【摘要】：背景近年來,干細胞移植治療心肌梗死成為一大研究熱點。動員心肌內自身干細胞再生和修復成為了目前爭論和研究的焦點,同時也決定了未來干細胞治療心臟病方法的多樣性。目前關于心肌干細胞研究較多的有sca-1陽性心肌干細胞,c-kit陽性心肌干細胞以及nanog陽性心肌干細胞。它們具有分化成心肌細胞的潛能。但心肌梗死后心肌干細胞的動態(tài)變化如何?它們在干細胞標記表達方面是否存在共表達情況?這些問題均有待進一步研究探討。目的探討大鼠急性心肌梗死后不同時段心肌梗死區(qū)域心肌干細胞的動態(tài)變化及其標記物共表達情況。方法健康成年SD大鼠25只,隨機分為正常對照組(n=5)和急性心肌梗死組(n=20)。結扎大鼠冠狀動脈前降支制備急性心肌梗死模型,術前和術后1w、2w、3w和4w分別檢測左室射血分數(shù)(EF)、左室短軸縮短率(FS)、左室舒張末期內徑(LVID;d)、左室舒張末期容積(LV Vol;d)和左室舒張末期后壁厚度(LVPW;d)。利用免疫組化技術對各組心臟切片進行免疫顯色,觀察sca-1陽性心肌干細胞的動態(tài)變化并進行計數(shù)分析。利用免疫熒光雙標技術觀察sca-1、c-kit、nanog的共表達情況。運用Western Blotting技術檢測c-kit、nanog、sca-1蛋白的表達水平。結果1心肌梗死模型組EF、FS、LVPW;d均下降(P0.05)。LVID;d、LV Vol;d逐漸上升(P0.05)。2免疫組化結果顯示nanog、c-kit、sca-1陽性心肌干細胞數(shù)量在2w時上升至高峰,隨后下降。在梗死早期陽性細胞散在表達,3w時在梗死區(qū)聚集成團或條帶狀。3免疫熒光結果顯示,梗死區(qū)域nanog和c-kit蛋白、sca-1和c-kit蛋白、sca-1和nanog蛋白之間在部分細胞上存在共表達。4 Western Blotting結果顯示c-kit、nanog、sca-1蛋白含量于2w時達高峰,與免疫組化結果基本一致。結論1心肌干細胞隨梗死時程的變化而變化,干細胞有向梗死區(qū)域遷移和聚集的傾向。2一種干細胞可表達兩種干細胞的表面標記,提示心肌干細胞的亞群可能存在交叉重疊,其功能可能具有多樣性。
[Abstract]:Background in recent years, stem cell transplantation has become a hot topic in the treatment of myocardial infarction. Mobilization of myocardial stem cells regeneration and repair has become the focus of debate and research, but also determine the future diversity of stem cells in the treatment of heart disease. At present, there are more sca-1 positive myocardial stem cells, c kit positive myocardial stem cells and nanog positive myocardial stem cells. They have the potential to differentiate into cardiomyocytes. But what about the dynamic changes of myocardial stem cells after myocardial infarction? Do they co-express stem cell markers? These problems need to be further studied and discussed. Objective to investigate the dynamic changes of myocardial stem cells and co-expression of markers in myocardial infarction region after acute myocardial infarction in rats. Methods 25 healthy adult SD rats were randomly divided into two groups: normal control group (n = 5) and acute myocardial infarction group (n = 20). Acute myocardial infarction model was established by ligating anterior descending coronary artery in rats. Left ventricular ejection fraction (EF), left ventricular short-axis shortening rate (FS), left ventricular end-diastolic volume (LV Vol;d), left ventricular end-diastolic volume (LV Vol;d) and left ventricular posterior wall thickness (LVPW;d) were measured before and 1 week after operation. Immunohistochemical technique was used to detect the dynamic changes of sca-1 positive myocardial stem cells in each group. The co-expression of sca-1,c-kit,nanog was observed by immunofluorescence double labeling technique. Western Blotting technique was used to detect the expression level of c-kitoku nanogCa-1 protein. Results 1in myocardial infarction model group, EF,FS,LVPW;d decreased (P0.05). LV Vol;d increased gradually (P0.05). 2 the results of immunohistochemistry showed that the number of nanog,c-kit,sca-1 positive myocardial stem cells increased to the peak at 2 weeks and then decreased. The results of immunofluorescence showed that the positive cells were clustered or banded in the infarct area at 3w after the positive cells were expressed in the early stage of infarction. There was coexpression of 4. 4 Western Blotting in some cells between nanog and c-kit protein, sca-1 and nanog protein in the infarcted area. The results showed that the content of c-kitacia nanogogsca-1 protein reached its peak at 2 weeks, which was consistent with the immunohistochemical results. Conclusion 1 Myocardial stem cells change with the change of infarct duration. 2. One kind of stem cells can express the surface markers of two kinds of stem cells, suggesting that the subsets of myocardial stem cells may have overlapping. Its functions may be diverse.
【學位授予單位】：新鄉(xiāng)醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R542.22

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