The aim of the present study was to test the influence of obesity and the presence of type 2 diabetes mellitus (T2DM) on the expression of ten housekeeping genes and of the 18S rRNA in a group of human adipose tissue samples from the omental and subcutaneous depot. Adipose tissue biopsies were obtained by laparoscopic surgery from lean and obese patients. After the extraction, mRNA levels in adipose tissue samples were quantified by real-time PCR using the commercial HUMAN ENDOGENOUS CONTROL PLATES. From the genes analyzed, 18S rRNA exhibited the most stable expression levels in both depots regardless of the pathophysiological conditions of obesity and obesity-associated T2DM. Contrarily, GAPD was the gene with the highest variation in its expression levels, being upregulated (8.0-fold) in the obese group and downregulated (3.5-fold) in obesity-associated T2DM. Our results show that 18S rRNA may be the most suitable gene for normalization in expression studies performed in human adipose tissue samples obtained from patients suffering from obesity and/or obesity-associated T2DM, whereas GAPD is less appropriate for comparison purposes under these circumstances.

译文

本研究的目的是测试肥胖和2型糖尿病 (T2DM) 的存在对来自网膜和皮下仓库的一组人类脂肪组织样品中十个管家基因和18S rRNA表达的影响。脂肪组织活检是通过腹腔镜手术从瘦和肥胖患者中获得的。提取后,使用商业人类内源性对照板通过实时PCR定量脂肪组织样品中的mRNA水平。从分析的基因来看,无论肥胖和肥胖相关的T2DM的病理生理状况如何,18S rRNA在两个仓库中均表现出最稳定的表达水平。相反,GAPD是其表达水平变化最高的基因,在肥胖组中上调 (8.0倍),在肥胖相关的T2DM中下调 (3.5倍)。我们的结果表明,在从肥胖和/或肥胖相关的T2DM患者获得的人脂肪组织样本中进行的表达研究中,18S rRNA可能是最适合标准化的基因,而在这些情况下,GAPD不太适合用于比较目的。

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