【摘要】： 目的 觀察大鼠急性心肌缺血模型早期正常區(qū)、交界區(qū)和缺血區(qū)心肌的病理改變，ET和NOS的表達和血漿NO濃度的變化，試圖為急性心肌缺血猝死的法醫(yī)病理學鑒定提供客觀的形態(tài)學依據(jù)。 方法 70只成年Wistar大鼠隨機分為7組：0h組、0.5h組、1h組、2h組、3h組、4h組和6h組，每組10只，建立急性心肌缺血動物模型。0h組為對照組：開胸，暴露心臟，絲線繞過冠狀動脈，但不結扎。其它6組為實驗組：開胸，暴露心臟，結扎冠狀動脈。各組在結扎冠狀動脈后0.5h、1h、2h、3h、4h和6h處死。取正常區(qū)、交界區(qū)和缺血區(qū)的心肌制作標本：(1)透射電鏡觀察；(2)H-E染色，光鏡觀察；(3)ET、iNOS和eNOS免疫組化染色；(4)流式細胞儀檢測細胞凋亡。分別在結扎冠狀動脈前和處死大鼠時靜脈取血，檢測血漿中NO濃度。 結果 (1)隨著缺血時間的延長，大鼠血漿中NO濃度先降低，后升高，而后又降低，和對照組相比有顯著性差異(p＜0.05)。(2)H-E染色未見典型的AMI組織學變化。(3)免疫組化染色發(fā)現(xiàn)：ET在1h組和2h組的缺血區(qū)和交界區(qū)為陽性表達，正常區(qū)和對照組為陰性表達。eNOS在正常區(qū)和對照組為陽性表達，在缺血區(qū)隨缺血時間的延長表達減少。iNOS在正 山西醫(yī)科大學碩士學位論文2002 常區(qū)無表達，在缺血區(qū)隨缺血時間的延長，表達范圍和強度都 越來越大。(4)缺血區(qū)和交界區(qū)的凋亡指數(shù)明顯高于對照組 (p0.05)。(5)電鏡觀察:線粒體腫脹、空泡樣變性，糖原 減少和細胞核中的染色質周邊化，以上變化隨著缺血時間的延 長逐漸加重。 結論(1)H一E染色未見典型的AMI組織學變化。(2)免 疫組化染色發(fā)現(xiàn):ET、eNOS和1 NOS在AM工的早期有顯著性變 化。(3)缺血區(qū)和交界區(qū)心肌細胞的凋亡指數(shù)要高于對照組。 故此，通過流式細胞儀檢測心肌的凋亡指數(shù)并結合超敏SP法 對心肌進行ET、iNOS和eNOS聯(lián)合免疫組化染色，可以輔助診 斷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.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2002
【分類號】：D919

【引證文獻】

相關碩士學位論文 前1條

1 楊瑩;大鼠急性心肌缺血組織誘導型一氧化氮合酶和硝基酪氨酸的表達及其抗原穩(wěn)定性的變化[D];河北醫(yī)科大學;2010年

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