TGF-β1对大鼠BMSCs VEGF表达的影响及其机制

发布时间：2018-06-24 09:15

本文选题：骨髓间充质干细胞 + 转化生长因子β1　；参考：《南京医科大学》2008年硕士论文

【摘要】： 目的:探讨TGF-β1对体外培养的骨髓间充质干细胞(BMSCs)VEGF表达的影响方法:(1)体外原代培养SD大鼠骨髓间充质干细胞,每日观察细胞生长状态,并鉴定。(2)将不同浓度的TGF-β1按不同的时间点作用于BMSCs,用ELISA方法观察上清液中VEGF蛋白的表达,RT-PCR观察VEGF mRNA的表达。(3)TGF-β1按1小时、2小时、3小时预处理BMSCs,观察磷酸化Akt及磷酸化Erk的表达。(4)Akt通道阻断剂LY294002、Erk通道阻断剂U0126预处理1小时后,TGF-β1作用于BMSCs24小时,分别用ELISA和RT-PCR观察VEGF蛋白及其mRNA表达情况。(5)LY294002和U0126预处理1小时后,TGF-β1作用于BMSCs,分别观察1小时、2小时、3小时后,磷酸化Akt及磷酸化Erk1/2的表达。结果:(1)成功培养出大鼠BMSCs,流式细胞仪鉴定细胞表面标记物CD45(-)CD90(+),本实验中使用的细胞为第3-5代细胞。(2)TGF-β1能够呈时间-浓度-依赖性促进BMSCs表达VEGF,并在蛋白和mRNA水平上验证。(3)TGF-β1促进BMSCs表达VEGF的作用能够被Akt通道特异性阻断剂Ly 294002所拮抗,也能被ERK1/2的特异性阻断剂U0126所阻断,表明Akt和ERK1/2通道可能参与了这一过程。结论:TGF-β1能够在体外呈时间-浓度-依赖性促进大鼠BMSCs表达VEGF,并且这种促分泌作用可能是通过Akt和ERK1/2通道的激活来实现。
[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.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R329;R687.3

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