本文關(guān)鍵詞：腎上腺髓質(zhì)素基因轉(zhuǎn)染的骨髓間充質(zhì)干細胞缺氧共培養(yǎng)對新生大鼠心肌細胞凋亡的影響　出處：《福建醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 骨髓間充質(zhì)干細胞 乳鼠心肌細胞 腎上腺髓質(zhì)素 凋亡 共培養(yǎng)

【摘要】：[目的]研究腎上腺髓質(zhì)素（ADM）基因轉(zhuǎn)染的骨髓間充質(zhì)干細胞（MSCs）缺氧共培養(yǎng)對新生大鼠心肌細胞凋亡的影響。 [方法]貼壁法體外分離、擴增培養(yǎng)大鼠骨髓間充質(zhì)干細胞。用x-gal染色測含腎上腺髓質(zhì)素基因的重組腺病毒(Ad-ADM)對骨髓間充質(zhì)干細胞感染率。乳鼠心肌細胞分離、純化、培養(yǎng)與鑒定。乳鼠心肌細胞缺氧、無血清條件下培養(yǎng)3、6、12、24小時并檢測其凋亡率。用Transwell小室缺氧、無血清條件下共培養(yǎng)骨髓間充質(zhì)干細胞和乳鼠心肌細胞，24小時后檢測共培養(yǎng)體系上清液中腎上腺髓質(zhì)素含量并檢測乳鼠心肌細胞凋亡率。實驗分組為：Ad-ADM轉(zhuǎn)染共培養(yǎng)組，Ad-Null轉(zhuǎn)染共培養(yǎng)組，wildtype共培養(yǎng)組，同時設(shè)置單獨乳鼠心肌細胞缺氧、無血清培養(yǎng)組。 [結(jié)果]重組腺病毒對骨髓間充質(zhì)干細胞的轉(zhuǎn)染率與病毒感染復(fù)數(shù)（MOI）具有量效關(guān)系，MOI為150時細胞的感染率達95.4%。在缺氧、無血清培養(yǎng)條件下，乳鼠心肌細胞在缺氧3h、6h、12h、24h的凋亡率逐步升高，各時間點凋亡率差異具有統(tǒng)計學(xué)意義（P＜0.001）。三組共培養(yǎng)體系中乳鼠心肌細胞凋亡率明顯低于單獨乳鼠心肌細胞缺氧培養(yǎng)組（P＜0.05）；Ad-ADM轉(zhuǎn)染共培養(yǎng)組乳鼠心肌細胞凋亡率低于空載體轉(zhuǎn)染、共培養(yǎng)組和未轉(zhuǎn)染、共培養(yǎng)組（P＜0.05）。空載體轉(zhuǎn)染、共培養(yǎng)組和未轉(zhuǎn)染、共培養(yǎng)組相比，，乳鼠心肌細胞凋亡差異未達到統(tǒng)計學(xué)意義。腎上腺髓質(zhì)素基因轉(zhuǎn)染共培養(yǎng)組的上清液中腎上腺髓質(zhì)素分泌量高于空載體轉(zhuǎn)染共培養(yǎng)組和未轉(zhuǎn)染共培養(yǎng)組（P＜0.05）�？蛰d體轉(zhuǎn)染共培養(yǎng)組與未轉(zhuǎn)染共培養(yǎng)組相比，腎上腺髓質(zhì)素差異未達到統(tǒng)計學(xué)意義。 [結(jié)論]與骨髓間充質(zhì)干細胞共培養(yǎng)可以降低缺氧、無血清培養(yǎng)條件下乳鼠心肌細胞的凋亡率。與未轉(zhuǎn)染的骨髓間充質(zhì)干細胞相比，在缺氧、無血清培養(yǎng)條件下（24小時），腎上腺髓質(zhì)素基因轉(zhuǎn)染可以增強共培養(yǎng)對乳鼠心肌細胞的凋亡的影響，這可能與腎上腺髓質(zhì)素表達增加有關(guān)。
[Abstract]:[Objective] to study the effect of hypoxia co-culture of bone marrow mesenchymal stem cells (MSCs) transfected with adrenal medullary hormone (ADM) gene on cardiomyocyte apoptosis in neonatal rats. [Methods] the adherent method was isolated in vitro. Bone marrow mesenchymal stem cells (BMSCs) were amplified and cultured in rats. The infection rate of the recombinant adenovirus Ad-ADM containing adrenomedullin gene was detected by x-gal staining. Purification, culture and identification. Neonatal rat cardiomyocytes were exposed to hypoxia and cultured for 24 hours without serum. The apoptosis rate was measured by Transwell chamber hypoxia. Bone marrow mesenchymal stem cells and neonatal cardiomyocytes were co-cultured in serum-free medium. After 24 hours, the content of adrenomedullin in supernatant of co-culture system and the rate of cardiomyocyte apoptosis in neonatal rats were detected. Wildtype co-culture group, at the same time set up isolated neonatal rat cardiomyocytes hypoxia, serum-free culture group. [Results: the transfection efficiency of recombinant adenovirus on bone marrow mesenchymal stem cells was dose-dependent and the infection rate was 95.4 when moi was 150. Under the condition of serum-free culture, the apoptosis rate of neonatal rat cardiomyocytes increased gradually after hypoxia for 3 h, 6 h and 12 h for 24 h. There was significant difference in apoptosis rate at different time points (P < 0.001). The apoptosis rate of neonatal rat cardiomyocytes in the co-culture system was significantly lower than that in hypoxic culture group (P < 0.05). The apoptosis rate of neonatal rat cardiomyocytes in Ad-ADM transfection group was lower than that in empty vector transfection group, and the co-culture group and non-transfection group (P < 0.05), empty vector transfection, co-culture group and non-transfection group. Compared with co-culture group. The amount of adrenomedullin secreted in the supernatant of the co-culture group was higher than that in the empty vector co-culture group and the untransfected co-culture group. P < 0.05. The empty vector transfection co-culture group was compared with the non-transfection co-culture group. The difference of adrenomedullin was not statistically significant. [Conclusion: Co-culture with bone marrow mesenchymal stem cells can reduce the apoptosis rate of neonatal rat cardiomyocytes in anoxic and serum-free culture, compared with untransfected bone marrow mesenchymal stem cells. In serum-free culture for 24 hours, adrenomedullin gene transfection could enhance the effect of co-culture on cardiomyocyte apoptosis in neonatal rats, which may be related to the increase of adrenomedullin expression.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329

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