MicroRNAs Expressed in Visceral Adipose Tissue from Mice Fed High-Fat Diets Supplemented with Eicosapentaenoic Acid (EPA)

The goal of this subproject is to identify microRNAs (miRNAs) expressed in visceral (VIS) adipose tissue (gonadal fat from male C57BL/6 mice) and regulated by eicosapentaenoic acid (EPA). This will provide insight into microRNAs regulated by EPA and their potential role in obesity-associated inflammation. We performed small RNA-sequencing of white (VIS) adipose tissue from high-fat diet (45% kcal from fat) supplemented with EPA (45% Kcal from fat, 6.75% EPA). Using the Gunaratne Next-Generation pipeline (published in Creighton et al. 2009), miRNA expression profiles were identified. Counts of each unique read were normalized to total usable reads, and had 40 counts added. We mapped about 13.8 million sequence reads per sample to the Mus musculus genome (build mm 10). We are specifically interested in those miRNAs expressed in VIS fat from EPA-fed mice compared to HF-fed mice. Overall design: Method: We performed small RNA-sequencing of VIS fat from mice fed high-fat diet supplemented with EPA (45% Kcal from fat, 6.75% EPA). Results: Using the Gunaratne Next-Generation pipeline (published in Creighton et al. 2009), miRNA expression profiles were identified. Counts of each unique read were normalized to total usable reads, and had 40 counts added. We mapped about 13.8 million sequence reads per sample to the Mus musculus genome (build mm 10). Conclusion: Specific miRNAs are expressed in white adipose tissue from mice fed EPA-supplemented HF diets.

本子课题的研究目标为鉴定在内脏脂肪组织（visceral adipose tissue, VIS，即雄性C57BL/6小鼠的性腺脂肪）中表达、且受二十碳五烯酸（eicosapentaenoic acid, EPA）调控的微小RNA（microRNAs, miRNAs）。本研究旨在阐明受EPA调控的miRNA及其在肥胖相关炎症中的潜在作用。我们对补充了EPA的高脂饮食（high-fat diet, HF，脂肪供能占总热量45%，其中EPA占6.75%）饲喂的小鼠的白色内脏脂肪组织实施了小RNA测序（small RNA-sequencing）。采用Gunaratne二代测序分析流程（Gunaratne Next-Generation pipeline，发表于Creighton等人2009年的研究），我们鉴定得到miRNA的表达谱。对每个唯一读段的计数以总可用读段数为基准进行归一化处理，并添加40个计数以作校正。我们将每个样本约1380万条测序读段比对至小家鼠（Mus musculus）基因组（版本mm10）。本研究重点关注EPA饲喂组小鼠与高脂饲喂组小鼠内脏脂肪中差异表达的miRNA。

总体实验设计：
实验方法：我们对补充了EPA的高脂饮食（脂肪供能占总热量45%，其中EPA占6.75%）饲喂的小鼠的内脏脂肪组织实施小RNA测序。
实验结果：采用Gunaratne二代测序分析流程（发表于Creighton等人2009年的研究），鉴定得到miRNA表达谱；对每个唯一读段的计数以总可用读段数为基准进行归一化，并添加40个计数以作校正；我们将每个样本约1380万条测序读段比对至小家鼠基因组（版本mm10）。
研究结论：在补充EPA的高脂饮食饲喂小鼠的白色脂肪组织中，存在特异性表达的miRNA。