LncRNA PLUTO促进胰岛素转录和分泌的作用及机制研究

张方方; 刘悦; 金亮

doi:10.11665/j.issn.1000-5048.20190414

LncRNA PLUTO促进胰岛素转录和分泌的作用及机制研究

中国药科大学生命科学与技术学院

基金项目: 国家自然科学基金资助项目(No.81570696)

计量
- 文章访问数: 443
- HTML全文浏览量: 0
- PDF下载量: 833
出版历程
- 刊出日期: 2019-08-24

Mechanism of lncRNA-PLUTO on promoting insulin biosynthesis and secretion

摘要

摘要: 探讨长链非编码RNA PLUTO对胰岛β细胞合成及分泌胰岛素作用的影响及机制。利用酶法分离提取db/db、ob/ob及HFD小鼠胰岛细胞，检测PLUTO在不同肥胖模型小鼠胰岛细胞中的表达量；利用糖脂毒性刺激胰岛原代细胞和Min6细胞，检测PLUTO的表达量，阐明肥胖因素诱导PLUTO下调的原因。通过RT-qPCR和ELISA方法分析PLUTO对胰岛素合成及分泌的影响；采用Western blot及功能回复实验阐明PLUTO调控胰岛β细胞的作用机制。结果表明:与正常小鼠相比，PLUTO在肥胖小鼠模型中表达量显著降低；在胰岛原代细胞和Min6细胞中过表达PLUTO促进胰岛合成和分泌；Western blot实验证明PLUTO通过上调相邻基因Pdx1的转录和翻译，促进胰岛素合成及分泌。
- LncRNA-PLUTO /
- 胰岛素合成 /
- 胰岛素分泌 /
- Pdx1 /
- 肥胖
Abstract: The aim of this study is to investigate the effect of long non-coding RNA PLUTO on β cell function and its underlying mechanism. Islet cells of db/db, ob/ob and HFD mice were isolated and extracted by enzymatic method to detect the expression of PLUTO in islet cells of different obesity model mice. Glucolipid toxicity was used to stimulate islet primary cells and Min6 cells, and the expression of PLUTO was detected to elucidate the reasons of PLUTO down-regulation induced by obesity factors. RT-qPCR and ELISA were performed to analyze the function of PLUTO on β cells; Western blot and rescue experiments were carried out to elucidate the regulatory mechanism of β cells by PLUTO. The results showed that the expression levels of PLUTO were significantly decreased in obesity model mice compared with control mice; over-expression PLUTO in primary islets and Min6 cells could improve insulin biosynthesis and secretion; Western blot analysis showed that PLUTO promoted insulin secretion and synthesis by up-regulating the transcription and translation of Pdx1, which is the adjacent gene of PLUTO.
- LncRNA PLUTO /
- insulin synthesis /
- insulin secretion /
- Pdx1 /
- obesity

HTML全文

参考文献(30)

