Circulating LncRNAs Analysis in Patients with Type 2 Diabetes Reveals Novel Genes Influencing Glucose Metabolism and Islet β-Cell Function
Circulating LncRNAs Analysis in Patients with Type 2 Diabetes Reveals Novel Genes Influencing Glucose Metabolism and Islet β-Cell Function
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2 型糖尿病患者的循环 LncRNA 分析揭示了影响葡萄糖代谢和胰岛 β 细胞功能的新基因
DOI：
10.1159/000488434
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Sun, Jia
中科院分区：
文献类型：
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作者：
Ruan, Yuting;Lin, Nie;Sun, Jia
背景/目标：胰岛是重要的内分泌器官，分泌胰岛素调节糖代谢，维持血糖稳定。长链非编码RNA（lncRNA）参与多种生物学功能，并在包括2型糖尿病（T2 D）在内的许多疾病中发挥关键作用。本研究的目的是确定lncRNA-p3134是否与胰腺β细胞中的葡萄糖代谢和胰岛素信号传导相关。方法：应用LncRNA微阵列技术检测T2 D患者外周血中差异表达的lncRNA。采用RT-PCR方法检测30对糖尿病和非糖尿病患者lncRNA-p3134的表达。分析lncRNA-p3134与T2 D患者临床数据的相关性。通过腺病毒介导的技术在Min 6细胞和db/db小鼠中过表达LncRNA-p3134。采用CCK-8、TUNEL、Western blot、葡萄糖刺激的胰岛素分泌（GSIS）、ELISA和免疫组化等方法检测lncRNA-p3134在体外和体内对细胞增殖、凋亡和胰岛素分泌的影响。结果如下：糖尿病患者的lncRNA-p3134循环水平高于非糖尿病对照组，并与空腹血糖和HOMA-β水平相关。lncRNA-p3134在血清外泌体中上升了4倍，但在无外泌体的样品中几乎没有变化。葡萄糖在Min 6细胞和分离的小鼠胰岛细胞中动态调节lncRNA-p3134的分泌。LncRNA-p3134通过促进β细胞中的关键调节因子（Pdx-1、MafA、GLUT 2和Tcf 712）来正向调节GSIS。此外，lncRNA-p3134的过表达导致细胞凋亡率降低，并部分逆转了葡萄糖毒性对Min 6细胞GSIS功能的影响。db/db小鼠中胰岛素合成和分泌的恢复以及通过上调lncRNA-p3134而增加的胰岛素阳性细胞面积证实了lncRNA-p3134对保护β细胞功能的代偿作用。此外，还证实了lncRNA-p3134通过正性调节PI 3 K/Akt/mTOR信号传导对GSIS的保护作用。用其特异性抑制剂阻断PI 3 K/AKT信号后，过表达的lncRNA-p3134对胰岛素分泌的影响明显减弱。结论：总之，本研究的结果为胰腺β细胞中lncRNA-p3134的调控提供了新的见解，并为葡萄糖稳态的新机制提供了更好的理解。(C)2018作者（S）由S发布。Karger AG，巴塞尔
Background/Aims: The islet is an important endocrine organ to secrete insulin to regulate the metabolism of glucose and maintain the stability of blood glucose. Long noncoding RNAs (lncRNAs) are involved in a variety of biological functions and play key roles in many diseases, including type 2 diabetes (T2D). The aim of this study was to determine whether lncRNA-p3134 is associated with glucose metabolism and insulin signaling in pancreatic beta cells. Methods: LncRNA microarray technology was used to identify the differentially expressed circulating lncRNAs in T2D patients. RT-PCR analyses were performed to determine the expression of lncRNA-p3134 in 30 pairs of diabetic and non-diabetic patients. The correlation of lncRNA-p3134 to clinical data from T2D patients was analyzed. LncRNA-p3134 was overexpressed in Min6 cells and db/db mice by adenovirus-mediated technology. CCK-8, TUNEL, Western blot, glucose-stimulated insulin secretion (GSIS), ELISAs and immunochemistry were performed to determine the effect of lncRNA-p3134 on proliferation, apoptosis and insulin secretion both in vitro and vivo. Results: The circulating level of lncRNA-p3134 was higher in diabetic patients than in non-diabetic controls and was correlated with fasting blood glucose and HOMA-beta levels. The lncRNA-p3134 had risen by 4 times in serum exosomes but nearly unchanged in exosome-free samples. The secretion of lncRNA-p3134 was dynamically modulated by glucose in both Min6 cells and isolated mouse islet cells. LncRNA-p3134 positively regulate GSIS through promoting of key regulators (Pdx-1, MafA, GLUT2 and Tcf7l2) in beta cells. In addition, the overexpression of lncRNA-p3134 resulted in a decreased apoptosis ratio and partially reversed the glucotoxicity effects on GSIS function in Min6 cells. The restoration of insulin synthesis and secretion the increase of the insulin positive cells areas by upregulation of lncRNA-p3134 in db/db mice confirmed the compensatory role of lncRNA-p3134 to preserve beta-cell function. Furthermore, a protective effect of lncRNA-p3134 on GSIS by positive modulation of PI3K/Akt/mTOR signaling was also confirmed. After blocking the PI3K/AKT signals with their specific inhibitor, the effect of overexpressed lncRNA-p3134 on insulin secretion was obviously attenuated. Conclusion: Taken together, the results of this study provide new insights into lncRNA-p3134 regulation in pancreatic beta cells and provide a better understanding of novel mechanism of glucose homeostasis. (C) 2018 The Author(s) Published by S. Karger AG, Basel