摘要
目的:研究花青素(Cyn)在压力超负荷诱导的小鼠心脏重构中的作用,并探讨其相关机制。方法:将120只6周龄雄性C57BL/6J小鼠随机分成4组:假手术(sham)组(n=20)、sham+Cyn组(n=20)、主动脉缩窄(TAC)组(n=40)和TAC+Cyn组(n=40)。采用TAC术建立小鼠慢性压力超负荷模型,造模当天记为第0天,sham+Cyn组和TAC+Cyn组提前5 d每日灌胃给予溶于二甲基亚砜及生理盐水的Cyn(5 mg·kg^-1·d^-1),sham组和TAC组提前5 d每日灌胃给予等量二甲基亚砜及生理盐水。TAC术后4周,分析比较各组的动物存活率,超声检测小鼠心功能,计算心重体重比和肺重体重比,麦胚凝集素染色和苏木素-伊红染色测量心肌细胞横截面积,通过二氢乙啶染色及超氧化物歧化酶(SOD)和丙二醛(MDA)检测来评估心脏氧化应激水平,TUNEL细胞凋亡检测试剂盒检测各组小鼠心肌细胞凋亡情况,RT-qPCR检测心房钠尿肽(ANP)、脑钠肽(BNP)和β-肌球蛋白重链(β-MHC)的mRNA表达情况,Western blot法检测Bax、Bcl-2、视神经萎缩蛋白1(OPA1)及发动蛋白相关蛋白1(Drp1)的蛋白表达。结果:与TAC组相比,Cyn干预可显著提高TAC后小鼠存活率(P<0.05),减少心肌细胞凋亡(P<0.05),减小心肌细胞横截面积(P<0.05),降低心重体重比和肺重体重比(P<0.05),降低活性氧与MDA含量(P<0.05),激活SOD(P<0.05)。M型超声检测显示,Cyn可显著提高TAC后小鼠的左心室射血分数和左心室短轴缩短率(P<0.05),同时减小左心室舒张期后壁厚度和左心室舒张末期内径(P<0.05)。透射电子显微镜结果表明,TAC后心肌线粒体数量增多且面积减小(P<0.05);Cyn干预后心肌线粒体面积增加且数量减少(P<0.05)。与sham组比较,TAC后OPA1蛋白表达显著降低(P<0.05);Cyn干预可增加TAC后OPA1蛋白的表达(P<0.05)。结论:Cyn显著提高TAC后小鼠存活率,并改善心脏功能,减轻心脏重构,其机制可能与Cyn抑制心肌线粒体OPA1剪切,促进线粒体融合有关。
AIM:To investigate the effect of cyanidin(Cyn)on pressure overload-induced cardiac remodeling and the underlying mechanism.METHODS:Six-week-old male C57BL/6 mice(n=120)were divided into 4 groups:sham group(n=20),sham+Cyn group(n=20),transverse aortic constriction(TAC)group(n=40)and TAC+Cyn group(n=40).The model of cardiac chronic pressure overload was induced by TAC,and the first day of TAC was defined as day 0.The animals in sham+Cyn group and TAC+Cyn group were treated with Cyn dissolved in DMSO and normal saline(5 mg·kg^-1·d^-1)for 5 d before TAC,while the animals in sham group and TAC group were treated with the same amount of DMSO and normal saline.Four weeks after TAC,the survival rate of the animals in each group was analyzed,the heart function of the mice was measured by ultrasound echocardiography,and the heart weight/body weight and lung weight/body weight were calculated.The cross-sectional area of the cardiomyocytes was measured by wheat germ agglutinin staining and hematoxylin-eosin staining.The degree of cardiac oxidative stress was evaluated by dihydroethidium staining and measurement of superoxide dismutase(SOD)and malondialdehyde(MDA)levels.The cardiomyocyte apoptosis was detected by TUNEL method.The mRNA expression levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP)andβ-myosin heavy chain(β-MHC)were detected by RT-qPCR,and the protein expression levels of Bax,Bcl-2,optic atrophy protein 1(OPA1)and dynamin-related protein 1(Drp1)were determined by Western blot.The mitochondrial morphological changes were observed by transmission electron microscopy.RESULTS:Compared with TAC group,the survival rate of the mice in TAC+Cyn group was significantly increased(P<0.05),the myocardial apoptosis,the cross-sectional area of myocardial cells,the heart weight/body weight,the lung weight/body weight,the level of reactive oxygen species and the MDA content were decreased(P<0.05),and the SOD was activated(P<0.05).M-mode ultrasound tests showed that Cyn treatment significantly increased l
作者
曾静静
徐肖肖
施涵青
徐光宇
金其可
阮永雪
任芳芳
谢作益
李磊
ZENG Jing-jing;XU Xiao-xiao;SHI Han-qing;XU Guang-yu;JIN Qi-ke;RUAN Yong-xue;REN Fang-fang;XIE Zuo-yi;LI Lei(Department of Cardiology,The Second Affiliated Hospital&Yuying Children's Hospital of Wenzhou Medical University,Wenzhou 325027,China;Enze Hospital,Taizhou Enze Medical Center Group,Taizhou 318050,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第12期2113-2122,共10页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81570238,No.81770292)
贺林院士工作站科研基金资助项目(No.18331101)。
关键词
花青素
主动脉缩窄
心脏重构
视神经萎缩蛋白1
线粒体融合
Cyanidin
Transverse aortic constriction
Cardiac remodeling
Optic atrophy protein 1
Mitochondrial fusion
