本文關(guān)鍵詞：低氧促進人骨髓間充質(zhì)干細胞增殖和分化及其機制研究　出處：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2005年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 低氧 增殖 分化 骨髓間充質(zhì)干細胞

【摘要】：氧是維持生命的必要條件,也是細胞功能的一種重要生理調(diào)節(jié)因子,而低氧是生命發(fā)育的基本環(huán)境。例如,在妊娠的早期,由于沒有血管的形成,所以胚胎是在低氧環(huán)境中發(fā)育的。低氧作為一種生理性的刺激因素,影響著胚胎的發(fā)生、發(fā)育及正常功能的維持。低氧可激活低氧誘導(dǎo)因子(HIF),從而調(diào)控一系列與低氧相關(guān)基因的表達。例如,誘導(dǎo)與葡萄糖分解、轉(zhuǎn)運、紅細胞和血管再生等相關(guān)的基因,使其表達增加,從而維持體內(nèi)氧環(huán)境的穩(wěn)態(tài)。關(guān)于低氧作用的研究,目前多集中在低氧對細胞的損傷和有關(guān)的適應(yīng)機制方面,而對低氧在細胞增殖、分化中影響的研究較少。特別是對于體內(nèi)胚胎發(fā)生時期和成年后的一些生理、病理過程中,出現(xiàn)細胞局部微環(huán)境低氧的情況重視不足。干細胞包括骨髓間充質(zhì)干細胞(MSCs)體外培養(yǎng)和增殖分化作用的研究都是在常氧(20%O_2)條件下進行的,這樣難以反映體內(nèi)的真實情況。低氧在體內(nèi)作為一種生理性的刺激因素,對體外培養(yǎng)的MSCs增殖和分化的作用目前還不清楚,也沒有引起科研人員的足夠重視。而人骨髓間充質(zhì)干細胞(hMSCs)是目前臨床應(yīng)用前景最好的成體干細胞,因此,本文就低氧對hMSCs體外增殖和分化作用進行了初步研究。 1.低氧對hMSCs體外增殖的作用 探討不同方式低氧對hMSCs體外增殖的影響。實驗采用血球計數(shù)板計數(shù)法和流式細胞術(shù)分別觀察了間歇性低氧(3%O_2、10%O_2)和持續(xù)性低氧(3%O_2、10%O_2、100μmol/L CoCl_2、200μmol/L CoCl_2)對hMSCs數(shù)量以及增殖指數(shù)的影響。結(jié)果發(fā)現(xiàn),間歇性低氧對hMSCs數(shù)量和增殖指數(shù)無明顯影響;持續(xù)性低氧各組hMSCs數(shù)量和增殖指數(shù)均增加(P0.05)。提示持續(xù)性低氧可促進體外培養(yǎng)的hMSCs增殖,說明持續(xù)性低氧作為體外的一種可控制因素,可調(diào)節(jié)hMSCs的體外增殖,對于hMSCs的臨床應(yīng)用具有一定的意義。 同時本研究還應(yīng)用基因芯片技術(shù)分析了低氧對hMSCs增殖過程中基因表達譜的變化。結(jié)果顯示21329個基因中282個基因有差異性表達。從基因?qū)W的角度證實了低氧對hMSCs增殖的影響是一個多基因參與的復(fù)雜過程。并從基因芯片的結(jié)果中篩選出5個目的基因,用RT-PCR方法檢測了這5個基因的表達變化。發(fā)現(xiàn)在
[Abstract]:Oxygen is a necessary condition for life and an important physiological regulator of cell function, and hypoxia is the basic environment for life development. For example, in the early stage of pregnancy, there is no blood vessel formation. Therefore, embryos are developed in hypoxic environment. As a physiological stimulant, hypoxia affects the development, development and maintenance of normal function of embryos. Hypoxia can activate hypoxia inducible factor (HIFs). It regulates the expression of a series of genes related to hypoxia, such as inducing and increasing the expression of genes related to glucose decomposition, transport, erythrocyte and vascular regeneration. So as to maintain the homeostasis of oxygen environment in vivo. The current research on hypoxic effect mainly focuses on the damage of hypoxia to cells and related adaptive mechanisms, and on the proliferation of hypoxia in cells. There are few studies on the effects of differentiation, especially in some physiological and pathological processes during embryogenesis and adulthood in vivo. Insufficient attention was paid to the occurrence of hypoxia in the local microenvironment. Stem cells, including bone marrow mesenchymal stem cells (MSCs), were cultured and differentiated in vitro. Under conditions. As a physiological stimulant in vivo, the effect of hypoxia on the proliferation and differentiation of MSCs cultured in vitro is still unclear. Human bone marrow mesenchymal stem cells (hMSCs) are currently the most promising adult stem cells in clinical application. The effects of hypoxia on proliferation and differentiation of hMSCs in vitro were studied. 1. Effects of hypoxia on proliferation of hMSCs in vitro To investigate the effect of hypoxia on the proliferation of hMSCs in vitro, the intermittent hypoxia of 3O 2 was observed by blood cell counter counting and flow cytometry, respectively. 10 / 10 mol/L / CoCl_2 / s / / /\\\. The effects of 200 渭 mol/L CoCl2) on the number of hMSCs and proliferation index were observed. The results showed that intermittent hypoxia had no significant effect on the number and proliferation index of hMSCs. The number and proliferation index of hMSCs were increased in the groups of persistent hypoxia, indicating that persistent hypoxia could promote the proliferation of hMSCs in vitro. It is suggested that persistent hypoxia, as a controllable factor in vitro, can regulate the proliferation of hMSCs in vitro, and has a certain significance for the clinical application of hMSCs. At the same time, gene microarray technique was used to analyze the changes of gene expression profile during hypoxia on hMSCs proliferation. The results showed that 282 genes were differentially expressed in 21329 genes. The effect of hypoxia on the proliferation of hMSCs was proved to be a complex process in which multiple genes were involved and five target genes were screened from the results of gene microarray. The expression changes of these five genes were detected by RT-PCR.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R329

【引證文獻】

相關(guān)碩士學(xué)位論文 前1條

1 馬洪巖;低氧對大鼠大腦皮層細胞凋亡及ROS、NO水平的影響[D];上海體育學(xué)院;2011年

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本文編號：1405708