂志》
CAS
CSCD
北大核心
2014年第9期730-734,共5页
Chinese Journal of Internal Medicine
基金
国家“十二五”科技重大专项课题（2012ZX10002004-006）
