Identification of transcription activators of resistin gene with type 2 diabetes susceptibility promoter SNP

抵抗素，由脂肪细胞分泌，引起啮齿类动物的胰岛素抵抗。在人类中，抵抗素的主要来源被认为是单核细胞，其病理生理相关性一直存在争议。我们之前报道了抵抗素基因启动子单核苷酸多态性（SNP） at-420的G/G基因型通过增强启动子活性增加2型糖尿病（T2DM）的易感性。我们还发现循环抵抗素与SNP-420的G等位基因数量相关，并与胰岛素抵抗相关。为了确定胰岛素增敏药物特异性影响抵抗素基因启动子的靶点，包括T2DM易感等位基因G - SNP-420，我们分析了抵抗素基因在THP-1人单核细胞中的表达机制。我们制作了荧光素酶报告基因，包括人类抵抗素基因在SNP-420上带有C或G的不同长度的5'侧区。我们发现5'侧区具有启动子活性。我们还分析了激素和药物对THP-1细胞抵抗素mRNA的影响。一些分子以剂量和时间依赖的方式影响抵抗素mRNA。然而，这些分子并不影响抵抗素启动子的活性。在THP-1细胞的细胞裂解液和培养液中均未检测到抵抗素蛋白。因此，通过分析THP-1细胞中抵抗素基因mRNA的表达可以最灵敏地评估其表达。在新鲜分离的人单核细胞中，抵抗素mRNA与其同期血清水平呈正相关，且在G/G基因型中最高。在T2DM患者中，晚期微血管病变患者血清抵抗素较高。这些发现表明，人类单核细胞中的抵抗素mRNA与SNP-420相关，而SNP-420与血清抵抗素相关，并且血清抵抗素在T2DM患者中较高，尤其是晚期微血管病变患者。因此，在单核细胞中发现降低抵抗素基因转录或mRNA的药物可以预防T2DM的发生及其并发症的进展。少

Resistin, secreted from adipocytes, causes insulin resistance in rodents. In humans, the main source of resistin is thought to be monocytes, and its pathophysiological relevance has been controversial. We previously reported that the G/G genotype of a resistin gene promoter single nucleotide polymorphism (SNP) at-420 increases-type 2 diabetes (T2DM) susceptibility by enhancing promoter activity. We also found that circulating resistin was associated with the G allele number of SNP-420, and correlated with insulin resistance. To identify targets of insulin sensitizing drugs specifically affecting the resistin gene promoter including T2DM susceptibility allele, G at SNP-420, we analyzed the mechanism of resistin gene expression in THP-1 human monocytes.. We made luciferase reporters including different length of 5' flanking region of the human resistin gene with either C or G at SNP-420. We found that the 5' flanking region had the promoter activities. We also analyzed effects of hormone … More s and drugs on resistin mRNA in THP-1 cells. Some molecules affected resistin mRNA in a dose and time dependent manner. However, these molecules did not affect resistin promoter activities. We did not detect resistin protein in cell lysate and cultured medium in THP-1 cells. Therefore, resistin gene expression could be most sensitively assessed by analyzing its mRNA in THP-1 cells. In freshly isolated human monocytes, resistin mRNA was positively correlated with its simultaneous serum levels, and highest in G/G genotype. In T2DM, serum resistin was higher in subjects with advanced microangiopathies. These findings suggest that resistin mRNA in human monocytes is associated with SNP-420, which is correlated with serum resistin, and that serum resistin is higher in T2DM, especially with advanced microangiopathies. Therefore, the identification of agents reducing resistin gene transcription or mRNA in monocytes could prevent the development of T2DM and the progression of its complications. Less