本文關(guān)鍵詞： 脂肪間充質(zhì)干細(xì)胞 bFGF 心肌梗死 細(xì)胞移植　出處：《新鄉(xiāng)醫(yī)學(xué)院》2013年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：背景 干細(xì)胞移植是治療心肌梗死的一種非常有前景的新方法。在特定的微環(huán)境下移植間充質(zhì)于細(xì)胞可分化為血管內(nèi)皮細(xì)胞或者心肌細(xì)胞,并分泌血管生長(zhǎng)因子,促進(jìn)心肌血管生成,改善心臟功能。然而,急性心肌梗死經(jīng)干細(xì)胞移植治療后,移植干細(xì)胞在梗死區(qū)會(huì)因?yàn)槿毖?再灌注損傷、炎性因子等惡劣環(huán)境的作用而大量死亡,將極大的影響干細(xì)胞移植的療效。因此,人們?cè)噲D通過(guò)采取干預(yù)措施即利用細(xì)胞因子、藥物、化合物等對(duì)移植干細(xì)胞進(jìn)行體外預(yù)處理,或采用有利于干細(xì)胞存活、黏附或促進(jìn)血管形成的基因?qū)Ω杉?xì)胞進(jìn)行修飾,直接和間接地提高干細(xì)胞在缺血心肌微環(huán)境中的存活和分化能力,從而有助于促進(jìn)移植干細(xì)胞修復(fù)壞死心肌和改善心功能。 目的 研究經(jīng)去分化預(yù)處理誘導(dǎo)后的脂肪間充質(zhì)干細(xì)胞應(yīng)用于細(xì)胞移植治療是否能有效地提高干細(xì)胞在缺血心肌微環(huán)境中的生存率,為今后研究脂肪源性干細(xì)胞治療急性心肌梗死提供實(shí)驗(yàn)依據(jù)。 方法 1分離雌性小鼠(30-50g體重)腹股溝皮下脂肪組織用于脂肪問(wèn)充質(zhì)干細(xì)胞分離與培養(yǎng)；取第3代脂肪間充質(zhì)干細(xì)胞用于本實(shí)驗(yàn)。 2結(jié)扎雌性SD大鼠的左冠狀動(dòng)脈前降支,建立急性心肌梗死模型；在心肌梗死模型建立后1小時(shí),于梗死區(qū)邊緣分別移植單純ADMSCs、5-aza預(yù)處理的ADMSCs和堿性成纖維細(xì)胞生長(zhǎng)因子(bFGF)預(yù)處理的ADMSCs;根據(jù)細(xì)胞預(yù)處理方式不同隨機(jī)分組進(jìn)行實(shí)驗(yàn)觀察；在細(xì)胞移植后1、2、3和4周,用M型超聲心動(dòng)圖檢測(cè)心功能變化。在動(dòng)脈結(jié)扎后不同時(shí)間取梗死區(qū)心肌組織,作HE染色和Masson染色,觀察梗死區(qū)心肌組織的病理學(xué)變化；熒光顯微鏡下驗(yàn)證移植后的ADMSCs存活情況。 結(jié)果 1體外培養(yǎng)的脂肪源性干細(xì)胞表達(dá)CD29、CD90,不表達(dá)CD45；證明實(shí)驗(yàn)所使用細(xì)胞為脂肪間充質(zhì)干細(xì)胞。 2心肌梗死模型：冠狀動(dòng)脈前降支結(jié)扎后,左室前壁變蒼白,心臟膨大,左心耳充血,心室收縮減弱,與未梗死區(qū)界線清楚,表明心肌梗死動(dòng)物模型制備成功。 3移植細(xì)胞存活情況和心功能觀察：心功能檢測(cè)發(fā)現(xiàn),在ADMSCs移植后1周,與模型對(duì)照組相比,bFGF預(yù)處理ADMSCs移植組左室舒張末期內(nèi)徑(LVDd)左室收縮期內(nèi)徑(LVDs)無(wú)明顯縮小,左室射血分?jǐn)?shù)(LVEF)及左室短軸縮短率(LVFS)也無(wú)明顯提高,兩組無(wú)顯著性差異(n=10,p0.05)。而在細(xì)胞移植后4周,bFGF預(yù)處理ADMSCs移植組LVDd和LVDs較1周移植組和模型對(duì)照組組均有明顯縮小,EF及FS也明顯提高,與一周移植組和模型對(duì)照組以及單純ADMSCs移植組相比具有顯著性差異(n=10, p0.05)。HE染色可看到心肌梗死區(qū)域瘢痕增生紊亂,新生血管形成。 結(jié)論 經(jīng)去分化預(yù)處理誘導(dǎo)后的脂肪干細(xì)胞移植治療心肌梗死,有利于移植細(xì)胞在受損心肌內(nèi)的存活,并促進(jìn)心功能恢復(fù),其療效優(yōu)于單純干細(xì)胞移植。
[Abstract]:Background. Stem cell transplantation is a promising new way to treat myocardial infarction. Transplantation of mesenchymal cells in specific microenvironments can differentiate into vascular endothelial cells or cardiomyocytes and secrete vascular growth factors. Promote myocardial angiogenesis and improve cardiac function. However, after transplantation of stem cells in acute myocardial infarction, stem cells in the infarct area will die in large numbers because of the effects of ischemia / reperfusion injury, inflammatory factors and other adverse circumstances. It's going to have a huge impact on the effectiveness of stem cell transplantation. So people are trying to pretreat the transplanted stem cells in vitro by using cytokines, drugs, compounds and so on, or by using them that are good for the survival of stem cells. The adhesion or promotion of angiogenic genes modifies the stem cells, directly and indirectly improves the viability and differentiation of stem cells in the ischemic myocardial microenvironment, thus facilitating the transplantation of stem cells to repair the necrotic myocardium and improve the cardiac function. Purpose. To study whether the application of adipose mesenchymal stem cells induced by dedifferentiation pretreatment in cell transplantation can effectively improve the survival rate of stem cells in ischemic myocardial microenvironment. To provide experimental evidence for the treatment of acute myocardial infarction by adipose-derived stem cells. Method. 