【摘要】：背景近年來(lái),干細(xì)胞移植治療心肌梗死成為一大研究熱點(diǎn)。動(dòng)員心肌內(nèi)自身干細(xì)胞再生和修復(fù)成為了目前爭(zhēng)論和研究的焦點(diǎn),同時(shí)也決定了未來(lái)干細(xì)胞治療心臟病方法的多樣性。目前關(guān)于心肌干細(xì)胞研究較多的有sca-1陽(yáng)性心肌干細(xì)胞,c-kit陽(yáng)性心肌干細(xì)胞以及nanog陽(yáng)性心肌干細(xì)胞。它們具有分化成心肌細(xì)胞的潛能。但心肌梗死后心肌干細(xì)胞的動(dòng)態(tài)變化如何?它們?cè)诟杉?xì)胞標(biāo)記表達(dá)方面是否存在共表達(dá)情況?這些問(wèn)題均有待進(jìn)一步研究探討。目的探討大鼠急性心肌梗死后不同時(shí)段心肌梗死區(qū)域心肌干細(xì)胞的動(dòng)態(tài)變化及其標(biāo)記物共表達(dá)情況。方法健康成年SD大鼠25只,隨機(jī)分為正常對(duì)照組(n=5)和急性心肌梗死組(n=20)。結(jié)扎大鼠冠狀動(dòng)脈前降支制備急性心肌梗死模型,術(shù)前和術(shù)后1w、2w、3w和4w分別檢測(cè)左室射血分?jǐn)?shù)(EF)、左室短軸縮短率(FS)、左室舒張末期內(nèi)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(shù)對(duì)各組心臟切片進(jìn)行免疫顯色,觀(guān)察sca-1陽(yáng)性心肌干細(xì)胞的動(dòng)態(tài)變化并進(jìn)行計(jì)數(shù)分析。利用免疫熒光雙標(biāo)技術(shù)觀(guān)察sca-1、c-kit、nanog的共表達(dá)情況。運(yùn)用Western Blotting技術(shù)檢測(cè)c-kit、nanog、sca-1蛋白的表達(dá)水平。結(jié)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(jié)果顯示nanog、c-kit、sca-1陽(yáng)性心肌干細(xì)胞數(shù)量在2w時(shí)上升至高峰,隨后下降。在梗死早期陽(yáng)性細(xì)胞散在表達(dá),3w時(shí)在梗死區(qū)聚集成團(tuán)或條帶狀。3免疫熒光結(jié)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(xì)胞上存在共表達(dá)。4 Western Blotting結(jié)果顯示c-kit、nanog、sca-1蛋白含量于2w時(shí)達(dá)高峰,與免疫組化結(jié)果基本一致。結(jié)論1心肌干細(xì)胞隨梗死時(shí)程的變化而變化,干細(xì)胞有向梗死區(qū)域遷移和聚集的傾向。2一種干細(xì)胞可表達(dá)兩種干細(xì)胞的表面標(biāo)記,提示心肌干細(xì)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(xué)位授予單位】：新鄉(xiāng)醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R542.22

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