本文選題：骨髓間充質(zhì)細(xì)胞 + 細(xì)胞分化��； 參考：《大連醫(yī)科大學(xué)》2006年博士論文

【摘要】：目的：急性心肌梗死(AMI)是危害人類健康的主要疾病之一。雖然成熟心肌細(xì)胞不是終末分化細(xì)胞，但也只有有限的分裂、增殖能力。AMI患者即使有幸渡過急性期，但可由于心肌梗死(MI)后梗死部位瘢痕形成及殘留心肌代償性肥厚，引起心室重構(gòu)，進(jìn)而促進(jìn)心力衰竭發(fā)生，最終造成患者死亡�，F(xiàn)今用于AMI治療的方法通常為溶栓治療、經(jīng)皮冠狀動脈成形術(shù)(PTCA)、冠狀動脈旁路移植術(shù)(CABG)及心臟移植術(shù)等。前3項(xiàng)方法可改善心肌供血，但不能使壞死心肌復(fù)生，而心臟移植雖可替代受損心臟，但難以廣泛的推廣應(yīng)用。近年來，隨著分子生物學(xué)的發(fā)展，利用成體干細(xì)胞種植方法來增加梗死部位心肌細(xì)胞的數(shù)量，從而改善心功能及降低AMI后遠(yuǎn)期病死率，為AMI的治療開辟了新的前景。目前，用于心臟疾病方面的成體干細(xì)胞來源主要有骨髓干細(xì)胞、骨骼肌衛(wèi)星細(xì)胞、平滑肌干細(xì)胞、胚胎干細(xì)胞以及胎兒心肌細(xì)胞。骨髓干細(xì)胞在人體廣泛分布而且容易獲取，種植中無免疫排斥反應(yīng)，也不存在倫理學(xué)的爭論，因此它們在細(xì)胞性心肌整復(fù)的臨床應(yīng)用中很有優(yōu)勢。本研究探討骨髓源性的成體干細(xì)胞體外在5-氮胞苷、中藥三七皂甙的誘導(dǎo)下轉(zhuǎn)化為心肌樣細(xì)胞的可行性，并將骨髓干細(xì)胞與心肌細(xì)胞共培養(yǎng)，探討“心肌樣”微環(huán)境對這一轉(zhuǎn)化過程的影響。 方法： 一．骨髓間充質(zhì)干細(xì)胞(marrow mesenchymal stem cells，MSCs)體外誘導(dǎo)分化為心肌樣細(xì)胞 1．體外分離、培養(yǎng)SD大鼠MSCs。 2．分別用不同濃度的5-氮胞苷(5-azacytidine，5-aza)對MSCs定向誘導(dǎo)，觀察細(xì)胞形態(tài)學(xué)變化：在培養(yǎng)3周時(shí)，用免疫細(xì)胞化學(xué)染色的方法檢測細(xì)胞中的心肌特異性肌鈣蛋白T(cTnT)；用RT-PCR對心肌肌球蛋白重鏈(β-MHC)進(jìn)行鑒定；及電鏡下觀察細(xì)胞內(nèi)部結(jié)構(gòu)。取SD大鼠心肌細(xì)胞作為陽性對照，，未經(jīng)5-氮胞苷誘導(dǎo)正常培養(yǎng)的MSCs作為陰性對照。
[Abstract]:Objective: acute myocardial infarction (AMI) is one of the major diseases endangering human health. Although mature cardiomyocytes are not terminal differentiation cells, they are also limited in division. The proliferative ability of AMI patients may be due to scar formation and residual myocardial compensatory hypertrophy after myocardial infarction (MI), even though they are lucky enough to pass through the acute phase. It causes ventricular remodeling, which in turn promotes heart failure and ultimately leads to death. Nowadays, thrombolytic therapy, percutaneous transluminal coronary angioplasty (PTCA), coronary artery bypass grafting (CABG) and heart transplantation are commonly used in the treatment of AMI. The first three methods can improve the blood supply of myocardium, but can not regenerate the necrotic myocardium. Although heart transplantation can replace the damaged heart, it is difficult to be widely used. In recent years, with the development of molecular biology, the method of adult stem cell implantation is used to increase the number of myocardial cells in the infarct area, thus improve the cardiac function and reduce the long-term mortality after AMI, which opens up a new prospect for the treatment of AMI. At present, adult stem cells are mainly derived from bone marrow stem cells, skeletal muscle satellite cells, smooth muscle stem cells, embryonic stem cells and fetal cardiomyocytes. Bone marrow stem cells are widely distributed and easy to obtain in human body. There is no immune rejection in implantation and there is no ethical debate, so they have advantages in the clinical application of cellular myocardial repair. The aim of this study was to investigate the feasibility of transforming bone marrow derived adult stem cells into cardiomyocyte-like cells induced by 5-azacytidine and Panax notoginseng saponins in vitro, and co-cultured bone marrow stem cells with cardiomyocytes. To investigate the effect of myocardial microenvironment on this transformation process. Methods: I. Differentiation of bone marrow mesenchymal stem cells into cardiomyoid cells in vitro 1. MSCs of SD rats were isolated and cultured in vitro. 2. MSCs was induced by 5-azacytidine 5-azazacydine 5-aza at different concentrations, and the morphological changes were observed. The specific cardiac troponin TcTnTnTnTnTX was detected by immunocytochemical staining, the cardiac myosin heavy chain (尾 -MHCs) was identified by RT-PCR, and the structure of the cells was observed under electron microscope. SD rat cardiomyocytes were used as positive control and normal cultured MSCs without 5-azacytidine as negative control.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2006
【分類號】：R542.22;R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 姚麟,陳運(yùn)賢,彭愛華;造血干細(xì)胞移植后免疫功能重建[J];國外醫(yī)學(xué)(內(nèi)科學(xué)分冊);2000年04期

2 陳朝鳳，陳潔文，李小英;三七皂甙抗心律失常的實(shí)驗(yàn)研究[J];廣州中醫(yī)學(xué)院學(xué)報(bào);1994年02期

3 費(fèi)震宇,王文健,陳響中;三七用于冠心病治療的藥理研究[J];時(shí)珍國藥研究;1997年01期

4 鐘雪云,陳運(yùn)賢;臍帶血造血干細(xì)胞移植的倫理問題[J];中國醫(yī)學(xué)倫理學(xué);2000年05期

5 鄧漪平，羅穎祖，關(guān)超然，梁育民，張濤;LSCM觀察三七總皂甙及其組分對游離鈣離子影像變化的影響[J];中國藥理學(xué)通報(bào);1996年05期

6 羅健東,梁京生,鄭平香,聶勝利,鐘南山;三七皂甙對慢性缺氧性肺動脈高壓的影響[J];中國藥理學(xué)通報(bào);1997年04期

7 饒曼人;抗心肌缺血藥物研究的評價(jià)[J];藥學(xué)學(xué)報(bào);1985年12期

8 陳運(yùn)賢;21世紀(jì)細(xì)胞療法的展望[J];中國腫瘤;2000年02期

9 劉青，鄧漪平;三七總皂甙對血管內(nèi)皮產(chǎn)生組織型纖溶酶原激活物的影響[J];中華血液學(xué)雜志;1994年08期

10 但漢雄;三七二醇甙對大鼠實(shí)驗(yàn)性心肌梗塞的保護(hù)作用[J];中藥藥理與臨床;1993年04期

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