Timirci-Kahraman Ozlem, Yilmaz Umit, Yilmaz Nesibe, Cevik Aydin, Horozoglu Cem, Celik Faruk, Gokce Muhammed Oguz, Ergen Arzu, Melekoglu Abdullah, Zeybek Umit
Department of Molecular Medicine, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey.
Department of Experimental Animal Biology and Biomedical Application Techniques, Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey.
In Vivo. 2018 Jul-Aug;32(4):813-817. doi: 10.21873/invivo.11312.
BACKGROUND/AIM: Adipocyte gene expression is altered in obese individuals through multiple metabolic and biochemical pathways. In this study, we aimed to examine the expression of resistin (Retn), amylin (Iapp), and dopamine receptor domain 5 (Drd5) genes previously suggested to contribute to the pathogenesis of obesity, albeit controversially. We also aimed to determine the effects on short and long-term mRNA levels of these genes in obese mice, induced with high-fat diet (HFD).
Two obesity models were created in our study: group T1 (20 mice) was fed with HFD (60% fat) for 3 months, and group T2 (20 mice) was fed with HFD (60% fat) for 6 months. The control group T0 (20 mice) was fed with a diet of 10% kcal fat supplement for 6 months. At the end of the experiment, their adipose tissues were dissected surgically. Tissue samples of each group were pooled for RNA isolation, cDNA synthesis was carried out and the mRNA levels were examined by quantitative real-time polymerase chain reaction. Serum resistin levels were measured using multiplex bead (luminex) technology for validation.
In T2 mice, the mRNA expression of Retn showed a moderate up-regulation (fold change=8.32; p=0.0019) in the adipose tissues. Iapp expression was also significantly up-regulated (fold change=9.78; p=0.012). Moreover, a 6.36-fold up-regulation for Drd5 was observed in the adipose tissues of T2 mice (p<0.001). At the same time, serum levels of resistin were found to be high in T1 and T2 mice compared to the control group (p<0.001 and p=0.024, respectively).
Our study demonstrated that the mRNA levels of the genetic markers considered to play a role in adipogenesis were different in short- and long-term obesity models formed in C57BL/6J mice using HFD.
背景/目的：肥胖个体的脂肪细胞基因表达通过多种代谢和生化途径发生改变。在本研究中，我们旨在检测抵抗素（Retn）、胰淀素（Iapp）和多巴胺受体D5（Drd5）基因的表达，这些基因此前虽存在争议，但被认为与肥胖发病机制有关。我们还旨在确定高脂饮食（HFD）诱导的肥胖小鼠中这些基因的短期和长期mRNA水平的变化。
我们在研究中创建了两种肥胖模型：T1组（20只小鼠）喂食高脂饮食（60%脂肪）3个月，T2组（20只小鼠）喂食高脂饮食（60%脂肪）6个月。对照组T0（20只小鼠）喂食含10%千卡脂肪补充剂的饮食6个月。实验结束时，手术解剖它们的脂肪组织。将每组的组织样本合并用于RNA分离，进行cDNA合成，并通过定量实时聚合酶链反应检测mRNA水平。使用多重珠（Luminex）技术测量血清抵抗素水平以进行验证。
在T2组小鼠中，Retn的mRNA表达在脂肪组织中呈中度上调（倍数变化=8.32；p=0.0019）。Iapp表达也显著上调（倍数变化=9.78；p=0.012）。此外，在T2组小鼠的脂肪组织中观察到Drd5上调了6.36倍（p<0.001）。同时，与对照组相比，T1组和T2组小鼠的血清抵抗素水平较高（分别为p<0.001和p=0.024）。
我们的研究表明，在使用高脂饮食在C57BL/6J小鼠中形成的短期和长期肥胖模型中，被认为在脂肪生成中起作用的遗传标记的mRNA水平有所不同。
