过氧化物酶体增殖物激活受体γ共激活因子-1α在瑞芬太尼抗肝脏缺血-再灌注损伤中的作用研究  

Study on the role of peroxisome proliferator-activated receptorγcoactivator-1αin remifentanil against hepatic ischemia-reperfusion injury

作　　者：陈玲利 李秀芳 郝泉水 张喜华 Chen Lingli;Li Xiufang;Hao Quanshui;Zhang Xihua(Department of Anesthesiology,Huangzhou District People's Hospital in Huanggang City,Huanggang 438000,China;不详)

机构地区：[1]黄冈市黄州区人民医院麻醉科,湖北黄冈438000 [2]黄冈市中心医院麻醉科 [3]黄冈市中心医院肝胆外科

出　　处：《器官移植》2025年第2期246-255,共10页Organ Transplantation

基　　金：湖北省卫生健康委科研项目(WJ2021M084、WJ2023M176);湖南省人民医院医联体专项科研基金项目(2022YLT006)。

摘　　要：目的探讨过氧化物酶体增殖物激活受体γ共激活因子-1α(PGC-1α)在瑞芬太尼(REM)抗肝脏缺血-再灌注损伤(HIRI)中的作用及其机制。方法将40只SD大鼠随机分为假手术(sham)组、HIRI组、HIRI+REM组、HIRI+PGC-1α抑制剂SR-18292(HIRI+SR-18292)组和HIRI+REM+SR-18292组,每组8只。采用无创动脉夹阻断法构建HIRI大鼠模型,并于术前分别静脉注射REM或SR-18292。使用试剂盒检测大鼠血清中肝功能指标及肝组织三磷酸腺苷(ATP)水平;比色法检测大鼠肝组织中线粒体呼吸链复合物Ⅲ、Ⅳ(COX-Ⅲ、COX-Ⅳ)活性水平;苏木素-伊红染色观察大鼠肝组织病理学改变;荧光探针(DCFH-DA)法和比色法检测大鼠肝组织中活性氧簇(ROS)及氧化应激相关指标水平;实时荧光定量聚合酶链反应(RT-qPCR)检测大鼠肝组织中线粒体DNA(mtDNA)拷贝数及PGC-1α、核呼吸因子-1(NRF-1)和线粒体转录因子A(TFAM)信使RNA(mRNA)表达水平;蛋白质印迹法检测大鼠肝组织中PGC-1α、NRF-1和TFAM蛋白表达水平。结果与sham组比较,HIRI组大鼠肝组织病理评分及肝组织细胞坏死程度增加,血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平和肝组织中ROS、丙二醛(MDA)水平升高,肝组织中ATP含量以及超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、COX-Ⅲ和COX-Ⅳ活性水平降低,肝组织中mtDNA拷贝数以及PGC-1α、NRF-1、TFAM mRNA和蛋白表达水平降低(均为P<0.05)。与HIRI组比较,HIRI+REM组大鼠肝组织病理评分及肝组织细胞坏死程度下降,血清中ALT、AST水平和肝组织中ROS、MDA水平降低,肝组织中ATP含量以及SOD、GSH-Px、COX-Ⅲ和COX-Ⅳ活性水平升高,肝组织中mtDNA拷贝数以及PGC-1α、NRF-1、TFAM mRNA和蛋白表达水平升高(均为P<0.05),而HIRI+SR-18292组大鼠肝组织病理评分及肝组织细胞坏死程度增加,血清中ALT、AST水平和肝组织中ROS、MDA水平升高,肝组织中ATP含量以及SOD、GSHPx、COX-Ⅲ和COX-Ⅳ活�Objective To explore the role and mechanism of peroxisome proliferator-activated receptorγcoactivator-1α(PGC-1α)in remifentanil(REM)resistance to hepatic ischemia-reperfusion injury(HIRI).Methods Forty SD rats were randomly divided into sham surgery group,HIRI group,HIRI+REM group,HIRI+PGC-1αinhibitor SR-18292(HIRI+SR-18292)group and HIRI+REM+SR-18292 group,8 rats in each group.HIRI rat models were constructed using non-invasive arterial clip occlusion method,and REM or SR-18292 were intravenously injected before surgery.The liver function indicators and liver tissue adenosine triphosphate(ATP)levels in the serum of rats were detected by assay kits.The activity levels of mitochondrial respiratory chain complexesⅢandⅣ(COX-Ⅲ,COX-Ⅳ)in rat liver tissue were assessed by colorimetric methods.The pathological changes in rat liver tissue were observed by hematoxylin-eosin staining.Reactive oxygen species(ROS)and oxidative stress-related indicators in rat liver tissue were measured using the fluorescent probe(DCFH-DA)method and colorimetric methods.The mitochondrial DNA(mtDNA)copies and the expression levels of PGC-1α,nuclear respiratory factor-1(NRF-1)and mitochondrial transcription factor A(TFAM)messenger RNA(mRNA)in rat liver tissue were quantified by real-time fluorescent quantitative polymerase chain reaction(RT-qPCR).And the protein expression levels of PGC-1α,NRF-1 and TFAM in rat liver tissue were assessed by Western blotting.Results Compared with the sham group,rats in the HIRI group showed increased pathological scores and hepatic cell necrosis in liver tissue,elevated levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum,and increased levels of ROS and malondialdehyde(MDA)in liver tissue.Additionally,there was a decrease in ATP content and the activity levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),COX-Ⅲand COX-Ⅳin liver tissue,as well as a decrease in mtDNA copies and the expression levels of PGC-1α,NRF-1 and TFAM mRNA and protein(all P<0.05).

关 键 词：肝脏缺血-再灌注损伤 瑞芬太尼 过氧化物酶体增殖物激活受体γ共激活因子-1α 线粒体呼吸链复合物 活性氧簇 线粒体DNA 线粒体转录因子A 核呼吸因子-1 

分 类 号：R617[医药卫生—外科学] R971[医药卫生—临床医学]