Inhibition of 3T3-L1 adipocyte differentiation by 6-ethoxyzolamide:: repressed peroxisome proliferator-activated receptor γ mRNA and enhanced CCAAT/enhancer binding protein β mRNA levels
Inhibition of 3T3-L1 adipocyte differentiation by 6-ethoxyzolamide:: repressed peroxisome proliferator-activated receptor γ mRNA and enhanced CCAAT/enhancer binding protein β mRNA levels
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DOI：
10.1016/j.bcp.2003.12.039
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发表时间：
2004-05-01
影响因子：
5.8
通讯作者：
Tsuchida, S
中科院分区：
文献类型：
--
作者：
Takahata, T;Kumano, T;Tsuchida, S
应用细胞内脂肪储存定量图像分析技术，研究了碳酸酐酶（CA）抑制剂6-乙氧唑胺（ETZ）对小鼠前脂肪细胞系3 T3-L1分化的影响。对于脂肪细胞分化，融合后的细胞用过氧化物酶体增殖物激活受体γ（PPARgamma）的配体、曲格列酮（TRG）和地塞米松处理2天。分化的细胞在第4天显示出弱的脂肪染色，此后增加，与CAIII表达相关。ETZ在200 μ M浓度下处理10天，降低了储存脂肪的分化脂肪细胞的百分比和储存在单个细胞中的脂肪量。这些发现也得到了荧光激活细胞分选分析结果的支持。然而，尽管它们的脂肪染色较浅，但在ETZ处理的细胞中不表达CAIII。此外，ETZ延迟了细胞的克隆扩增，这是分化前的早期事件。北方印迹分析显示ETZ处理的细胞中CCAAT/增强子结合蛋白（C/EBP）β mRNA水平较高，而PPARgamma mRNA水平较低。因此，在这种情况下，C/EBP β mRNA的增加不会导致PPARgamma表达的增强。另一种CA抑制剂乙酰唑胺不抑制脂肪细胞分化，尽管该药物表现出与ETZ相似的CA活性抑制模式。这些结果表明ETZ对脂肪细胞分化的抑制作用不是由于CA活性的抑制，而是由于转录因子水平的改变。(C)2004年爱思唯尔公司All rights reserved.
The effects of 6-ethoxyzolamide (ETZ), a carbonic anhydrase (CA) inhibitor, on differentiation of the mouse preadipocyte cell line 3T3-L1 were examined by quantitative image analysis of intracellular fat storage. For adipocyte differentiation, postconfluent cells were treated with a ligand of the peroxisome proliferator-activated receptor gamma (PPARgamma), troglitazone (TRG), and dexamethasone for 2 days. Differentiated cells showed weak fat staining at day 4 which increased thereafter, correlating with CAIII expression. ETZ treatment for 10 days at a 200 muM concentration reduced both the percentage of differentiated adipocytes storing fat and the amount of fat stored in individual cells. These findings were also supported by the results of fluorescence activated cell sorting analysis. Despite their light fat staining, however, CAIII was not expressed in ETZ-treated cells. Furthermore, ETZ delayed the clonal expansion of cells, an early event preceding differentiation. Northern blot analysis revealed a high CCAAT/enhancer binding protein (C/EBP) beta mRNA level and low PPARgamma mRNA in ETZ-treated cells. Thus, increased C/EBPbeta mRNA did not lead to enhanced PPARgamma expression in this case. Another CA inhibitor acetazolamide did not inhibit adipocyte differentiation, although the drug exhibited a similar inhibition pattern for CA activity as ETZ. These results suggested that inhibitory effects of ETZ on adipocyte differentiation were not due to inhibition of CA activity but rather due to altered levels of the transcription factors. (C) 2004 Elsevier Inc. All rights reserved.