Methods for histological characterization of cryo-induced myocardial infarction in a rat model.
Methods for histological characterization of cryo-induced myocardial infarction in a rat model.
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DOI：
10.1016/j.acthis.2020.151624
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发表时间：
2020-10
影响因子：
2.5
通讯作者：
Joddar B
中科院分区：
文献类型：
--
作者：
Alonzo M;Delgado M;Cleetus C;Kumar SA;Thakur V;Chattopadhyay M;Joddar B
结扎左前降支（LAD）冠状动脉通常用于诱导动物心肌梗死（MI）;然而，已知结扎会导致心律失常和不可预测的坏死损伤区域。冷冻梗死是一种替代方法，已被采用来创建心肌损伤的可再现模型。在本研究中，对Sprague-Dawley大鼠进行开胸手术，然后冷冻诱导心脏梗死，而对照-假手术组仅进行开胸手术，然后从所有动物中收集心脏。组织切片用苏木精和伊红染色并分析以确定心肌密度、纤维长度和纤维曲率。观察结果显示，在梗死组织切片中，肌肉密度、心脏纤维长度和纤维扭曲减少。对组织切片进行Gomori三色染色以研究MI后对胶原蛋白的影响，其显示与假手术模型相比，实验模型的胶原蛋白染色强度增强，表明纤维化，这是心肌损伤的既定结果。用DAPI和连接蛋白-43（Cx-43）对组织切片进行免疫组织化学染色显示，与假手术样品相比，在实验样品中存在减少的DAPI染色和不太明显的Cx-43表达。结果表明，冷冻梗死导致显著的细胞损伤，随后破坏和解聚心肌细胞中的功能性Cx-43连接，这对于正常和健康的心脏生理学和功能至关重要。在大鼠模型中对冷冻诱导的MI进行的定量组织学研究可以帮助其他人尝试通过冷冻损伤优化大鼠MI模型，研究心脏疾病进展，并帮助构建工程化心脏组织。
Ligation of the left anterior descending (LAD) coronary artery has been commonly employed to induce myocardial infarction (MI) in animals; however, it is known to pose setbacks in the form of cardiac arrhythmias and unpredictable areas of necrotic damage. Cryo-infarction is an alternate method that has been adopted to create a reproducible model of a myocardial injury. In this study, Sprague-Dawley rats were subjected to thoracotomy followed by cryo-induced infarction of the heart, while the control-sham group was only subjected to thoracotomy following which the heart was collected from all animals. Tissue sections were stained with hematoxylin and eosin and analyzed to determine cardiac muscle density, fiber length, and fiber curvature. Observations revealed reduced muscle density, cardiac fiber length, and distorted fibers in infarcted tissue sections. Gomori’s Trichrome staining was performed on tissue sections to study the effects of post MI on collagen, which showed enhanced intensity of collagen staining indicating fibrosis for the experimental models as compared to the sham models, an established consequence to myocardial injury. Immunohistochemical staining of the tissue sections with DAPI and connexin-43 (Cx-43) revealed that there was reduced DAPI staining and a less pronounced expression of Cx-43 in the experimental samples as compared to the sham samples. Results implied significant cell damage resulting from the cryo-infarction, subsequently disrupting and disaggregating the functional Cx-43 junction in cardiac myocytes, which is essential for normal and healthy cardiac physiology and function. This quantitative histological study of cryo-induced MI in a rat model can aid others attempting to optimize MI models in rats via cryo-injury, to study cardiac disease progression, and to aid in the construction of engineered cardiac tissues.
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