| 鲍晶晶 葛欧洋 李璐 林艳 李晓曦 赵子建.Rheb 过表达在前体脂肪细胞株3T3-L1 分化中的作用[J].,2015,15(22):4217-4220 |
| Rheb 过表达在前体脂肪细胞株3T3-L1 分化中的作用 |
| The Function of Rheb Overexpression in 3T3-L1 Pre-adipocyteDifferentiation |
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| DOI： |
| 中文关键词: Rheb mTOR 3T3-L1 前脂肪细胞分化 |
| 英文关键词: Rheb mTOR 3T3-L1 pre-adipocyte |
| 基金项目:国家自然科学基金项目（81170780） |
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| 中文摘要: |
| 目的：利用前体脂肪细胞株3T3-L1 细胞观察mTOR（mammalian target of rapamycin）信号通路中上游调控因子Rheb（Ras
homolog enriched in brain）对其分化的影响。方法：利用高表达Rheb 的基因重组质粒转染前体脂肪细胞株，3T3-L1。通过蛋白质免
疫印迹实验鉴定质粒成功转染细胞后，诱导该细胞脂肪分化。予以分化第8 天的3T3-L1 细胞油红染色，并检测细胞内甘油三酯
的含量。另外，我们用Western blot 方法检测脂肪细胞特异性转录因子PPAR-r（Peroxisome proliferator-activated receptor-r）和
C/EBP-alpha（CCAAT-enhancer-binding protein-alpha）的表达情况来研究Rheb 在脂肪细胞分化过程中的作用。结果：我们成功构建了高表
达Rheb 的3T3-L1细胞株，发现高表达Rheb 后可以促进脂滴的生成，油红O 染色有显著区别，与对照组相比Rheb 高表达组的
三酰甘油含量明显升高(P<0.05)；C/EBP-alpha和PPAR-r等脂肪细胞特异性的转录因子蛋白表达量与对照组相比也均有升高（P<0.
05）。结论：Rheb 基因作为mTOR 通路上游调控因子，可以促进脂肪细胞的分化。 |
| 英文摘要: |
| Objective:To observe the effects of Rheb, an upstream regulating factor in mTOR (mammalian target of rapamycin)
signaling pathway, on the differentiation of pre-adipocytes, 3T3-L1.Methods:The plasmid pCAG-Insulator-Rheb was transfected into
mouse pre-adipocyte 3T3-L1 cells to overexpress Rheb. Western blot analysis was applied to verify the overexpression in 3T3-L1 cells.
On day 8 of differentiation, the 3T3-L1cells were stained with Oil Red O dye and analyzed for triglyceride contents in the 3T3-L1 cells.
Moreover, the expression of adipocyte differentiation marker genes, PPAR-r(Peroxisome proliferator-activated receptor-r) and C/EBP-alpha
(CCAAT-enhancer-binding protein-alpha), were examined by Western blot analysis. These parameters were used to assess the role of Rheb
in the differentiation of 3T3-L1 cells.Results:We successfully overexpressed Rheb in 3T3-L1 cells. Overexpressing Rheb promoted
adipogenesis and increased the content of triglyceride (P<0.05) judging by the elevated Oil Red O staining and expression of adipocyte
marker genes, PPAR-r and C/EBP-alpha(P<0.05).Conclusion:As the upstream regulatory factor of mTOR pathway, Rheb can promote
adipocyte differentiation. |
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