本文關(guān)鍵詞： 間質(zhì)干細(xì)胞 基因 GATA 血紅素加氧酶- 心肌分化　出處：《解放軍醫(yī)學(xué)雜志》2017年04期 　論文類型：期刊論文

【摘要】：目的探討經(jīng)人血紅素加氧酶1(hHO-1)基因和GATA-4基因聯(lián)合修飾的大鼠骨髓間充質(zhì)干細(xì)胞(BMSCs)在缺血缺氧環(huán)境中抗凋亡及體外分化為心肌細(xì)胞樣細(xì)胞的能力是否優(yōu)于hHO-1或GATA-4單基因修飾的BMSCs。方法采用全骨髓貼壁法分離、培養(yǎng)BMSCs并進(jìn)行鑒定,重組腺病毒轉(zhuǎn)染BMSCs,Western blotting法確定基因表達(dá)的最佳時(shí)間;以缺氧無血清條件模擬體內(nèi)缺血缺氧微環(huán)境培養(yǎng)基因修飾的BMSCs,采用CCK-8試劑盒和臺盼藍(lán)染色法分別檢測缺血缺氧培養(yǎng)12、24、48、72h細(xì)胞存活率,流式細(xì)胞術(shù)檢測缺血缺氧培養(yǎng)24h細(xì)胞凋亡情況,Western blotting及細(xì)胞免疫熒光法檢測特異性心肌肌鈣蛋白I(cTnI)表達(dá)情況。結(jié)果缺血缺氧培養(yǎng)12、24、48、72h的hHO-1+GATA-4組細(xì)胞生存率均高于hHO-1組(P0.05)和GATA-4組(P0.05)。缺血缺氧培養(yǎng)24h的hHO-1+GATA-4組細(xì)胞凋亡率低于hHO-1組(P0.05)和GATA-4組(P0.05);GATA-4組與hHO-1+GATA-組的c Tn I表達(dá)差異無統(tǒng)計(jì)學(xué)意義。結(jié)論 hHO-1與GATA-4聯(lián)合修飾可明顯增強(qiáng)缺血缺氧BMSCs的存活;與GATA-4單基因修飾相比,聯(lián)合修飾并沒有增強(qiáng)BMSCs向心肌細(xì)胞分化的能力。
[Abstract]:Objective to investigate the ability of rat bone marrow mesenchymal stem cells modified by human heme oxygenase 1hHO-1 gene and GATA-4 gene to inhibit apoptosis and differentiate into cardiomyocyte-like cells in vitro under ischemia and hypoxia environment. Methods the ability of BMSCs modified by human heme oxygenase (HO-1) gene and GATA-4 gene to differentiate into cardiomyocyte-like cells in vitro was better than that of hHO-1 or GATA-4 alone. Methods whole bone marrow adherent method was used to isolate BMSCs. BMSCs was cultured and identified. The optimal time for gene expression was determined by Western blotting method. CCK-8 kit and Trypan blue staining were used to detect the survival rate of cells cultured with anoxia and anoxia for 72 h. Flow cytometry (FCM) was used to detect the apoptosis of hHO-1 GATA-4 cells cultured with ischemia and hypoxia for 24 h and the expression of specific cardiac troponin I cTnI by immunofluorescence assay. Results the cell survival rate of hHO-1 GATA-4 group was higher than that of hHO-1 group at 72 h after ischemia and hypoxia culture. The apoptosis rate of hHO-1 GATA-4 group was lower than that of hHO-1 group (P 0.05) and that of GATA-4 group was not significantly different from that of hHO-1 GATA-4 group and hHO-1 GATA-group. Conclusion hHO-1 combined with GATA-4 modification can significantly enhance the survival of hypoxic ischemic BMSCs. Compared with GATA-4 single gene modification, combined modification did not enhance the ability of BMSCs to differentiate into cardiomyocytes.
【作者單位】： 南方醫(yī)科大學(xué)研究生院;郴州市第一人民醫(yī)院高通量分子診斷技術(shù)國家&地方聯(lián)合工程實(shí)驗(yàn)室;解放軍305醫(yī)院中心實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金(30971235) 軍隊(duì)“十二五”醫(yī)學(xué)科研基金(2013XL010)~~
【分類號】：R542.22
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本文編號：1535166