| 李金存 郑秉暄 王超 高星 叶菁.雷帕霉素对小鼠原代肝细胞脂滴形态和脂滴表面蛋白表达的影响[J].,2014,14(27):5220-5224 |
| 雷帕霉素对小鼠原代肝细胞脂滴形态和脂滴表面蛋白表达的影响 |
| Effects of Rapamycin on the Morphology of Lipid Droplets and theExpression of Lipid Droplet-associated Proteins in Mouse PrimaryHepatocytes |
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| DOI： |
| 中文关键词: 雷帕霉素 脂滴 脂滴表面蛋白 mTOR |
| 英文关键词: Rapamycin Lipid droplets Lipid droplet-associated proteins mTOR |
| 基金项目:国家自然科学基金项目（81101752；81101711） |
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| 中文摘要: |
| 目的：探讨雷帕霉素（Rapamycin）对小鼠原代肝细胞脂滴形态和脂滴表面蛋白表达的影响。方法：采用胶原酶灌注方法分离和培养小鼠原代肝细胞，采用100 uM油酸诱导肝细胞内脂肪的合成。采用0、10、20、50 滋M 的雷帕霉素处理肝细胞12 hr 后，利用中性脂肪染料Bodipy 493/503 对肝细胞内的脂滴进行染色，荧光显微镜下观察细胞脂滴形态和数量。定量试剂盒检测细胞内甘
油三酯（TG）的含量利用Western blot检测不同浓度雷帕霉素处理的小鼠原代肝细胞脂滴表面蛋白ADRP的表达水平。结果：成功分离和培养了小鼠原代肝细胞，使用油酸处理能够明显增加原代肝细胞内脂滴的数量。随着体外雷帕霉素处理浓度的增加，荧光显微镜下观察发现原代肝细胞内脂滴的数量呈现明显的下降趋势，甘油三酯的含量也呈见明确的下降趋势，在20 uM 浓度下就表现出显著性差异。Western blot 结果显示雷帕霉素能够在抑制肝细胞内脂肪储积的同时降低脂滴表面蛋白ADRP的表达水平，并且随着雷帕霉素处理浓度的增加，其对ADRP表达的抑制越明显。结论：雷帕霉素能够抑制肝细胞内中性脂肪的储积，同时
降低脂滴表面蛋白ADRP的表达水平。也间接说明了mTOR 信号通路能够影响肝细胞内脂肪的储积，也为脂肪肝的防治提供了
一个新的实验基础 |
| 英文摘要: |
| Objective:To investigate the effects of different concentration of rapamycin on the morphology of lipid droplets(LD)
and the expression of lipid droplet-associated proteins in mouse primary hepatocytes.Methods:The primary hepatocytes of mice were
isolated and cultured using collagenase perfusion. 200 uMoleic acid (0A) was used to stimulate and induce the production of the lipid in
the hepatocytes and then 0 (control), 10, 20, 50 uM rapamycin were added respectively. 12 hours later, Bodipy 493/503 staining was
used to observe the morphology and amount of intracellular LD under fluorescence microscope. The Folch method and triglyceride
quantitative kit were used to extract the total intracellular lipids and determine the TG contents. Furthermore, the expression of lipid
droplet associated proteins (ADRP) was analyzed by Western Blot.Results:The mouse primary hepatocytes were isolated and cultured
successfully. Oleic acid can increase the amount of intracellular LD. Along with the increase of treated concentration of rapamycin, the
number of LDs and the content of TG in hepatocytes were gradually decreased. In addition, rapamycin could reduce the expression levels
of ADRP.Conclusion:Rapamycin can inhibit intracellular lipid accumulation and decrease the expression levels of ADRP, suggesting
the involvement of mTOR signaling pathway in intracellular lipid accumulation and providing the mechanism for the important role of
mTOR in the cellular energy metabolism. |
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