Department of Cardial Surgery, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(4):1797-1806. doi: 10.26355/eurrev_201902_17143.
The aim of this study was to explore the protective effect of nicotinamide on hypoxic cardiomyocytes and to investigate its possible mechanism.
Primary cardiomyocytes were used as study subjects. They were divided into three groups, including the blank group, control group and nicotinamide pretreatment group. Cell counting kit-8 (CCK-8) was used to detect cell viability. Lactate dehydrogenase (LDH) was used to detect cytotoxicity and flow cytometry was used to detect cell apoptosis. Polymerase Chain Reaction (PCR) and Western blot were used to measure the expressions of genes in adenosine monophosphate-activated protein kinase (AMPK) pathway. JC-1 detected the levels of mitochondrial membrane potential and reactive oxygen species (ROS). NAD was used for nicotinamide adenine dinucleotide (NAD)+, and NAD phosphate (NADP)+ levels. Adenosine triphosphate (ATP) assay was performed for the detection of intracellular energy metabolism.
In the absence of oxygen, nicotinamide had a protective effect on primary cardiomyocytes. Meanwhile, nicotinamide could markedly inhibit the increase of caspase3 mRNA in cardiomyocyte apoptosis pathway, and suppress the expression of apoptotic proteins. Furthermore, it could significantly induce the increase of intracellular ATP and activate the AMPK pathway. The detection of mitochondria indicated that nicotinamide alleviated hypoxic cardiomyocytes. In addition, the mitochondrial membrane potential disrupted and inhibited mitochondrial oxidative stress levels.
Nicotinamide pretreatment protects hypoxic cardiomyocytes and reduces intracellular mitochondrial stress. This protection may be related to the induction of the AMPK pathway and the increase of intracellular energy production.
本研究旨在探讨烟酰胺对缺氧心肌细胞的保护作用，并探讨其可能的机制。
原代心肌细胞作为研究对象。将其分为三组，空白组、对照组和烟酰胺预处理组。细胞计数试剂盒-8（CCK-8）用于检测细胞活力。乳酸脱氢酶（LDH）用于检测细胞毒性，流式细胞术用于检测细胞凋亡。聚合酶链反应（PCR）和蛋白质印迹用于测量腺苷单磷酸激活蛋白激酶（AMPK）通路中基因的表达。JC-1 检测线粒体膜电位和活性氧（ROS）水平。NAD 用于烟酰胺腺嘌呤二核苷酸（NAD+）和烟酰胺腺嘌呤二核苷酸磷酸（NADP+）水平。三磷酸腺苷（ATP）测定用于检测细胞内能量代谢。
在缺氧条件下，烟酰胺对原代心肌细胞具有保护作用。同时，烟酰胺可显著抑制心肌细胞凋亡途径中 caspase3mRNA 的增加，并抑制凋亡蛋白的表达。此外，它可以显著增加细胞内 ATP 的产生并激活 AMPK 通路。线粒体的检测表明烟酰胺减轻了缺氧心肌细胞的损伤。此外，线粒体膜电位破坏并抑制了线粒体氧化应激水平。
烟酰胺预处理可保护缺氧心肌细胞并减少细胞内线粒体应激。这种保护作用可能与诱导 AMPK 通路和增加细胞内能量产生有关。
