Gong Haixia, Ni Yuhui, Guo Xirong, Fei Li, Pan Xiaoqin, Guo Mei, Chen Ronghua
Department of Pediatrics, the second affiliated hospital of Nanjing Medical University, 121 Jiangjiayuan Road, Nanjing, 210011, People's Republic of China.
Eur J Endocrinol. 2004 Jun;150(6):885-92. doi: 10.1530/eje.0.1500885.
To investigate the relationship between resistin (a potential link between obesity and type 2 diabetes) and preadipocyte differentiation.
A rat resistin expression vector was transfected into 3T3-L1 preadipocytes and differentiation was compared between normal 3T3-L1 cells, rat resistin-transfected cells and non-transfected cells grown in conditioned medium taken from resistin-expressing cultures.
The rat resistin gene was inserted into the pDual GC and pEFGP-N2 expression vectors for examination of the effects of resistin overexpression in 3T3-L1 cells before and after differentiation was stimulated with 3-isobutyl-1-methyxanthine (MIX), insulin and dexamethasone (DEX). Smaller conserved fragments were inserted into short interference RNA (siRNA) expression vectors, for examination of the effect of targeted resistin inhibition on differentiation of resistin-overexpressing 3T3-L1 cells.
Prior to stimulation, the resistin-transfected 3T3-L1 cells contained many more small lipid droplets than did non-transfected 3T3-L1 cells. Following stimulation, differentiation in the resistin-transfected 3T3-L1 cells was dramatically promoted, especially in the early stages. Stimulation of differentiation was also observed in non-transfected 3T3-L1 cells grown in resistin protein-containing conditioned medium. The expression of adipocyte differentiation-associated markers such as CCAAT enhancer binding protein (C/EBPalpha), retinoid X receptor (RXRalpha) and lipoprotein lipase (LPL) was upregulated in resistin-overexpressing cells, whereas expression of preadipocyte factor-1 (Pref-1), an inhibitor of preadipocyte differentiation, was downregulated. In addition, expression of two of the three tested siRNAs inhibited the adipoconversion process, providing further evidence that resistin promotes the differentiation of preadipocytes to adipocytes.
Resistin can promote preadipocyte differentiation. Based on this, we propose that resistin may be an important candidate mediator of obesity-induced insulin resistance.
研究抵抗素（肥胖与2型糖尿病之间的潜在联系）与前脂肪细胞分化之间的关系。
将大鼠抵抗素表达载体转染至3T3-L1前脂肪细胞，并比较正常3T3-L1细胞、转染大鼠抵抗素的细胞以及在取自表达抵抗素培养物的条件培养基中生长的未转染细胞之间的分化情况。
将大鼠抵抗素基因插入pDual GC和pEFGP-N2表达载体，以检测在用3-异丁基-1-甲基黄嘌呤（MIX）、胰岛素和地塞米松（DEX）刺激分化前后，抵抗素过表达对3T3-L1细胞的影响。将较小的保守片段插入小干扰RNA（siRNA）表达载体，以检测靶向抑制抵抗素对过表达抵抗素的3T3-L1细胞分化的影响。
在刺激前，转染抵抗素的3T3-L1细胞比未转染的3T3-L1细胞含有更多的小脂滴。刺激后，转染抵抗素的3T3-L1细胞的分化显著促进，尤其是在早期阶段。在含有抵抗素蛋白的条件培养基中生长的未转染3T3-L1细胞中也观察到分化的刺激。在过表达抵抗素的细胞中，脂肪细胞分化相关标志物如CCAAT增强子结合蛋白（C/EBPα）、视黄酸X受体（RXRα）和脂蛋白脂肪酶（LPL）的表达上调，而前脂肪细胞分化抑制剂前脂肪细胞因子-1（Pref-1）的表达下调。此外，三种测试的siRNA中的两种的表达抑制了脂肪转化过程，进一步证明抵抗素促进前脂肪细胞向脂肪细胞的分化。
抵抗素可促进前脂肪细胞分化。基于此，我们提出抵抗素可能是肥胖诱导的胰岛素抵抗的重要候选介质。
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