心肌缺血再灌注(IR)损伤是由于心脏血流的最初减少而引起的损伤,阻止它接受足够的氧气,以及随后通过阻塞的冠状动脉的开口恢复血流,产生矛盾的有害影响。发现预防IR的新疗法至关重要。大蒜素是一种从大蒜中分离出来的化合物,具有预防和治疗不同疾病的能力,对心肌也有保护作用。因此,本研究旨在评价大蒜素对心肌细胞IR损伤的保护作用。
我们建立了原代猪心肌细胞的体外缺氧-复氧(HR)模型,以模拟心肌IR损伤。从小型麝香猪(1日龄)中提取原代猪心肌细胞。经过至少2-3天的适应期,选取状况良好的心肌细胞,随机分为对照组(常氧持续5h),HR组(缺氧2h/复氧3h),和HR+大蒜素组(缺氧/复氧+大蒜素治疗)。
诱导缺氧/复氧后,大蒜素处理增强了细胞活力。此外,与HR组相比,大蒜素处理导致细胞凋亡从13.5±1.2%减少到6.11±0.15%(p<0.05),细胞凋亡相关蛋白也受到调控,Bax的表达减少,在HR+大蒜素组中,caspase-3和胞质细胞色素C裂解,Bcl-2表达增加(均p<0.01)。促炎细胞因子,如白细胞介素-6和肿瘤坏死因子α被大蒜素治疗下调(均P<0.01)。此外,显著降低细胞内活性氧的产生(p<0.01),减少线粒体膜电位的损失(p<0.01)。此外,PPARγ共激活因子-1α和内皮型一氧化氮合酶的表达上调(p均<0.01),大蒜素处理可下调内皮素-1、缺氧诱导因子-1α和转化生长因子β的表达(均p<0.01)。
这些结果表明大蒜素通过减少细胞凋亡保护心肌细胞免受HR损伤,炎症和线粒体损伤,从而为其在心肌IR治疗中的潜在应用提供了依据。
Myocardial ischemia-reperfusion (IR) injury is a damage due to an initial reduction in blood flow to the heart, preventing it from receiving enough oxygen, and subsequent restoration of blood flow through the opening of an occluded coronary artery producing paradoxical harmful effects. The finding of new therapies to prevent IR is of utmost importance. Allicin is a compound isolated from garlic having the ability to prevent and cure different diseases, and a protective effect on the myocardium was also demonstrated. Therefore, the aim of this
study was to evaluate the in vitro protective effect of Allicin against myocardial IR injury on cardiomyocytes.
We established an in vitro hypoxia-reoxygenation (HR) model of primary porcine cardiomyocytes to simulate myocardial IR injury. Primary porcine cardiomyocytes were extracted from Mini-musk swines (1 day old). After a period of adaptation of at least 2-3 days, cardiomyocytes in good condition were selected and randomly divided into control group (normal oxygen for 5 h), HR group (2 h of hypoxia/3 h of reoxygenation), and HR + Allicin group (hypoxia/reoxygenation + Allicin treatment).
After the induction of hypoxia/reoxygenation, Allicin treatment enhanced the cell viability. Moreover, Allicin treatment resulted in a reduction of apoptosis from 13.5 ± 1.2% to 6.11 ± 0.15% compared with the HR group (p < 0.05), and the apoptosis related proteins were regulated as well, with a decreased expression of Bax, cleaved caspase-3 and cytosolic cytochrome C and an increase in Bcl-2 expression in the HR + Allicin group (all p < 0.01). Pro-inflammatory cytokines, such as interleukin-6 and tumor necrosis factor alpha were down-regulated by the treatment with Allicin (both p < 0.01). In addition, it significantly decreased intracellular reactive oxygen species generation (p < 0.01) and reduced the loss of mitochondrial membrane potential (p < 0.01). Furthermore, the expression of PPARγ coactivator-1α and endothelial nitric oxide synthase was up-regulated (both p < 0.01), while the expression of Endothelin-1, hypoxia inducing factor-1α and transforming growth factor beta was down-regulated (all p < 0.01) by Allicin treatment.
These results suggested that Allicin protected the cardiomyocytes against HR damage by reducing apoptosis, inflammation and mitochondrial injury, thus providing a basis for its potential use in the treatment of myocardial IR.