实验性心肌缺血早期心肌的法医病理学研究

发布时间：2018-07-17 04:53

【摘要】： 目的观察大鼠急性心肌缺血模型早期正常区、交界区和缺血区心肌的病理改变，ET和NOS的表达和血浆NO浓度的变化，试图为急性心肌缺血猝死的法医病理学鉴定提供客观的形态学依据。方法 70只成年Wistar大鼠随机分为7组：0h组、0.5h组、1h组、2h组、3h组、4h组和6h组，每组10只，建立急性心肌缺血动物模型。0h组为对照组：开胸，暴露心脏，丝线绕过冠状动脉，但不结扎。其它6组为实验组：开胸，暴露心脏，结扎冠状动脉。各组在结扎冠状动脉后0.5h、1h、2h、3h、4h和6h处死。取正常区、交界区和缺血区的心肌制作标本：(1)透射电镜观察；(2)H-E染色，光镜观察；(3)ET、iNOS和eNOS免疫组化染色；(4)流式细胞仪检测细胞凋亡。分别在结扎冠状动脉前和处死大鼠时静脉取血，检测血浆中NO浓度。结果 (1)随着缺血时间的延长，大鼠血浆中NO浓度先降低，后升高，而后又降低，和对照组相比有显著性差异(p＜0.05)。(2)H-E染色未见典型的AMI组织学变化。(3)免疫组化染色发现：ET在1h组和2h组的缺血区和交界区为阳性表达，正常区和对照组为阴性表达。eNOS在正常区和对照组为阳性表达，在缺血区随缺血时间的延长表达减少。iNOS在正山西医科大学硕士学位论文2002 常区无表达，在缺血区随缺血时间的延长，表达范围和强度都越来越大。(4)缺血区和交界区的凋亡指数明显高于对照组 (p0.05)。(5)电镜观察:线粒体肿胀、空泡样变性，糖原减少和细胞核中的染色质周边化，以上变化随着缺血时间的延长逐渐加重。结论(1)H一E染色未见典型的AMI组织学变化。(2)免疫组化染色发现:ET、eNOS和1 NOS在AM工的早期有显著性变化。(3)缺血区和交界区心肌细胞的凋亡指数要高于对照组。故此，通过流式细胞仪检测心肌的凋亡指数并结合超敏SP法对心肌进行ET、iNOS和eNOS联合免疫组化染色，可以辅助诊断AMI所致的碎死。
[Abstract]:Objective to observe the changes of endothelin (et), nitric oxide synthase (NOS) and plasma no concentration in the early stage of acute myocardial ischemia in rats. This paper attempts to provide an objective morphological basis for forensic pathological identification of sudden cardiac ischemic death. Methods 70 adult Wistar rats were randomly divided into 7 groups: 0: 0h group, 0.5 h group, 1h group, 3h group, 6h group, 10 rats in each group. The animal model of acute myocardial ischemia was established as control group: open chest, expose heart, bypass coronary artery. But no ligation. The other 6 groups were treated with open chest, exposed heart and ligated coronary artery. The rats in each group were killed 0.5 h after ligation of coronary artery at 1 hour, 2 h, 3 h, 4 h and 6 h after ligation. The myocardial specimens from normal, junctional and ischemic areas were as follows: (1) transmission electron microscopy; (2) H-E staining, light microscopy; (3) ETH iNOS and Enos immunohistochemical staining; (4) flow cytometry to detect apoptosis. Venous blood was taken before ligation of coronary artery and rats were killed. No concentration in plasma was measured. Results (1) with the prolongation of ischemic time, the concentration of no in plasma of rats decreased first, then increased, then decreased. There were no typical histological changes in H-E staining compared with control group (p < 0. 05). (2). (3) Immunohistochemical staining showed that the positive expression of). (et was found in the ischemic and junctional areas of 1 h and 2 h groups. The positive expression of Enos was found in normal area and control group. The expression of iNOS decreased with the prolongation of ischemic time in ischemic area. Discussion on the Master's degree of iNOS in Shanxi Medical University No expression was found in Wen 2002, In the ischemic area, with the prolongation of the ischemic time, (4) the apoptotic index in the ischemic and junctional regions was significantly higher than that in the control group. (p0. 05). (5) Electron microscopic observation: mitochondria swelling, Vacuolar degeneration, glycogen reduction, and chromatin periphery in the nucleus, The above changes became more and more serious with the prolongation of ischemic time. Conclusion (1) there were no typical histological changes in AMI by H-E staining. (2) Immunohistochemical staining showed that there were significant changes of Enos and 1NOS in AM workers in the early stage of AM. (3) the apoptotic index of myocardial cells in ischemic and borderline areas was higher than that in control group. Therefore, the apoptosis index of myocardium was detected by flow cytometry and immunohistochemical staining of ETI iNOS and Enos was performed in combination with high sensitive SP method. It can assist in diagnosis of broken death caused by AMI.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2002
【分类号】：D919

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