Ghrelin介导HO-1/PGC-1α信号通路调节线粒体氧化应激改善大鼠创伤性脑损伤  

Ghrelin-mediated HO-1/PGC-1αsignaling pathway regulates mitochondrial oxidative stress to improve traumatic brain injury in rats

作　　者：赵智慧 翟秀丽 王晶 马敏 白香花 苏楠 ZHAO Zhihui;ZHAI Xiuli;WANG Jing;MA Min;BAI Xianghua;SU Nan(Department of Anesthesiology,Inner Mongolia People’s Hospital,Hohhot 010017,China)

机构地区：[1]内蒙古自治区人民医院麻醉科,呼和浩特010017

出　　处：《中国医科大学学报》2025年第4期351-358,共8页Journal of China Medical University

基　　金：内蒙古自然科学基金(2023LHMS08048);内蒙古自治区人民医院院内基金项目(2021YN01);内蒙古医学科学院公立医院科研联合基金科技项目(2023GLLH0025)。

摘　　要：目的基于HO-1/PGC-1α信号通路探讨Ghrelin对大鼠创伤性脑损伤的保护作用。方法SPF级雄性大鼠30只,随机分为假手术组、创伤性脑损伤(TBI)组以及Ghrelin干预(Ghrelin)组,每组10只。采用Feeney自由落体撞击法制备大鼠TBI模型,Ghrelin组在造模后30 min尾静脉注射Ghrelin(20μg/kg),假手术组不做撞击。造模72 h后取大鼠脑组织,测量各组大鼠脑含水量来分析脑水肿严重程度;HE染色观察各组大鼠脑组织病理变化;ELISA检测各组大鼠脑组织氧化应激因子MDA、SOD和GSH-Px水平;TUNEL染色检测各组大鼠脑组织细胞凋亡;Western blotting检测各组大鼠脑组织Bcl-2、Bax、caspase-3和caspase-9蛋白表达水平;免疫荧光检测各组大鼠脑组织线粒体活性氧(mtROS),Western blotting检测线粒体分裂素Mfn1/2、核呼吸因子1(NRF1)和线粒体转录因子A(TFAM)蛋白表达水平;Western blotting检测各组大鼠脑组织HO-1、PGC-1α蛋白表达水平。20只Ghrelin干预的TBI模型大鼠分为Ghrelin+sh-NC组和Ghrelin+sh-HMOX1组,每组10只;大鼠在给予Ghrelin干预的同时经尾静脉注射敲减对照(腺病毒2.5×10^(9) pfu)或敲减HMOX1腺病毒(2.5×10^(9) pfu)。Western blotting检测2组大鼠脑组织中HO-1、PGC-1α、Bcl-2、Bax、caspase-3和caspase-9蛋白表达;ELISA检测2组大鼠脑组织MDA、SOD和GSH-Px水平。结果与假手术组比较,TBI组大鼠脑组织病理损伤和脑水肿加重,脑细胞凋亡数量增多,氧化应激因子SOD、GSH-Px水平降低,MDA水平升高,脑组织mtROS升高,Bcl-2蛋白表达减少,Bax、caspase-3和caspase-9蛋白表达增加,Mfn1/2、NRF1、TFAM、HO-1和PGC-1α蛋白表达水平降低(均P<0.05)。与TBI组比较,Ghrelin组大鼠脑组织病理损伤改善,脑水肿减轻,脑细胞凋亡数量减少,氧化应激因子SOD、GSH-Px水平升高,MDA水平降低,脑组织mtROS降低,Bcl-2蛋白表达增加,Bax、caspase-3和caspase-9蛋白表达降低,Mfn1/2、NRF1、TFAM、HO-1和PGC-1α蛋白表达增加(均P<0.05)。与Objective To investigate the protective effect of Ghrelin on traumatic brain injury(TBI)in rats based on the HO-1/PGC-1αsignaling pathway.Methods Thirty SPF male rats were randomly divided into sham,TBI,and Ghrelin groups,with 10 rats in each group.A TBI rat model was established using the Feeney free-fall impingement method.The Ghrelin group was injected by caudal vein at a dose of 20μg/kg 30 min after modeling,while the sham group was not impinged.After 72 h of modeling,the brain tissues of the rats were collected,and the brain water content was measured in order to analyze the severity of brain edema.HE staining was used to observe pathological changes in brain tissue.The levels of the oxidative stress factors MDA,SOD,and GSH-Px were determined using ELISA.TUNEL staining was used to detect the apoptosis of the brain cells,and the expression levels of Bcl-2,Bax,caspase-3 and caspase-9 in the brain tissues were detected by Western blotting.Mitochondrial reactive oxygen species(mtROS)in the brain tissues were detected by immunofluorescence.Mitochondrial function indicators,including mitochondrial mitogen Mfn1/2,nuclear respiration factor 1(NRF1),and mitochondrial transcription factor A(TFAM)were detected by Western blotting.The expression levels of HO-1 and PGC-1αin the brain tissues of rats in each group were detected by Western blotting.Twenty TBI model rats treated with Ghrelin were divided into Ghrelin+sh-NC and Ghrelin+sh-HMOX1 group with 10 rats in each group.Rats were treated with Ghrelin and injected with knock-down control(adenovirus 2.5×10^(9) pfu)or knock-down HMOX1 adenovirus(2.5×10^(9) pfu)via tail vein.Western blotting was used to detect the expressions of HO-1,PGC-1α,Bcl-2,Bax,caspase-3 and caspase-9 in the brain tissues of the two groups.The levels of MDA,SOD and GSH-Px in brain tissue of two groups were detected by ELISA.Results Compared with the sham group,the pathological injury and brain edema in TBI group were aggravated,the number of brain cell apoptosis increased,the levels of oxidati

关 键 词：GHRELIN 创伤性脑损伤 大鼠 HO-1/PGC-1α信号通路 氧化应激 

分 类 号：R651.15[医药卫生—外科学]