【摘要】：背景:腎上腺髓質(zhì)素基因轉(zhuǎn)染能增強骨髓間充質(zhì)干細胞在缺血、缺氧環(huán)境下的抗凋亡能力,但其機制尚未完全明確。目的:觀察腎上腺髓質(zhì)素對骨髓間充質(zhì)干細胞缺血、缺氧時Bax、Bcl-2和Caspase-3等凋亡相關調(diào)節(jié)蛋白表達的影響。方法:貼壁法分離、培養(yǎng)、純化SD大鼠骨髓間充質(zhì)干細胞,缺氧、無血清培養(yǎng)0,3,6,9,12 h后用Western blot測定Bax、Bcl-2、Caspase-3蛋白的表達以確定最佳缺氧時間。采用缺氧6 h為時間點,根據(jù)缺氧、無血清培養(yǎng)前是否進行腎上腺髓質(zhì)素預處理分為對照組(未加入腎上腺髓質(zhì)素)及腎上腺髓質(zhì)素不同質(zhì)量濃度組(1,10,100μg/L),然后采用Western blot測定Bax、Bcl-2、Caspase-3蛋白的表達。結(jié)果與結(jié)論:1無血清培養(yǎng)條件下缺氧0,3,6,9,12 h后骨髓間充質(zhì)干細胞Bax、Bcl-2表達均增加(P0.05),缺氧6 h時Bax/Bcl-2比值、caspase-3蛋白達到最小(P0.05)。2缺氧6 h時,與1,10μg/L腎上腺髓質(zhì)素組相比,100μg/L腎上腺髓質(zhì)素組表達Bax蛋白、Caspase-3蛋白最低,Bax/Bcl-2比值最低,而表達Bcl-2蛋白最高(P0.05)。結(jié)果表明腎上腺髓質(zhì)素能降低無血清、缺氧條件下骨髓間充質(zhì)干細胞Bax/Bcl-2比值、Caspase-3蛋白表達,該效應存在劑量依賴性。
[Abstract]:Background: adrenomedullin gene transfection can enhance the anti-apoptosis ability of bone marrow mesenchymal stem cells in ischemic and hypoxic environment, but its mechanism is not completely clear. Aim: to investigate the effects of adrenomedullin on the expression of apoptosis-related regulatory proteins such as Bax,Bcl-2 and Caspase-3 during ischemia and hypoxia of bone marrow mesenchymal stem cells. Methods: bone marrow mesenchymal stem cells (BMSCs) of SD rats were isolated, cultured and purified by adherent method. After hypoxia and serum-free culture for 12 h, the expression of Bax,Bcl-2,Caspase-3 protein was determined by Western blot to determine the best hypoxia time. According to whether adrenomedullin pretreatment was performed before anoxia and serum-free culture for 6 h, the rats were divided into two groups: control group (without adrenomedullin) and adrenomedullin with different concentration (1 10 100 渭 g / L). Then Western blot was used to detect the expression of Bax,Bcl-2,Caspase-3 protein. Results and conclusion: 1 the expression of Bax,Bcl-2 in bone marrow mesenchymal stem cells increased after 12 h hypoxia (P0.05), and the Bax/Bcl-2 ratio increased at 6 h after hypoxia. Caspase-3 protein reached the minimum (P0.05). 2 at 6 h hypoxia, compared with 1 渭 g / L adrenomedullin group, 100 渭 g / L adrenomedullin group expressed Bax protein, Caspase-3 protein was the lowest, and Bax/Bcl-2 ratio was the lowest. The expression of Bcl-2 protein was the highest (P0.05). The results showed that adrenomedullin could decrease the ratio of Bax/Bcl-2 and the expression of Caspase-3 protein in cultured bone marrow mesenchymal stem cells in a dose-dependent manner.
【作者單位】： 福建省立醫(yī)院北院福建省老年醫(yī)院心內(nèi)科;福建醫(yī)科大學附屬第一醫(yī)院心內(nèi)科高血壓研究所;
【基金】：福建省衛(wèi)生廳青年基金(2011-1-15,2011-1-32) 福建省自然科學基金(2014J01433)~~
【分類號】：R329.2

【參考文獻】

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