【摘要】：前言 治療性血管新生(therapeutic angiogenesis)是通過血管生成促進(jìn)因子的直接應(yīng)用,特異性轉(zhuǎn)基因治療及細(xì)胞治療以促進(jìn)缺血組織血管生成,改善心肌血供為目的的治療。缺血性心臟病的治療目的是擴(kuò)張冠狀動(dòng)脈,促進(jìn)血管新生,改善心肌血供。多種血管生成促進(jìn)因子參與心臟新生血管形成,用于治療性血管新生。其中,血管內(nèi)皮生長(zhǎng)因子(vascular endothelial growth factor,VEGF)和堿性成纖維細(xì)胞生長(zhǎng)因子(basic fibroblast growth factor,bFGF)是最重要的并且研究較多的兩種血管生成促進(jìn)因子。本實(shí)驗(yàn)在體外培養(yǎng)大鼠心臟微血管內(nèi)皮細(xì)胞(cardiac microvascular endothelial cell,CMEC),并應(yīng)用Matrigel建立誘導(dǎo)血管腔樣結(jié)構(gòu)形成的體外培養(yǎng)體系,研究血管生成促進(jìn)因子bFGF對(duì)誘導(dǎo)心臟微血管內(nèi)皮細(xì)胞形成管腔樣結(jié)構(gòu)的影響,和bFGF對(duì)內(nèi)皮細(xì)胞VEGF受體Flt-1的作用,進(jìn)而探討bFGF促進(jìn)血管形成過程的作用機(jī)制。 方法 首先,參考Nishida等的實(shí)驗(yàn)方法,分離、培養(yǎng)大鼠心臟微血管內(nèi)皮細(xì)胞。根據(jù)不同的實(shí)驗(yàn)因素濃度設(shè)計(jì)成濃度效應(yīng)組:不同濃度bFGF(5,10,20,40ng/ml)和正常對(duì)照組(無血清DMEM培養(yǎng)液),分別作用24小時(shí)。應(yīng)用Matrigel誘導(dǎo)管腔樣結(jié)構(gòu)形成的體外培養(yǎng)體系,檢測(cè)不同濃度bFGF作用后,心臟微血管內(nèi)皮細(xì)胞形成管腔的情況。RT-PCR檢測(cè)各組血管內(nèi)皮細(xì)胞的Flt-1mRNA表達(dá)量。最后統(tǒng)計(jì)分析結(jié)果。 結(jié)果 實(shí)驗(yàn)結(jié)果表明,應(yīng)用Matrigel可誘導(dǎo)心臟微血管內(nèi)皮細(xì)胞管腔樣結(jié)構(gòu)
[Abstract]:The purpose of therapeutic angiogenesis (therapeutic angiogenesis) is to promote angiogenesis in ischemic tissues through the direct application of angiogenic promoters, specific transgene therapy and cell therapy. To improve myocardial blood supply for the purpose of treatment. The aim of treatment of ischemic heart disease is to dilate coronary artery, promote angiogenesis and improve myocardial blood supply. A variety of angiogenesis promoters are involved in cardiac neovascularization and are used for therapeutic angiogenesis. Among them, vascular endothelial growth factor (vascular endothelial growth factor,VEGF) and basic fibroblast growth factor (basic fibroblast growth factor,bFGF) are the two most important and studied angiogenesis promoting factors. In this study, rat cardiac microvascular endothelial cells (cardiac microvascular endothelial cell,CMEC) were cultured in vitro, and Matrigel was used to establish an in vitro culture system to induce the formation of vascular lumen-like structure. To study the effect of angiogenesis promoting factor bFGF on the formation of lumen-like structure in cardiac microvascular endothelial cells and the effect of bFGF on VEGF receptor Flt-1 in endothelial cells, and to explore the mechanism of bFGF promoting angiogenesis. Methods firstly, the microvascular endothelial cells of rat heart were isolated and cultured according to the experimental methods of Nishida et al. Different concentration of bFGF (5,10,20,40 ng / ml) and normal control group (serum-free DMEM medium) were designed for 24 hours respectively according to the concentration of the experimental factors and the concentration of the control group (5 ng / ml, 10 ng / ml, 20 ng / ml, 40 ng / ml). In vitro culture system induced by Matrigel to induce lumen-like structure was used to detect the formation of lumen in cardiac microvascular endothelial cells treated with different concentrations of bFGF, and Flt-1mRNA expression in vascular endothelial cells in each group was measured by RT-PCR. Finally, statistical analysis of the results. Results the results showed that Matrigel could induce the lumen-like structure of cardiac microvascular endothelial cells.
【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R363.2

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