1 the inguinal subcutaneous adipose tissue was used for the isolation and culture of adipose mesenchymal stem cells, and the third generation adipose mesenchymal stem cells were used in this experiment. 2ligation of left anterior descending coronary artery of female SD rats to establish acute myocardial infarction model, 1 hour after myocardial infarction model was established, ADMSCs pretreated by ADMSCs5-aza and ADMSCs pretreated by basic fibroblast growth factor (bFGF) were transplanted at the edge of infarct area respectively. Cardiac function was detected by M-mode echocardiography. Myocardial tissue of infarcted area was taken at different time after artery ligation and stained with HE and Masson to observe the pathological changes of myocardial tissue in infarcted area. The survival of ADMSCs after transplantation was verified by fluorescence microscope. Results. 1 the adipose-derived stem cells expressed CD29 CD90, but not CD45, which proved that the cells used in the experiment were adipose mesenchymal stem cells. 2Myocardial infarction model: after ligation of anterior descending coronary artery, left ventricular anterior wall became pale, heart dilated, left atrial appendage congested, ventricular contraction weakened, and the boundary between left atrial appendage and uninfarcted area was clear, which indicated that myocardial infarction animal model was successfully prepared. 3The survival of transplanted cells and cardiac function: at 1 week after ADMSCs transplantation, the left ventricular end-diastolic diameter (LVDD) and left ventricular systolic diameter (LVDss) were not significantly decreased in the ADMSCs group pretreated with bFGF compared with the model control group. Left ventricular ejection fraction (LVEF) and left ventricular shortening rate (LVFS) were not significantly increased. There was no significant difference between the two groups, but the LVDd and LVDs in the ADMSCs transplantation group pretreated with bFGF 4 weeks after transplantation were significantly smaller than those in the 1-week transplantation group and the model control group. Compared with the one-week transplantation group, the model control group and the simple ADMSCs group, there was a significant difference between the two groups. The hypertrophic scar and neovascularization in the myocardial infarction area could be seen by HE staining. Conclusion. The transplantation of adipose stem cells induced by dedifferentiation preconditioning is beneficial to the survival of transplanted cells in the injured myocardium and to the recovery of cardiac function, and the effect is better than that of stem cell transplantation alone.
【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】：2013
【分類(lèi)號(hào)】：R542.22

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