前列地尔对暴发性肝衰竭大鼠eIF2α/ATF4/CHOP通路及肝功能的影响  

Effects of alprostadil on the eIF2α/ATF4/CHOP pathway and liver function in rats with fulminant hepatic failure

作　　者：李盼盼 符健[1] 陈伟[2] 陈丽 LI Panpan;FU Jian;CHEN Wei;CHEN Li(Department of Infection,Hainan Provincial People’s Hospital(Hainan Hospital Affiliated to Hainan Medical College),Haikou 570100,China;Department of Pharmacy,the Second Affiliated Hospital of Hainan Medical College,Haikou 570100)

机构地区：[1]海南省人民医院(海南医学院附属海南医院)感染科,海口570100 [2]海南医学院第二附属医院药学部,海口570100

出　　处：《中国比较医学杂志》2021年第8期55-62,共8页Chinese Journal of Comparative Medicine

基　　金：海南省卫生计生行业科研项目(18A200189)。

摘　　要：目的探究前列地尔(PGE1)对暴发性肝衰竭(FHF)大鼠对肝功能的影响,并探讨其对真核翻译起始因子2α(eIF2α)/激活转录因子4(ATF4)/C/EBP同源蛋白(CHOP)通路的调控作用。方法SPF级SD雄性大鼠90只按随机数表法分为对照组、模型组、阳性对照组、低、中、高剂量PGE1组,除对照组外,其它组大鼠均采用腹腔注射D-氨基半乳糖(D-GalN)-大肠杆菌内毒素脂多糖(LPS)的方法建立FHF大鼠模型,对照组腹腔注射等量的生理盐水。造模6 h后,阳性对照组及低、中、高剂量PGE1组分别尾静脉注射促肝细胞生长素1.36 mg/kg,PGE112.5、25、37.5μg/kg,每天1次,连续给药3 d,对照组和模型组尾静脉注射等量的生理盐水。在造模72 h后处死大鼠,腹主动脉采血,检测血清丙氨酸氨基转氨酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)水平;解剖取肝组织用苏木精-伊红(HE)染色法观察各组大鼠肝组织病理学变化;采用实时荧光定量PCR(qRT-PCR)和蛋白免疫印迹(Western blot)法检测肝组织中eIF2α/ATF4/CHOP/半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)mRNA和蛋白、磷酸化-eIF2α(p-eIF2α)蛋白水平。结果与对照组相比,模型组肝细胞广泛变性并有局灶性坏死,中央静脉受损,血清ALT、AST、TBIL,肝组织中eIF2α、ATF4、CHOP、Caspase-3 mRNA及p-eIF2α/eIF2α、ATF4、CHOP、Caspase-3蛋白水平均升高(P<0.05);与模型组相比,阳性对照组、低、中、高剂量PGE1组肝细胞损害有所降低,坏死细胞数量减少,血清ALT、AST、TBIL水平,肝组织中eIF2α、ATF4、CHOP、Caspase-3 mRNA及p-eIF2α/eIF2α、ATF4、CHOP、Caspase-3蛋白水平降低(P<0.05);且随着PGE1给药剂量的增加,低、中、高剂量PGE1组血清ALT、AST、TBIL水平,肝组织中eIF2α、ATF4、CHOP、Caspase-3 mRNA及p-eIF2α/eIF2α、ATF4、CHOP、Caspase-3蛋白水平依次降低(P<0.05),呈剂量依赖性;与阳性对照组相比,低、中剂量PGE1组血清ALT、AST、TBIL水平,肝组织中eIF2�Objective To investigate the effects of prostaglandin E1(PGE1)on liver function in rats with fulminant hepatic failure(FHF),and to explore its regulatory effects on the eukaryotic translation-initiation factor 2α(eIF2α)/activating transcription factor 4(ATF4)/C/EBP homologous protein(CHOP)pathway.Methods Overall,90 specific pathogen-free Sprague-Dawley rats were divided into control,model,positive control,low-,medium-,and highdose PGE1 groups according to the random number table method.Except for the control group,all rats were intraperitoneally injected with D-galactosamine(D-GalN)-lipopolysaccharide(LPS)to establish the FHF rat model,and the control group received an intraperitoneal injection of the same volume of normal saline.At 6 hours after modeling,the positive control group and low-,medium-,and high-dose PGE1 groups were administered a tail vein injection of 1.36 mg/kg hepatocyte growth promoting factor and 12.5,25,or 37.5μg/kg PGE1,respectively,once a day for 3 consecutive days.The control group and the model group were administered a tail vein injection of the same volume of normal saline.The rats were euthanized at 72 hours after modeling and abdominal aorta blood was collected.The serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(TBIL)were measured.Liver tissues were dissected and stained with hematoxylin and eosin(HE)to observe pathological changes.mRNA and protein levels of eIF2α/ATF4/CHOP/caspase-3 were detected by real-time fluorescence quantitative PCR(qRT-PCR)and Western blot was used to measure phosphorylated-eIF2α(p-eIF2α)protein levels.Results Compared with the control group,hepatocytes in the model group showed extensive degeneration and focal necrosis,and the central venous was damaged.The levels of ALT,AST,and TBIL in serum,mRNA levels of eIF2α,ATF4,CHOP,and caspase-3,and protein levels of p-eIF2α/eIF2α,ATF4,CHOP,and caspase-3 in liver tissues were higher(P<0.05).Compared with the model group,the damage to hepatocytes and the number

关 键 词：前列地尔 暴发性肝功能衰竭 大鼠 真核翻译起始因子2α 激活转录因子4 C/EBP同源蛋白 

分 类 号：R-33[医药卫生]