基于文献及实验探讨急性心肌缺血模型大鼠海马神经元凋亡的研究

Study of apoptosis of hippocampal neurons in acute myocardial ischemia model rats based on literature and experiments

目的 通过文献整理分析急性心肌缺血(acute myocardial ischemia, AMI)模型建模特点及表型分析的相关指标,为本文建模提供依据;动物实验研究观察AMI大鼠海马CA1区氧化应激因子及凋亡蛋白的变化,探讨心脑同病的可能机制。方法 文献研究部分以2001年1月~2021年12月为时间范围,“Acute myocardial ischemia”“急性心肌缺血”“动物实验”为主题词,检索Web of Science、中国知网、万方、维普数据库,归纳总结AMI模型建模及表型指标等特点;动物实验部分,40只健康5~6周龄SPF级SD大鼠,雄雌各半,随机分为4个组:假手术雄鼠、假手术雌鼠、LAD雄鼠、LAD雌鼠,每组10只。LAD组大鼠进行左冠状动脉前降支(left anterior descending coronary artery, LAD)结扎,假手术组只穿线不结扎。Powerlab监测心电图(ECG)变化;HE染色观察心肌组织病理改变;ELISA法检测血清心肌酶(CK-MB、cTnI)、海马CA1区氧化应激因子(MDA、SOD、Uch-L1)水平;Western Blot法检测海马CA1区细胞凋亡蛋白(Caspase-3、PARP1及Cleaved Caspase-3、Cleaved PARP1)表达。结果 (1)411篇文献分析发现,目前研究多选择SD和Wistar大鼠进行冠状动脉前降支(LAD)结扎术制备AMI模型,表型分析指标多集中于心脏形态与功能观察,有关心脏与大脑的crosstalk研究偏少;(2)动物实验结果发现,与假手术组相比,LAD雄雌大鼠心电图ST段均明显抬高,心肌细胞水肿,心肌纤维排序紊乱,广泛炎症细胞浸润,血清CK-MB、cTnI、Uch-L1表达显著增加。海马MDA、Cleaved Caspase-3/Caspase-3及Cleaved PARP1/PARP1表达升高,SOD显著降低。结论 AMI可引起海马氧化应激,诱导神经细胞凋亡,为心脑同病提供实验依据。

Objective To provide a basis for the model in this paper,we analyzed the characteristics of acute myocardial ischemia(AMI)models and relevant phenotypic indexes in the literature.Methods In the literature research,the time range was January 2001 to December 2021;“acute myocardial ischemia”and“animal experiments”were used as the subject search terms,and Web of Science,China Knowledge Network,Wanfang,and Weipu databases were searched to summarize the characteristics of AMI modeling and phenotypic indicators.In the animal experiments,40 healthy 5⁃to 6⁃week⁃old SPF SD rats,both male and female,were randomly divided into four groups(sham male,sham female,coronary artery ligation(LAD)male,and LAD female),with 10 rats in each group.LAD group underwent ligation of the left anterior descending coronary artery(LAD),Sham group underwent the same operation,but only threading without ligation.Powerlab was used to monitor electrocardiogram(ECG)changes,and HE staining was used to observe myocardial pathological changes.ELISA was used to detect serum cardiac enzymes(CK⁃MB,cTnI)and oxidative stress factors(MDA,SOD,Uch⁃L1)in the hippocampal CA1 region.The levels of apoptotic proteins(Caspase⁃3,PARP1,Cleaved Caspase⁃3,Cleaved PARP1)in the hippocampal CA1 region were detected by Western Blot.Results An analysis of 411 papers found that most studies selected SD or Wistar rats to prepare LAD models and concentrated on morphological and functional changes to the heart,while studies on heart⁃brain crosstalk were rare.Compared with the sham⁃operation groups,LAD male and female rats showed a significantly elevated ST segment on ECG,cardiac myocyte edema,disturbed myocardial fiber sorting,extensive inflammatory cell infiltration,and expression of serum CK⁃MB,cTnI,and Uch⁃L1.The expression of MDA,Cleaved Caspase⁃3/Caspase⁃3,and cleaved PARP1/PARP1 was increased and SOD was significantly decreased in the CA1 region of the rat hippocampus.Conclusions Acute myocardial ischemia can cause hippocampal oxidative stress and induce neuronal apoptosis,and cardiovascular disease is potentially associated with brain dysfunction.