南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (7): 1127-1135.doi: 10.12122/j.issn.1673-4254.2023.07.09

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沉默生长抑制因子2可缓解血管紧张素Ⅱ诱导的小鼠心脏重塑：基于抑制p53乙酰化

刘正旺，邱晓堂，杨 华，吴小翠，叶文静   

  1. 海南省中医院心血管科，内分泌科，海南 海口 570203；广州中医药大学，广东 广州 510006
  • 出版日期:2023-07-20 发布日期:2023-07-20

Inhibitor of growth protein-2 silencing alleviates angiotensin II-induced cardiac remodeling in mice by reducing p53 acetylation

LIU Zhengwang, QIU Xiaotang, YANG Hua, WU Xiaocui, YE Wenjing   

  1. Department of Cardiovascular Medicine, Department of Endocrinology, Chinese Traditional Medicine Hospital of Hainan Province, Haikou 570203, China; Guangzhou University of Chinese Medicine, Guangzhou 510006, China
  • Online:2023-07-20 Published:2023-07-20

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摘要： 目的 明确Ing2对于血管紧张素 II（AngII）诱导的小鼠心脏重塑的影响和相关机制。方法 通过鼠尾静脉注射Ing shRNA2（Ad-sh-Ing2）腺病毒以沉默Ing2表达；通过持续微量注射泵注射Ang II（1000 ng·kg-1·min-1）诱导小鼠心脏重塑, 对照组给予相同体积生理盐水灌注。高频高分辨率超声成像技术评价各组小鼠心脏结构化功能及心脏肥大程度；马松和WGA染色检测小鼠心肌纤维化和心肌细胞横截面积；TUNEL染色检测心肌细胞凋亡水平，蛋白免疫印迹检测蛋白Cleaved-caspase3、Ing2、collagen Ⅰ、Ac- p53（Lys382）和p-p53（Ser15）表达；RT-qPCR检测Ing2 mRNA表达；线粒体ROS、ATP、柠檬酸合酶活性和线粒体钙储存试剂盒检测小鼠心肌组织线粒体功能。结果 相比于对照组，慢性AngII持续灌注小鼠的Ing2 mRNA和蛋白表达均明显增加（P<0.05）。慢性 Ang II灌注引起小鼠LVESD和LVEDD升高、LVEF和LVFS的下降（P<0.05）；Ing2沉默缓解了AngII诱发的小鼠心功能的减退，降低了LVESD和LVEDD，升高了LVEF和LVFS（P<0.05）。Ing2沉默能够改善 Ang II 诱导的心肌线粒体损伤，抑制心脏肥大、心肌纤维化和细心肌凋亡。慢性AngII持续灌注增加小鼠心肌组织中Ac- p53（Lys382）和p-p53（Ser15）水平，Ing2沉默能够抑制 Ang II 诱发Ac- p53（Lys382）的增加（P<0.05），但对p-p53（Ser15）磷酸化无显著影响。结论 Ing2可通过抑制p53乙酰化对抗AngII诱导的小鼠心脏重塑，抑制小鼠心功能的减退。

关键词: 血管紧张素 II；生长抑制因子2；p53；心肌肥厚；心脏重塑

Abstract: Objective To investigate the effect of inhibitor of growth protein-2 (Ing2) silencing on angiotensin II (AngII)-induced cardiac remodeling in mice and explore the underlying mechanism. Methods An adenoviral vector carrying Ing2 shRNA or empty adenoviral vector was injected into the tail vein of mice, followed 48 h later by infusion of 1000 ng · kg-1 · min-1 Ang II or saline using a mini-osmotic pump for 42 consecutive days. Transthoracic echocardiography was used to assess cardiac geometry and function and the level of cardiac hypertrophy in the mice. Masson and WGA staining were used to detect myocardial fibrosis and cross-sectional area of cardiomyocytes, and myocardial cell apoptosis was detected with TUNEL assay. Western blotting was performed to detect myocardial expressions of cleaved caspase 3, ING2, collagen Ⅰ, Ac-p53(Lys382) and p-p53 (Ser15); Ing2 mRNA expression was detected using real-time PCR. Mitochondrial biogenesis, as measured by mitochondrial ROS content, ATP content, citrate synthase activity and calcium storage, was determined using commercial assay kits. Results The expression levels of Ing2 mRNA and protein were significantly higher in the mice with chronic AngII infusion than in saline-infused mice. Chronic infusion of AngII significantly increased the left ventricular end-systolic diameter (LVESD) and left ventricular end-diastolic diameter (LVEDD) and reduced left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) in the mice. Ing2 silencing obviously alleviated AngII-induced cardiac function decline, as shown by decreased LVEDD and LVESD and increased LVEF and LVFS, improved myocardial mitochondrial damage and myocardial hypertrophy and fibrosis, and inhibited cardiomyocyte apoptosis. Chronic AngII infusion significantly increased myocardial expression levels of Ac-p53(Lys382) and p-p53(Ser15) in the mice, and Ing2 silencing prior to AngII infusion lessened AngII- induced increase of Ac-p53(Lys382) without affecting p53 (ser15) expression. Conclusion Ing2 silencing can inhibit AngII-induced cardiac remodeling and dysfunction in mice by reducing p53 acetylation.

Key words: angiotensin II; inhibitor of growth protein-2; P53; myocardial hypertrophy; cardiac remodeling