烟酰胺磷酸核糖转移酶在小鼠心肌梗死后修复中的表达变化与功能研究  

Expression Changes and Function of Nicotinamide Phosphoribosyltransferase in Post-infarction Repair in Mice

作　　者：钟嘉俊 黄成珍 聂宇[3] 林爱玲[1] 王珏[1] ZHONG Jiajun;HUANG Chengzhen;NIE Yu;LIN Ailing;WANG Jue(Department of Cardiac Surgery,The First Affiliated Hospital of Wenzhou Medical University,Wenzhou 325000,China;Institute of Basic Medical Sciences of Chinese Academy of Medical Sciences,School of Basic Medicine of Peking Union Medical College,Beijing 100005,China;State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases and Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,China)

机构地区：[1]温州医科大学附属第一医院心脏外科,温州325000 [2]中国医学科学院基础医学研究所北京协和医学院基础学院,北京100005 [3]中国医学科学院北京协和医学院国家心血管病中心阜外医院心血管疾病国家重点实验室,北京100037现为温州医科

出　　处：《中国循环杂志》2023年第4期456-463,共8页Chinese Circulation Journal

基　　金：2021年浙江省医坛新秀(浙卫办[2021]40号);温州市科学技术局自筹课题(2021Y1307)。

摘　　要：目的:探讨心肌梗死后烟酰胺磷酸核糖转移酶(NAMPT)表达变化及其对小鼠心肌细胞增殖的影响,为治疗心肌梗死探索新的理论依据与干预靶点。方法:分离原代乳鼠心肌细胞,给予NAMPT重组蛋白100 ng/ml,利用免疫荧光染色检测心肌细胞的磷酸化组蛋白H3(pH3)、增殖抗原Ki67表达。将成年C57BL/6j雄性小鼠40只随机分成2组:NAMPT治疗组,磷酸盐缓冲液(PBS)对照组,每组20只,同时取10只作为假手术组。通过结扎小鼠冠状动脉左前降支建立小鼠心肌梗死模型,NAMPT治疗组于结扎线以下通过显微心肌内注射NAMPT重组蛋白0.5μg,PBS对照组以同样方法注射PBS溶液,假手术组仅开胸不结扎冠状动脉。通过M型超声检测小鼠术后心功能,取出心脏后Masson染色在光学显微镜下观察并测定心肌梗死面积。结果:左前降支结扎构建心肌梗死模型的小鼠在术后第2、3天NAMPT的信使RNA(mRNA)均显著下调(P均<0.05)。NAMPT处理原代心肌细胞后,与PBS对照组相比,pH3、Ki67阳性心肌细胞比例差异均无统计学意义(P均>0.05),心肌内注射NAMPT的乳鼠心脏中pH3阳性的心肌细胞较PBS对照组也无明显增多(P>0.05)。心肌梗死小鼠NAMPT给药后,与PBS对照组相比,NAMPT治疗组的左心室射血分数和左心室缩短分数差异均无统计学意义(P均>0.05),NAMPT治疗组的心重、体重、心重/体重比值差异均无统计学意义(P均>0.05);两组间梗死面积差异无统计学意义(P>0.05)。结论:心肌梗死损伤会导致心肌组织中NAMPT表达水平下调;但外源性补给NAMPT不能改善小鼠心肌梗死后的心功能,不能减小梗死面积,NAMPT在心肌梗死损伤修复中的作用有待进一步探究。Objectives:To investigate nicotinamide phosphoribosyltransferase(NAMPT)expression changes post myocardial infarction and its effect on cardiomyocyte proliferation in mice,and to explore new theoretical basis and intervention targets for the treatment of myocardial infarction.Methods:Primary neonatal mouse cardiomyocytes were isolated and treated with NAMPT recombinant protein(100 ng/ml),and the phosphorylated histone H3(pH3)and Ki67 expression in cardiomyocytes were detected using immunofluorescence staining.Forty male adult C57BL/6j mice were randomly divided into 2 groups:NAMPT treatment group,phosphate buffer saline(PBS)control group(n=20 each),and a sham operation group(n=10)was also established.Myocardial infarction(MI)mice model was established by ligating the anterior descending branch of the left coronary artery.The postoperative cardiac function of the mice was detected by M-mode echocardiography,and Masson-stained myocardial sections were observed under light microscope to determine the area of myocardial infarction.Results:The mRNA expression of NAMPT was significantly downregulated in MI mice on day 2 and 3 post infarction(both P<0.05).In vitro,pH3 and Ki67-positive cardiomyocytes were not significantly different after NAMPT treatment compared with the PBS control group(both P>0.05).There was no significant increase in the number of pH3-positive cardiomyocytes in the hearts of neonatal mice injected with NAMPT compared to the PBS control group(P>0.05).There were no significant differences in left ventricular ejection fraction,left ventricular shortening fraction,heart weight,body weight,heart weight to body weight ratio,and infarct size between the NAMPT treatment group(intramyocardial injection)and the PBS control group(all P>0.05).Conclusions:Myocardial infarction leads to down-regulation of NAMPT expression level in myocardial tissue,but exogenous supplementation of NAMPT does not affect cardiac function and infarct area in mice after infarction,and the role of NAMPT in myocardial infarction repair

关 键 词：心肌梗死 心肌增殖 烟酰胺磷酸核糖转移酶 心肌内注射 

分 类 号：R54[医药卫生—心血管疾病]