[1]	Unger RH.Lipotoxicity in the pathogenesis of obesity-dependent NIDDM.Genetic and clinical implications[J].Diabetes,1995,44(8):863-870.
[2]	Zimmet P,Alberti KG,Shaw J.Global and societal implications of the diabetes epidemic[J].Nature,2001,414(6865):782-787.
[3]	Newsholme P,Keane D,Welters HJ,et al.Life and death decisions of the pancreatic beta-cell:the role of fatty acids[J].Clin Sci,2007,112(1):27-42.
[4]	Prentki M,Nolan CJ.Islet beta cell failure in type 2 diabetes[J].J Clin Invest,2006,116(7):1802-1812.
[5]	Saxena R,Voight BF,Lyssenko V,et al.Genome-wide association analysis identifies loci for type 2 diabetes and triglyceride levels[J].Science,2007,316(5829):1331-1336.
[6]	Scott LJ,Mohlke KL,Bonnycastle LL,et al.A genome-wide association study of type 2 diabetes in Finns detects multiple susceptibility variants[J].Science,2007,316(5829):1341-1345.
[7]	Yang F,Yi F,Zheng ZG,et al.Characterization of a carcinogenesis-associated long non-coding RNA[J].RNA Biol,2012,9(1):110-116.
[8]	Yu DM,Guo W,Lei W,et al.Advances of lncRNA in immune cells and autoimmune diseases [J].J China Pharm Univ(中国药科大学学报),2017,48(3):371-376.
[9]	Ponting CP,Oliver PL,Reik W.Evolution and functions of long noncoding RNAs[J].Cell,2009,136(4):629-641.
[10]	Széll M,Danis J,Bata-Csörgó Z,et al.PRINS,a primate-specific long non-coding RNA,plays a role in the keratinocyte stress response and psoriasis pathogenesis[J].Pflugers Arch,2016,468(6):935-943.
[11]	Yang YJ,Cai Y,Wu GZ,et al.Plasma long non-coding RNA,CoroMarker,a novel biomarker for diagnosis of coronary artery disease[J].Clin Sci,2015,129(8):675-685.
[12]	Kaur S,Mirza AH,Brorsson CA,et al.The genetic and regulatory architecture of ERBB3-type 1 diabetes susceptibility locus[J].Mol Cell Endocrinol,2016,419:83-91.
[13]	Mirza AH,Kaur S,Pociot F.Long non-coding RNAs as novel players in β cell function and type 1 diabetes[J].Hum Genomics,2017,11(1):17.
[14]	Wang M,Wang SY,Yao D,et al.A novel long non-coding RNA CYP4B1-PS1-001 regulates proliferation and fibrosis in diabetic nephropathy[J].Mol Cell Endocrinol,2016,426:136-145.
[15]	Benner C,van der Meulen T,Cacéres E,et al.The transcriptional landscape of mouse beta cells compared to human beta cells reveals notable species differences in long non-coding RNA and protein-coding gene expression[J].BMC Genomics,2014,15:620.
[16]	Ku GM,Kim H,Vaughn IW,et al.Research resource:RNA-Seq reveals unique features of the pancreatic β-cell transcriptome[J].Mol Endocrinol,2012,26(10):1783-1792.
[17]	Morán I,Akerman I,van de Bunt M,et al.Human β cell transcriptome analysis uncovers lncRNAs that are tissue-specific,dynamically regulated,and abnormally expressed in type 2 diabetes[J].Cell Metab,2012,16(4):435-448.
[18]	Arnes L,Akerman I,Balderes DA,et al.Βlinc1 encodes a long noncoding RNA that regulates islet β-cell formation and function[J].Genes Dev,2016,30(5):502-507.
[19]	You LH,Wang N,Yin DD,et al.Downregulation of long noncoding RNA meg3 affects insulin synthesis and secretion in mouse pancreatic beta cells[J].J Cell Physiol,2016,231(4):852-862.
[20]	Kallen AN, Zhou XB, Xu J, et al. The imprinted H19 lncRNA antagonizes let-7 microRNAs[J].Mol Cell,2013,52(1):101-112.
[21]	Geisler S,Coller J.RNA in unexpected places:long non-coding RNA functions in diverse cellular contexts[J].Nat Rev Mol Cell Biol,2013,14(11):699-712.
[22]	Pelechano V,Steinmetz LM.Gene regulation by antisense transcription[J].Nat Rev Genet,2013,14(12):880-893.
[23]	Tsai MC,Manor O,Wan Y,et al.Long noncoding RNA as modular scaffold of histone modification complexes[J].Science,2010,329(5992):689-693.
[24]	Fatica A,Bozzoni I.Long non-coding RNAs:new players in cell differentiation and development[J].Nat Rev Genet,2014,15(1):7-21.
[25]	Kopp F,Mendell JT.Functional classification and experimental dissection of long noncoding RNAs[J].Cell,2018,172(3):393-407.
[26]	Derrien T,Johnson R,Bussotti G,et al.The GENCODE v7 catalog of human long noncoding RNAs:analysis of their gene structure,evolution,and expression[J].Genome Res,2012,22(9):1775-1789.
[27]	Lovis P, Roggli E, Laybutt DR, et al. Alterations in microRNA expression contribute to fatty acid-induced pancreatic beta-cell dysfunction[J].Diabetes,2008,57(10):2728-2736.
[28]	Heilmann K,Toth R,Bossmann C,et al.Genome-wide screen for differentially methylated long noncoding RNAs identifies Esrp2 and lncRNA Esrp2-as regulated by enhancer DNA methylation with prognostic relevance for human breast cancer[J].Oncogene,2017,36(46):6446-6461.
[29]	Spaeth JM,Walker EM,Stein R.Impact of Pdx1-associated chromatin modifiers on islet β-cells[J].Diabetes Obes Metab,2016,18(Suppl 1):123-127.
[30]	Wang WP,Wang XF,Peng SY,et al.Transcriptional regulation of insulin gene [J].Chem Life,2005,25(2):103-105.