Valentin Jérémy, Frobert Aurélien, Ajalbert Guillaume, Cook Stéphane, Giraud Marie-Noëlle
Cardiology, Department of Medicine, University of Fribourg.
Cardiology, Department of Medicine, University of Fribourg;
J Vis Exp. 2016 Dec 11(118):54914. doi: 10.3791/54914.
Myocardial infarction is defined as cardiomyocyte death due to prolonged ischemia; an inflammatory response and scar formation (fibrosis) follow the ischemic injury. Following the initial acute phase, chronic remodeling of the left ventricle (LV) modifies the structure and function of the heart. Permanent coronary ligation in small animals has been widely used as a reference model for a chronic model of MI. Thinning of the infarcted wall progressively develops to transmural fibrosis. Histological assessment of infarct size is commonly performed; nevertheless, a standardization of the methods for quantification is missing. Indeed, important methodological aspects, such as the number of sections analyzed and the sampling and quantification methods, are usually not described and therefore preclude comparison across investigations. Too often, quantification is performed on a single section obtained at the level of the papillary muscles. Because novel strategies aimed at reducing infarct expansion and remodeling are under investigation, there is an important need for the standardization of accurate heart sampling protocols. We describe an accurate method to quantify the infarct size using a systematic sampling of harvested rat heart and image analyses of trichromatic stained histological sections obtained from base to apex. We also provide evidence that calculating the expansion index (EI) allowed for infarct size assessment, taking into account changes of the left ventricle throughout the remodeling.
心肌梗死定义为由于长时间缺血导致的心肌细胞死亡;缺血性损伤后会出现炎症反应和瘢痕形成(纤维化)。在初始急性期之后,左心室(LV)的慢性重塑会改变心脏的结构和功能。小动物永久性冠状动脉结扎已被广泛用作心肌梗死慢性模型的参考模型。梗死壁变薄逐渐发展为透壁纤维化。通常会对梗死面积进行组织学评估;然而,量化方法缺乏标准化。实际上,重要的方法学方面,如分析的切片数量以及取样和量化方法,通常未被描述,因此无法在不同研究之间进行比较。很多时候,量化是在乳头肌水平获得的单个切片上进行的。由于旨在减少梗死扩展和重塑的新策略正在研究中,因此迫切需要标准化准确的心脏取样方案。我们描述了一种准确的方法,通过对收获的大鼠心脏进行系统取样以及对从心底到心尖的三色染色组织学切片进行图像分析来量化梗死面积。我们还提供了证据,即计算扩展指数(EI)可以在考虑左心室在整个重塑过程中的变化的情况下进行梗死面积评估。