摘要
背景:对比剂肾病已经成为医院获得性急性肾损伤的主要原因之一,其本质是肾脏的急性缺血再灌注损伤,其中去乙酰化酶3在其中发挥的作用还不清楚。目的:探索不同肾脏缺血时间对去乙酰化酶3表达的影响及其与线粒体损伤相关指标的关系。方法:构建C57BL/6J小鼠急性肾脏缺血模型,给予不同的缺血时间(15,20,25,30 min),再灌注48 h。根据缺血时间,分为对照组、假手术组、缺血15,20,25,30 min再灌注组,每组8只。术后48 h,检测血清肌酐、尿素氮水平,TUNEL试剂盒检测肾组织细胞凋亡情况,荧光素酶发光法检测肾组织ATP水平,苏木精-伊红染色观察肾组织病理变化,透射电镜下观察线粒体变化,Western blot检测去乙酰化酶3和线粒体动力相关蛋白1、线粒体融合蛋白1的表达。结果与结论:①血清肌酐、尿素氮、组织病理评分及凋亡指数在各缺血组中逐渐升高;②肾组织线粒体结构损伤在各缺血组中逐渐加重,ATP含量总体下降;③正常对照组与假手术组及缺血15min再灌注组的去乙酰化酶3蛋白水平无差异;与缺血15min再灌注组比较,缺血20 min再灌注组的去乙酰化酶3蛋白水平升高(P<0.05),缺血25 min继续升高(P<0.05),缺血30 min表达最高(P<0.05);④与假手术组比较,线粒体融合蛋白1在缺血15-20 min升高(P<0.05);与缺血15 min再灌注组比较,线粒体融合蛋白1在缺血25 min和30 min进一步升高(P<0.05);⑤缺血15 min再灌注组的线粒体动力相关蛋白1水平达高峰,与缺血15 min再灌注组比较,缺血20 min再灌注组的线粒体动力相关蛋白1水平有所下降(P<0.05),缺血25 min再灌注组进一步下降(P<0.05);⑥相关性研究发现,ATP与去乙酰化酶3存在显著负相关(r=-0.77,P<0.05),去乙酰化酶3与线粒体动力相关蛋白1存在负向相关性(r=-0.52,P<0.05),去乙酰化酶3与线粒体融合蛋白1存在正向相关性(r=0.72,P<0.05);⑦结果表明,肾脏缺血再灌注损伤时,ATP水�
BACKGROUND:Contrast-induced nephropathy has become one of the main causes of hospital-acquired acute kidney injury and its essence is acute renal ischemia-reperfusion injury.However,the role of sirtuin 3 in it is still unclear.OBJECTIVE:To probe the dynamical changes of sirtuin 3 after different duration of renal ischemia and to establish the relationship of sirtuin3 expression with mitochondrial injury-related parameters.METHODS:The mouse model of acute renal ischemia-reperfusion injury was established by ischemia for different time(15,20,25,30 minutes)followed by 48 hours of reperfusion in C57BL/6 mice.According to the ischemic time,all animals were divided into six groups(n=8 per group):control group,sham group,15-,20-,25-and 30-minute ischemia groups.Forty-eight hours after modeling,the serum levels of creatinine and blood urea nitrogen were measured.The apoptosis of renal tissue cells was monitored by TUNEL.Adenosine triphosphate content in the kidney was measured by luciferase luminescence method.Hematoxylin-eosin staining was used to observe pathological changes of the kidney.Mitochondrial changes were observed under a transmission electron microscope.The expression of mitochondrial dynamin related protein 1,mitofusion 1,and sirtuin 3 was determined by western blot.RESULTS AND CONCLUSION:After ischemia,serum ccreatinine,blood urea nitrogen,histopathological score and apoptotic index increased gradually after ischemia;the injury of mitochondrial structure of the kidney gradually aggravated and the mitochondrial adenosine triphosphate content decreased in general.The sirtuin 3 expression showed no significant difference among control,sham and 15-minute ischemia groups.The expression of sirtuin 3 was higher at 20minutes of ischemia than at 15 minutes of ischemia(P<0.05),further increased significantly at 25 minutes of ischemia(P<0.05),and reached a peak at 30minutes of ischemia(P<0.05).Mitofusion 1 expression significantly increased in the 15-and 20-minute ischemia groups compared with the sham group(P<0.05)a
作者
贾圣琪
罗文龙
田丁元
张新会
崔茜
王超
裴汉军
Jia Shengqi;Luo Wenlong;Tian Dingyuan;Zhang Xinhui;Cui Qian;Wang Chao;Pei Hanjun(Department of Cardiology,The First Affiliated Hospital,Baotou Medical College of Inner Mongolia University of Science and Technology,Baotou 014010,InnerMongolia Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2023年第8期1172-1178,共7页
Chinese Journal of Tissue Engineering Research
基金
内蒙古自治区科技计划项目(201702101),项目负责人:裴汉军
内蒙古自治区高等学校“青年科技英才支持计划”(NJYT-17-A20),项目负责人:裴汉军
国家自然科学基金(81760096),项目负责人:裴汉军。
