3S,3’S构型虾青素改善小鼠心肌缺血再灌注损伤的作用研究

Effect of 3S,3’S-astaxanthin on myocardial ischemia-reperfusion injury in mice

中文关键词:  3S,3’S构型虾青素  心肌缺血再灌注损伤  心肌重构  氧化应激  凋亡  炎症

英文关键词:3S,3’S-astaxanthin  myocardial ischemia-reperfusion injury  myocardial remodeling  oxidative stress  apoptosis  inflammation

中文摘要:

      目的 探讨3S,3’S构型虾青素(3S,3’S-astaxanthin, 3S,3’S-AST)对心肌缺血再灌注损伤的保护作用及其机制。方法 取SPF级ICR雄性小鼠40只, 随机分为4组, 每组10只, 分别为假手术组(Sham)、心肌缺血再灌注组(reperfusion injury, I/R)、心肌缺血再灌注给予虾青素组(I/R+AST)、虾青素组(AST)。小鼠心脏经冠状动脉左前降支结扎30 min后, 再灌注7 d。qRT-PCR检测心肌重构相关因子TGF-β1、Col Ⅰ (Collegen Ⅰ)和Col Ⅲ (Collegen Ⅲ)的mRNA表达水平, 蛋白质印迹法(western-blot)检测心肌重构、凋亡、NKA/Src/Erk1/2/ROS信号扩增环路相关蛋白Col Ⅰ、Col Ⅲ、Bcl-2、Bax、p-Src/c-Src和p-Erk1/2/Erk1/2的表达水平, 通过试剂盒检测小鼠血清中心肌损伤及氧化应激标志物乳酸脱氢酶(lactate dehydrogenase, LDH)、丙二醛(malondialdehyde, MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-PX)和肌酸激酶同工酶(creatinine kinase, CK-MB)酶活性, 通过ELISA试剂盒检测心肌组织中炎症因子IL-6、IL-1β及TNF-α的水平。结果 心肌缺血再灌注小鼠中, 心脏组织胶原含量和心肌细胞凋亡损伤明显增加, 发生心脏重构。而经口给予3S,3’S-AST能显著减低TGF-β1、Col Ⅰ和Col Ⅲ mRNA水平(P＜0.05, P＜0.001), 降低Col Ⅰ、Col Ⅲ、Bax蛋白表达并促进Bcl-2表达增高(P＜0.05, P＜0.001), 抑制Src和Erk1/2活化(P＜0.05), 减少心肌纤维化和心肌细胞凋亡。结论 3S,3’S-AST通过抑制NKA/Src/Erk1/2/ROS扩增环路, 产生抗氧化及抗炎作用, 从而减轻细胞氧化应激损伤导致的细胞凋亡和心肌纤维化, 改善I/R心肌重构。

英文摘要:

      Objective To investigate the protective effect of 3S,3’S-astaxanthin on myocardial ischemia-reperfusion injury and its mechanism. Methods Forty SPF ICR male mice were randomly divided into 4 groups (10 in each group): Sham group (Sham), I/R group (I/R), I/R group with astaxanthin (I/R+AST) and astaxanthin group (AST). The mouse heart was ligated by left anterior descending coronary artery 30 min, then gave reperfusion for 7 days. The mRNA expression levels of TGF-β1, Col I (Collegen I) and Col III (Collegen III) were detected by qRT-PCR, the levels of myocardial remodeling, apoptosis, NKA/Src/Erk1/2/ROS signal amplification loop-related proteins Col I, Col III, Bcl-2, Bax, p-Src/c-Src and p-Erk1/2/Erk1/2 were detected by western-blot, the markers of oxidative stress lactate dehydrogenase (LDH), malondialdehyde (MDA), glutathione peroxidase (GSH-PX) and creatinine kinase (CK-MB) were detected by corresponding kit, and the levels of inflammatory cytokines in myocardium IL-6, IL-1β and TNF-α were detected by ELISA kit. Results In myocardial ischemia reperfusion mice, the collagen content and apoptosis of myocardial cells were significantly increased, and cardiac remodeling occurred. Oral administration of AST could significantly reduce the TGF-β1, Col Ⅰ, and Col Ⅲ mRNA expression (P<0.05, P<0.001), reduce the Col Ⅰ, Col Ⅲ, and Bax proteins expression and promote Bcl-2 protein expression (P<0.05, P<0.001), inhibit Src and Erk1/2 activation (P<0.05), reduce myocardial fibrosis and myocardial cell apoptosis. Conclusion 3S,3’S-AST can inhibit the NKA/Src/Erk1/2/ROS amplification circuit, exert anti-oxidative and anti-inflammatory effects, reduce cell apoptosis and myocardial fibrosis induced by oxidative stress injury, and improve I/R myocardial remodeling.