文章摘要
张 许,林海燕,尹 业,韩 晓,程 光.3T3-L1前脂肪细胞分化过程中自噬的发生与线粒体数目的改变[J].南京医科大学学报,2015,(12):1675~1679
3T3-L1前脂肪细胞分化过程中自噬的发生与线粒体数目的改变
Appearence of autophagy and changes of mitochondria number in the process of 3T3-L1 preadipocyte differentiation
投稿时间:2015-09-19  
DOI:10.7655/NYDXBNS20151201
中文关键词: 自噬  脂肪细胞分化  线粒体
英文关键词: autophagy  adipocyte differentiation  mitochondria
基金项目:国家自然科学基金资助(H0704)
作者单位
张 许 南京医科大学江苏省人类功能基因组学重点实验室,江苏 南京 210029 
林海燕 南京医科大学江苏省人类功能基因组学重点实验室,江苏 南京 210029 
尹 业 南京医科大学江苏省人类功能基因组学重点实验室,江苏 南京 210029 
韩 晓 南京医科大学江苏省人类功能基因组学重点实验室,江苏 南京 210029 
程 光 南京绿叶制药有限公司长效和靶向制剂国家重点实验室,江苏 南京 210061 
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中文摘要:
      目的:探索3T3-L1前脂肪细胞分化过程中自噬的发生与线粒体数目改变的关系。方法:培养3T3-L1前脂肪细胞,利用电子显微镜技术观察细胞分化过程中自噬小泡的形成与变化?线粒体数目的改变,同时运用Western blot检测NRF1?mtTFA的表达。结果:电镜结果显示:在3T3-L1 前脂肪细胞分化过程中,自噬小泡出现在细胞分化的第2天,随后迅速增加,第4天自噬小泡的面积达到最大,第6天开始消失;线粒体数目在分化第2天显著下降,第4天开始逐渐增加,到第6天超过分化前的水平并在第8天恢复;在分化过程中的自噬小泡内,有被吞噬的线粒体。Western blot结果则显示调控线粒体合成的转录因子NRF1?mtTFA的表达在分化第2天显著下降,第4天迅速升高,至分化末期再次下降。结论:3T3-L1前脂肪细胞分化过程的早中期形成了大量的自噬小泡,包括一些线粒体自噬,这些线粒体自噬的发生可能是分化早期线粒体数目减少的原因之一。
英文摘要:
      Objective:To explore the relationship between the changes of autophagy and the number of mitochondria in the process of the differentiation of 3T3-L1 cells. Methods:(1)3T3-L1 preadipocytes were cultured and then induced by media mixed with insulin,DEX,IBMX. Lipid droplets were observed on the fifth day. The peroxisome proliferation activated receptor γ(PPARγ)and CAAT Enhancer Binding Proteins α(CEBPα)protein were detected by Western blotting. (2)Autophagosome pictures were taken by transmission electron microscope (TEM)and LC3II level was detected by Western blotting. Results:The TEM showed that during the differentiation of 3T3-L1 preadipocyte,autophagy vesicles appeared on the second day,extended to greater range of cytoplasm on the fourth day,then gradually disappeared from the sixth day. While the expression of LC3Ⅱ began to increase in the second day, reached the highest on the fourth day,and then gradually declined. Furthermore,with the change of autophagy vesicles during the differentiation,the number of mitochondria increased after a decrease. Conclusion:There are a lot of autophagy vesicles in the early and middle stage of the differentiation of 3T3-L1,including some mitophagy,which may be one of the reasons for the decrease of the number of mitochondria in early stage of differentiation.
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