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使用轻度低温对大鼠心肌梗死期间缺血和/或再灌注损伤的减轻作用:Bcl-2、Bax、Bak和TUNEL的免疫组织化学研究

Attenuation of ischemia and/or reperfusion injury during myocardial infarction using mild hypothermia in rats: an immunohistochemical study of Bcl-2, Bax, Bak and TUNEL.

作者信息

Babu Phanithi Prakash, Suzuki Gen, Ono Yukihiko, Yoshida Yasuji

机构信息

Department of Pathology, University of Hyderabad, Hyderabad 500 046, India.

出版信息

Pathol Int. 2004 Dec;54(12):896-903. doi: 10.1111/j.1440-1827.2004.01767.x.

Abstract

The aim of the present study was to determine the beneficial effect of mild hypothermia during ischemia and/or reperfusion injury in myocardial infarction. Sprague-Dawley rats (400 +/- 20 g) were subjected to 30 min occlusion of the left coronary artery followed by 24 h reperfusion. Rats were divided into normothermic (NT; 37 degrees C) and hypothermic (HT; 34 degrees C) groups. In the HT group hypothermia was maintained during coronary occlusion and continued for 30 min following reperfusion. Histological analysis revealed dead cardiomyocytes and polymorphonuclear neutrophil infiltration after 24 h. Myocardial infarction, measured using an image analyzer, showed that the percentage area of infarction was significantly decreased in the HT group. Immunohistochemical analysis was carried out using antibodies against Bcl-2, Bax and Bak. DNA fragments were labeled in situ using the 3'-OH end-labeling method (TUNEL). In the HT group Bcl-2 was induced in many myocytes, whereas Bax and Bak were induced in only a few myocytes. A higher number of TUNEL-positive cells were recorded in the NT group than in the HT group, but these were more thinly scattered in the HT group. The expression pattern revealed that many myocytes could survive at the border zone in the HT group; in contrast, few myocytes in the NT group were able to survive. Our results suggest that mild hypothermia selectively interferes with, and mitigates, reperfusion injury.

摘要

本研究的目的是确定轻度低温在心肌梗死的缺血和/或再灌注损伤过程中的有益作用。将体重400±20克的Sprague-Dawley大鼠进行左冠状动脉闭塞30分钟,随后再灌注24小时。大鼠被分为正常体温组(NT;37℃)和低温组(HT;34℃)。在HT组中,低温在冠状动脉闭塞期间维持,并在再灌注后持续30分钟。组织学分析显示24小时后出现死亡的心肌细胞和多形核中性粒细胞浸润。使用图像分析仪测量心肌梗死情况,结果显示HT组梗死面积百分比显著降低。使用抗Bcl-2、Bax和Bak的抗体进行免疫组织化学分析。采用3'-OH末端标记法(TUNEL)对DNA片段进行原位标记。在HT组中,许多心肌细胞诱导表达Bcl-2,而只有少数心肌细胞诱导表达Bax和Bak。NT组中TUNEL阳性细胞数量高于HT组,但在HT组中分布更为稀疏。表达模式显示,HT组中许多心肌细胞能够在边界区存活;相比之下,NT组中能够存活的心肌细胞很少。我们的结果表明,轻度低温可选择性地干预并减轻再灌注损伤。

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