天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (1): 38-44.doi: 10.11958/20190267

• 实验研究 • 上一篇    下一篇

Akt-Kv4.3-CaMKⅡ在有氧运动抑制压力负荷小鼠 心肌肥厚中的作用及其机制

葛广全 1，赵峰 2，陈道虎 1，石振塑 1，陈泽伦 1，王天光 1，何书武 1，魏以桢 3△   

  1. 基金项目：海南省卫生厅科研课题（1521320273A2001） 作者单位：1海口，海南医学院第二附属医院心血管外科（邮编570311）；2武汉，华中科技大学同济医学院附属协和医院外科；3北京，中国 医学科学院阜外医院外科 作者简介：葛广全（1972），男，本科，副主任医师，主要从事心衰及心梗发病机制研究 △通讯作者 E-mail: weiyizhen@sohu.com
  • 收稿日期:2019-01-25 修回日期:2019-07-22 出版日期:2020-01-15 发布日期:2020-01-15
  • 通讯作者: 葛广全 E-mail:1660204443@qq.com
  • 基金资助:
    海南省卫生厅科研课题

The role and mechanism of Akt-Kv4.3-CaMKII in aerobic exercise inhibited pressure overload-induced cardiac hypertrophy

GE Guang-quan1, ZHAO Feng2, CHEN Dao-hu1, SHI Zhen-su1, CHEN Ze-lun1, WANG Tian-guang1, HE Shu-wu1, WEI Yi-zhen3△   

  1. 1 Department of Cardiovascular Surgery, the Second Affiliated Hospital, Hainan Medical College, Haikou 570311, China; 2 Department of Hand Surgery, Wuhan Union Hospital, Tongji Medical College, Huazhong University of Science and Technology; 3 Department of Surgery, Fuwai Hospital, Chinese Academy of Medical Sciences △Corresponding Author E-mail: weiyizhen@sohu.com
  • Received:2019-01-25 Revised:2019-07-22 Published:2020-01-15 Online:2020-01-15

PDF (PC)

4711

摘要： 目的 探讨丝氨酸/苏氨酸激酶（Akt）-离子通道Kv4.3（Kv4.3）-钙调素依赖性蛋白激酶Ⅱ（CaMKⅡ）信号 通路在有氧运动抑制压力负荷小鼠心肌肥厚中的作用及其机制。方法 60只实验小鼠均分为5组：假手术（SHAM） 组、主动脉缩窄手术（TAC）组、假手术+有氧运动（SHAM+E）组、主动脉缩窄术+有氧运动（TAC+E）组，主动脉缩窄术+ 有氧运动+ Akt抑制剂Perifosine（TAC+E+Peri）组。高分辨率小动物超声系统评价小鼠心功能及肥厚程度，麦胚凝集 素（WGA）染色检测心肌细胞横截面积；荧光定量PCR（RT-qPCR）检测ANP表达，Western blot检测蛋白心房钠尿肽 （ANP）、p-Akt、Kv4.3、p-CaMKⅡ表达水平。结果 与SHAM组相比，TAC促进心肌肥厚，恶化心功能（P＜0.01）。给 予有氧运动训练后，TAC+E组较TAC组小鼠心功能改善，心肌肥厚程度减轻（P＜0.01）。同时Western blot检测显示， TAC组较SHAM组p-Akt、Kv4.3表达降低，p-CaMKⅡ上调（P＜0.01）；TAC+E组p-Akt和Kv4.3表达较TAC小鼠增加， p-CaMKⅡ下调（P＜0.01）。而给予Akt抑制剂Perifosine后，相比于TAC+E组，TAC+E+Peri组心功能恶化、心肌肥厚 程度和ANP表达增加，心肌细胞横截面积增大，同时伴随Kv4.3表达降低、p-CaMKⅡ增高（P＜0.01）。结论 有氧运 动训练能够通过调节Akt-Kv4.3-CaMKⅡ信号通路抑制压力负荷心肌肥厚。

关键词: 心肌病, 肥厚性, 蛋白质丝氨酸苏氨酸激酶, 钙-钙调素依赖性蛋白激酶2型, 疾病模型, 动物, 有氧运动, 离子通道Kv4.3

Abstract: Objective To investigate the role and mechanism of serine / threonine kinase (Akt) - ion channel Kv4.3 (Kv4.3) - calmodulin-dependent protein kinase Ⅱ (CaMK Ⅱ) signaling pathway in aerobic exercise inhibited pressure overload-induced cardiac hypertrophy. Methods Sixty mice were divided into five groups: Sham group, transverse aortic constriction (TAC) group, SHAM+ aerobic exercise (E) group, TAC + E group and TAC+ E + AKt inhibitor perifosine (Peri) group. Transthoracic echocardiography was used to assess the cardiac function and extent of myocardial hypertrophy. The cross-sectional area of myocardial cells was measured by WGA staining. The mRNA expression of ANP was detected by RTqPCR. The protein expression levels of ANP, p-AKt, Kv4.3 and p-CaMKⅡ were detected by Western-blot assay. Results Compared with Sham group, the cardiac function of mice was deteriorated in TAC group (P＜0.01), and the extent of cardiac hypertrophy was increased (P＜0.01). After aerobic exercise training, the cardiac function was improved in TAC + E group, and the level of myocardial hypertrophy was alleviated compared with TAC group (P＜0.01). Western-blot assay showed that the expressions of p-AKT and Kv4.3 were down-regulated, p-CaMKⅡ was up-regulated in TAC group than those in Sham group (P＜0.01). However, the expressions of p-AKT and Kv4.3 were higher, p-CaMKⅡ was lower in TAC+E group than those in TAC mice (P＜0.01). Furthermore, pretreatment with the AKT inhibitor perifosine, deteriorated cardiac function, augmented myocardial hypertrophy and ANP, increased cross-sectional area were observed in TAC+E+Peri group compared with those of TAC+E group (P＜0.01). Meanwhile, the downregulation of p-AKT、Kv4.3 and upregulation p-CaMKⅡ were detected in the TAC+E+Peri group compared with the TAC+E group. Conclusion Aerobic exercise training can inhibit pressure overload-induced cardiac hypertrophy by regulating Akt-Kv4.3-CaMKⅡ.

Key words: cardiomyopathy, hypertrophic, protein-serine-threonine kinases, calcium-calmodulin-dependent protein kinase type 2, disease models, animal, aerobic exercise, Kv4.3