上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

蛋白激酶MPK-1通过DAF-16调控FAT-7抑制脂质累积

韩三峰，黄黎莹，王立顺   

  1. 上海交通大学 医学院附属瑞金医院分子医学中心，上海 200025
  • 出版日期:2013-07-28 发布日期:2013-08-22
  • 通讯作者: 王立顺, 电子信箱: jywangls@shsmu.edu.cn。
  • 作者简介:韩三峰（1986—），男，硕士; 电子信箱: lovey1123@yahoo.cn。

Protein kinase MPK-1 inhibits fat accumulation via regulating FAT-7 by DAF-16

HAN San-feng, HUANG Li-ying, WANG Li-shun   

  1. Center for Molecular Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2013-07-28 Published:2013-08-22

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摘要：

目的 探讨蛋白激酶MPK-1对脂质代谢的影响及可能的作用机制。方法 用秀丽线虫（C.elegans）作为模式生物，采用RNA干扰（RNAi）技术定向干扰mpk-1，sbp-1、daf-2基因表达，油红O和苏丹黑染色观察线虫脂质含量的变化，洗脱脂肪进行定量分析确定脂肪含量的增减。采用RNAi技术结合对线虫体内特定位置绿色荧光蛋白表达变化的观察，分析MPK-1影响脂肪累积的可能机制。结果 干扰mpk-1基因表达后，线虫体内脂肪含量明显增加。荧光显微镜观察发现：脂质合成酶fat-7∷gfp线虫干扰mpk-1基因表达后，荧光增强；表达调控fat-7的转录因子DAF-16荧光蛋白的线虫品系中干扰mpk-1基因表达，可促进DAF-16向核内转移。结论 蛋白激酶MPK-1可能通过DAF-16调控FAT-7的活性，抑制脂肪的合成代谢，减少脂质累积。

关键词: 蛋白激酶, MPK-1, 脂质, RNA干扰, 秀丽隐杆线虫

Abstract:

Objective To investigate the effect of protein kinase MPK-1 on lipid metabolism, and explore the underlying mechanism. Methods C.elegans was served as model organism, and the expression of mpk-1, sbp-1 and daf-2 genes was interfered with RNA interference (RNAi). The fat contents in C.elegans were observed by Oil-red O staining and Sudan black staining, and were quantified after washing off the dye. Then, the mechanism of effect of MPK-1 on fat accumulation was determined by the combination of RNAi knockdown technique and the green fluorescence change in a specified location. Results The fat content significantly increased after interference of mpk-1 gene expression. Fluorescence microscopy revealed that fluorescence enhancement was detected after interference of mpk-1 gene expression in C.elegans with expression of lipogenic enzymes fat-7∷gfp, and intranuclear transference of DAF-16 was promoted after interference of mpk-1 gene expression in C.elegans with expression of DAF-16 fluorescin regulating fat-7. Conclusion Protein kinase MPK-1 may inhibit FAT-7 activity through DAF-16, and reduce the fat accumulation.

Key words: protein kinase, MPK-1, fat, RNA interference, C.elegans