Histochemical assessment of early myocardial infarction using 2,3,5-triphenyltetrazolium chloride in blood-perfused porcine hearts
Histochemical assessment of early myocardial infarction using 2,3,5-triphenyltetrazolium chloride in blood-perfused porcine hearts
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DOI：
10.1016/j.vascn.2006.02.010
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发表时间：
2006-11-01
影响因子：
1.9
通讯作者：
Fritz, Hans
中科院分区：
文献类型：
--
作者：
Khalil, Philipe N.;Siebeck, Matthias;Fritz, Hans
简介：尽管之前已经描述了用于肉眼检测心肌梗死的三苯基四唑（TTC）反应的化学机制，但缺乏关于正确组织制备和在完整大型动物中使用该技术的文献报告。研究方法：我们研究了血液灌注猪心TTC染色的特殊要求，验证了各种处理步骤，并为这种组织化学方法的精确和简便使用提供了详细的信息。左前降支冠状动脉闭塞45分钟，然后再灌注6小时，在开胸准备使用麻醉家猪。切除心脏，并验证器官处理步骤和TTC染色程序。结果如下：该方案包括（i）冠状动脉内盐水灌注，（ii）通过Evans蓝染料对非缺血区域进行压力控制测定，（iii）冻融循环，（iv）三苯基四唑孵育期，以及（v）使用4%多聚甲醛的漂白循环。TTC染色结果由梗死区、危险区和非危险区的过渡区的组织学证实。讨论内容：如果仔细考虑与血液灌注猪心相关的一些特殊特征，则可以获得用于随后的梗死面积计算的可靠结果，并排除大的潜在误差。(c)2006爱思唯尔公司All rights reserved.
Introduction: Although the chemical mechanism of the triphenyltetrazolium (TTC) reaction, for macroscopic detection of myocardial infarction, has been described previously, literature reports on correct tissue preparation and the use of this technique in intact large animals are lacking. Methods: We investigated the special requirements for TTC staining in blood-perfused porcine hearts, validated the various handling steps and provided detailed information for precise and easy use of this histochemical method. The left anterior descending coronary artery was occluded for 45 min followed by 6 h of reperfusion in an open chest preparation using anesthetised domestic pigs. The hearts were excised and the organ-handling steps and TTC-staining procedure validated. Results: The protocol includes (i) intracoronary saline perfusion, (ii) pressure-controlled determination of the non-ischemic region by Evans blue dye, (iii) a freeze-thaw cycle, (iv) a triphenyltetrazolium incubation period, and (v) a bleach cycle with 4% paraformaldehyde. The TTC-staining results were confirmed by histology of transitional regions of the infarct area, area-at-risk and non-risk-region. Discussion: If some special features associated with blood-perfused porcine hearts are considered carefully, reliable results for subsequent infarct size calculations can be obtained and large potential errors excluded. (c) 2006 Elsevier Inc. All rights reserved.