利拉鲁肽可能通过调节细胞内钙稳态直接保护心肌细胞免受再灌注损伤。

Liraglutide directly protects cardiomyocytes against reperfusion injury possibly via modulation of intracellular calcium homeostasis.

作者信息

Hu Shun-Ying, Zhang Ying, Zhu Ping-Jun, Zhou Hao, Chen Yun-Dai

机构信息

Department of Cardiology, Chinese PLA General Hospital, Beijing, China.

出版信息

J Geriatr Cardiol. 2017 Jan;14(1):57-66. doi: 10.11909/j.issn.1671-5411.2017.01.008.

BACKGROUND

Liraglutide is glucagon-like peptide-1 receptor agonist for treating patients with type 2 diabetes mellitus. Our previous studies have demonstrated that liraglutide protects cardiac function through improving endothelial function in patients with acute myocardial infarction undergoing percutaneous coronary intervention. The present study will investigate whether liraglutide can perform direct protective effects on cardiomyocytes against reperfusion injury.

METHODS

experiments were performed using H9C2 cells and neonatal rat ventricular cadiomyocytes undergoing simulative hypoxia/reoxygenation (H/R) induction. Cardiomyocytes apoptosis was detected by fluorescence TUNEL. Mitochondrial membrane potential (Ψm) and intracellular reactive oxygen species (ROS) was assessed by JC-1 and DHE, respectively. Fura-2/AM was used to measure intracellular Ca concentration and calcium transient. Immunofluorescence staining was used to assess the expression level of sarcoplasmic reticulum Ca-ATPase (SERCA2a). experiments, myocardial apoptosis and expression of SERCA2a were detected by colorimetric TUNEL and by immunofluorescence staining, respectively.

RESULTS

liraglutide inhibited cardiomyotes apoptosis against H/R. Ψm of cardiomyocytes was higher in liraglutide group than H/R group. H/R increased ROS production in H9C2 cells which was attenuated by liraglutide. Liraglutide significantly lowered Ca overload and improved calcium transient compared with H/R group. Immunofluorescence staining results showed liraglutide promoted SERCA2a expression which was decreased in H/R group. In ischemia/reperfusion rat hearts, apoptosis was significantly attenuated and SERCA2a expression was increased by liraglutide compared with H/R group.

CONCLUSIONS

Liraglutide can directly protect cardiomyocytes against reperfusion injury which is possibly through modulation of intracellular calcium homeostasis.

背景

利拉鲁肽是一种用于治疗2型糖尿病患者的胰高血糖素样肽-1受体激动剂。我们之前的研究表明，利拉鲁肽通过改善接受经皮冠状动脉介入治疗的急性心肌梗死患者的内皮功能来保护心脏功能。本研究将调查利拉鲁肽是否能对心肌细胞产生直接保护作用以抵抗再灌注损伤。

方法

使用H9C2细胞和经历模拟缺氧/复氧（H/R）诱导的新生大鼠心室心肌细胞进行实验。通过荧光TUNEL检测心肌细胞凋亡。分别使用JC-1和DHE评估线粒体膜电位（Ψm）和细胞内活性氧（ROS）。使用Fura-2/AM测量细胞内钙浓度和钙瞬变。免疫荧光染色用于评估肌浆网Ca-ATP酶（SERCA2a）的表达水平。在实验中，分别通过比色法TUNEL和免疫荧光染色检测心肌凋亡和SERCA2a的表达。

结果

利拉鲁肽抑制心肌细胞针对H/R的凋亡。利拉鲁肽组心肌细胞的Ψm高于H/R组。H/R增加了H9C2细胞中的ROS产生，而利拉鲁肽可使其减弱。与H/R组相比，利拉鲁肽显著降低了钙超载并改善了钙瞬变。免疫荧光染色结果显示，利拉鲁肽促进了SERCA2a的表达，而SERCA2a在H/R组中减少。在缺血/再灌注大鼠心脏中，与H/R组相比，利拉鲁肽显著减轻了凋亡并增加了SERCA2a的表达。