本文選題：骨髓間充質(zhì)細胞 + 細胞分化�。� 參考：《大連醫(yī)科大學》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。雖然成熟心肌細胞不是終末分化細胞，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)，進而促進心力衰竭發(fā)生，最終造成患者死亡。現(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(PTCA)、冠狀動脈旁路移植術(CABG)及心臟移植術等。前3項方法可改善心肌供血，但不能使壞死心肌復生，而心臟移植雖可替代受損心臟，但難以廣泛的推廣應用。近年來，隨著分子生物學的發(fā)展，利用成體干細胞種植方法來增加梗死部位心肌細胞的數(shù)量，從而改善心功能及降低AMI后遠期病死率，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胞來源主要有骨髓干細胞、骨骼肌衛(wèi)星細胞、平滑肌干細胞、胚胎干細胞以及胎兒心肌細胞。骨髓干細胞在人體廣泛分布而且容易獲取，種植中無免疫排斥反應，也不存在倫理學的爭論，因此它們在細胞性心肌整復的臨床應用中很有優(yōu)勢。本研究探討骨髓源性的成體干細胞體外在5-氮胞苷、中藥三七皂甙的誘導下轉(zhuǎn)化為心肌樣細胞的可行性，并將骨髓干細胞與心肌細胞共培養(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)干細胞(marrow mesenchymal stem cells，MSCs)體外誘導分化為心肌樣細胞 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，觀察細胞形態(tài)學變化：在培養(yǎng)3周時，用免疫細胞化學染色的方法檢測細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)進行鑒定；及電鏡下觀察細胞內(nèi)部結(jié)構(gòu)。取SD大鼠心肌細胞作為陽性對照，，未經(jīng)5-氮胞苷誘導正常培養(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.
【學位授予單位】：大連醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2006
【分類號】：R542.22;R329

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