本文選題：骨髓間充質(zhì)干細胞 + 轉(zhuǎn)化生長因子β1　； 參考：《南京醫(yī)科大學(xué)》2008年碩士論文

【摘要】： 目的:探討TGF-β1對體外培養(yǎng)的骨髓間充質(zhì)干細胞(BMSCs)VEGF表達的影響 方法:(1)體外原代培養(yǎng)SD大鼠骨髓間充質(zhì)干細胞,每日觀察細胞生長狀態(tài),并鑒定。(2)將不同濃度的TGF-β1按不同的時間點作用于BMSCs,用ELISA方法觀察上清液中VEGF蛋白的表達,RT-PCR觀察VEGF mRNA的表達。(3)TGF-β1按1小時、2小時、3小時預(yù)處理BMSCs,觀察磷酸化Akt及磷酸化Erk的表達。(4)Akt通道阻斷劑LY294002、Erk通道阻斷劑U0126預(yù)處理1小時后,TGF-β1作用于BMSCs24小時,分別用ELISA和RT-PCR觀察VEGF蛋白及其mRNA表達情況。(5)LY294002和U0126預(yù)處理1小時后,TGF-β1作用于BMSCs,分別觀察1小時、2小時、3小時后,磷酸化Akt及磷酸化Erk1/2的表達。 結(jié)果:(1)成功培養(yǎng)出大鼠BMSCs,流式細胞儀鑒定細胞表面標記物CD45(-)CD90(+),本實驗中使用的細胞為第3-5代細胞。(2)TGF-β1能夠呈時間-濃度-依賴性促進BMSCs表達VEGF,并在蛋白和mRNA水平上驗證。(3)TGF-β1促進BMSCs表達VEGF的作用能夠被Akt通道特異性阻斷劑Ly 294002所拮抗,也能被ERK1/2的特異性阻斷劑U0126所阻斷,表明Akt和ERK1/2通道可能參與了這一過程。 結(jié)論:TGF-β1能夠在體外呈時間-濃度-依賴性促進大鼠BMSCs表達VEGF,并且這種促分泌作用可能是通過Akt和ERK1/2通道的激活來實現(xiàn)。
[Abstract]:Objective: to investigate the effect of TGF- 尾 1 on VEGF expression of bone marrow mesenchymal stem cells (BMSCs) cultured in vitro. (2) BMSCs were treated with different concentrations of TGF- 尾 _ 1 at different time points. The expression of VEGF protein in supernatant was observed by Elisa. (3) TGF- 尾 _ 1 was pretreated with BMSCs for 1 hour and 2 hours and 3 hours for phosphorylation of Akt and phosphoric acid. (4) Akt channel blocker LY294002Erk channel blocker U0126 pretreated for 1 hour, TGF- 尾 1 acted on BMSCs for 24 hours. (5) after pretreatment with LY294002 and U0126 for 1 hour, TGF- 尾 1 was applied to BMSCs. The expression of phosphorylated Akt and phosphorylated Erk 1 / 2 were observed after 1 hour and 2 hours and 3 hours, respectively. Results: (1) Rat BMSCs were successfully cultured, and the cell surface marker CD45 (-) CD90 (), was identified as the 3-5 passage cells by flow cytometry. (2) TGF- 尾 1 could promote the expression of VEGF in a time-concentration-dependent manner and at the protein and mRNA levels. (3) the effect of TGF- 尾 1 on the expression of Akt was antagonized by Ly 294002, a specific blocker of Akt channel. It can also be blocked by U0126, a specific inhibitor of ERK1 / 2, suggesting that Akt and ERK1 / 2 channels may be involved in this process. Conclusion: TGF- 尾 1 can promote the expression of VEGF in rat BMSCs in a time-concentration-dependent manner in vitro, and this effect may be mediated by the activation of Akt and ERK1 / 2 channels.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R329;R687.3

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