目的 鉴定CIDEC的表达能够被PPARγ和PGC1α共激活,检测其在脂肪代谢中的重要调控作用.方法 构建5删切和顺式原件突变的启动子,运用HepG2细胞中双荧光报告基因实验检测PPARγ和PGC1α对CIDEC的共激活作用并确定顺式作用元件.运用PPARγ特异激动剂piogiltazone刺激3T3-L1细胞检测CIDEC的mRNA水平.在3T3-L1细胞系中过表达CIDEC,检测脂肪合成及能量代谢相关基因的mRNA水平.通过腺病毒感染在小鼠肝原代细胞中过表达CIDEC,检测肝脏细胞中脂肪变化.结果 PPARγ和PGC1α能够显著激活CIDEC的启动子.过表达CIDEC后,脂肪细胞中脂类合成相关基因FAS上调(3.47±0.17)倍(P<0.05),ACC上调(3.95±0.57)倍(P<0.05),在肝原代细胞中CIDEC促进脂滴的生成.结论 CIDEC过表达能够被PPARγ促进并且被PGC1α共激活,在体内起到促进脂肪积累的作用.%Objective To identify the regulation of PPARγand PGC1α on CIDEC transcription and the function of CIDEC in adipose metabolism. Methods Reconstructe CIDEC promoter reporters including a series of 5'-deletions and cis-element mutations. Dual-luciferase assay was performed to screen PPARγ binding sites in HepG2 cells.Pioglitazone, the PPARγagonist, was used in 3T3-L1 cells to detect the CIDEC mRNA level. Adenoviral CIDEC was constructed and over-expressed CIDEC gene through adenovirus in primary hepatocytes to examine the effect of CIDEC on liver adipose metabolism. Results PPARγ and PGC1α can stimulate CIDEC transcription obviously.Over-expressing CIDEC stimulate gene FAS 3. 47 ±0. 17 (P ＜0. 05) and ACC 3. 95 ±0. 57 (P ＜0. 05), and adipose synthesis in hepatocytes. Conclusion Transcription of CIDEC can be activated by PPARγ and its coactivitor PGClα. This activation results in accumulation of fat in liver cells.
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