本文選題：干細(xì)胞 + 間質(zhì)干細(xì)胞　； 參考：《中國(guó)組織工程研究》2017年25期

【摘要】：背景:目前發(fā)現(xiàn)低氧誘導(dǎo)因子1α(hypoxia inducible factor 1,HIF1α)可提高組織和細(xì)胞在缺血環(huán)境下繼續(xù)生存的能力,在促進(jìn)血管再生方面有重要意義。目的:構(gòu)建三點(diǎn)突變型HIF1α聯(lián)合示蹤因子hrGFP雙基因重組腺病毒載體Ad-CMV-HIF1α~(mu)-IRES-hrGFP-1,研究其對(duì)骨髓間充質(zhì)干細(xì)胞增殖的影響。方法:將第3代骨髓間充質(zhì)干細(xì)胞分組培養(yǎng),實(shí)驗(yàn)組以突變基因Ad-CMV-HIF1α~(mu)-IRES-hrGFP-1病毒液轉(zhuǎn)染細(xì)胞,陽(yáng)性對(duì)照組以未突變基因Ad-CMV-HIF1α-IRES-hrGFP-1病毒液轉(zhuǎn)染細(xì)胞,陰性對(duì)照組以Ad-CMV-IRES-hrGFP-1空病毒液轉(zhuǎn)染細(xì)胞,空白對(duì)照組不轉(zhuǎn)染任何病毒液。轉(zhuǎn)染72 h后,進(jìn)行HIF1α基因mRNA及蛋白檢測(cè);轉(zhuǎn)染24,48,72,96,120 h,MTT法檢測(cè)細(xì)胞增殖。結(jié)果與結(jié)論:(1)HIF1α基因mRNA檢測(cè):實(shí)驗(yàn)組、陽(yáng)性對(duì)照組HIF1α基因mRNA表達(dá)高于陰性對(duì)照組、空白對(duì)照組(P0.05),其余組間兩兩比較差異無(wú)顯著性意義;(2)HIF1α蛋白檢測(cè):實(shí)驗(yàn)組細(xì)胞中HIF1α蛋白表達(dá)高于陽(yáng)性對(duì)照組、陰性對(duì)照組、空白對(duì)照組(P0.05),后3組間比較差異無(wú)顯著性意義;(3)細(xì)胞增殖:陽(yáng)性對(duì)照組與空白對(duì)照組細(xì)胞生長(zhǎng)速度均較慢,兩組間相比較無(wú)顯著性意義;實(shí)驗(yàn)組細(xì)胞增殖程度高于陽(yáng)性對(duì)照組、空白對(duì)照組(P0.05);(4)結(jié)果表明,重組腺病毒介導(dǎo)的三點(diǎn)突變型HIF1α聯(lián)合hrGFP基因轉(zhuǎn)染骨髓間充質(zhì)干細(xì)胞后,不僅可在常氧條件下持續(xù)表達(dá)目的蛋白HIF1α,同時(shí)也可促進(jìn)骨髓間充質(zhì)干細(xì)胞的增殖。
[Abstract]:Background: at present, it is found that hypoxia inducible factor 1 alpha (hypoxia inducible factor 1, HIF1 a) can improve the ability of tissue and cells to continue to survive in the ischemic environment, and it is of great significance in promoting vascular regeneration. Objective: to construct the three point mutant HIF1 alpha combined with hrGFP double gene recombinant adenovirus vector Ad-CMV-HIF1 a ~ (MU) -IRES-hrGFP-. 1, study its effect on the proliferation of bone marrow mesenchymal stem cells. Methods: the third generation bone marrow mesenchymal stem cells were grouped in groups. The experimental group transfected cells with the mutant gene Ad-CMV-HIF1 alpha ~ (MU) -IRES-hrGFP-1 virus solution. The positive control group transfected the cells with the Ad-CMV-HIF1 alpha -IRES-hrGFP-1 virus of the non mutant gene, and the negative control group was Ad-CMV-IRES-hrG FP-1 empty virus fluid transfected cells, blank control group did not transfect any virus fluid. After transfection of 72 h, HIF1 alpha gene mRNA and protein detection, 24,48,72,96120 h, MTT method to detect cell proliferation. Results and conclusion: (1) HIF1 a gene mRNA detection: experimental group, mRNA control group HIF1 alpha gene mRNA higher than the negative control group, the blank control group (P0) .05), there was no significant difference between the 22 other groups. (2) HIF1 alpha protein detection: the expression of HIF1 alpha protein in the experimental group was higher than that in the positive control group, the negative control group and the blank control group (P0.05), and there was no significant difference between the 3 groups. (3) the growth rate of the cells in the positive control group and the blank control group were both slower and two groups. The proliferation of cells in the experimental group was higher than that in the positive control group and the blank control group (P0.05). (4) the results showed that the recombinant adenovirus mediated three point mutant HIF1 alpha combined with hrGFP gene transfected to bone marrow mesenchymal stem cells could not only express the target protein HIF1 alpha continuously under the normal oxygen strip, but also promote the bone marrow. Proliferation of mesenchymal stem cells.
【作者單位】： 錦州醫(yī)科大學(xué)附屬第一醫(yī)院整形外科;錦州醫(yī)科大學(xué)附屬第一醫(yī)院臨床生物樣本中心;
【基金】：遼寧醫(yī)學(xué)院校長(zhǎng)基金——臨床醫(yī)學(xué)建設(shè)專(zhuān)項(xiàng)基金項(xiàng)目資助(XZJJ20140215)~~
【分類(lèi)號(hào)】：R329.2

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