基础医学与临床 ›› 2012, Vol. 32 ›› Issue (3): 241-244.

• 研究论文 •    下一篇

Reg3α基因影响胰岛内皮细胞分泌胰岛素

滕兆刚1,范恒2,王立斌3,李玉奎1,魏军1   

  1. 1. 宁夏医科大学
    2. 宁夏医科大学总医院
    3. 宁夏医科大学附属医院
  • 收稿日期:2011-09-27 修回日期:2012-01-05 出版日期:2012-03-05 发布日期:2012-02-27
  • 通讯作者: 魏军 E-mail:lydiajunwei@hotmail.com.cn
  • 基金资助:
    胰岛β细胞体内再生过程的调节基因及其功能的研究

Effects of Reg3α gene in insulin secretion in pancreatic endothelial cells

  • Received:2011-09-27 Revised:2012-01-05 Online:2012-03-05 Published:2012-02-27

摘要: 目的 构建小鼠胰腺损伤模型,利用全基因组芯片技术筛选出高表达基因胰岛再生衍生因子3α(regenerating islet-derived 3 alpha,Reg3α),然后通过胰岛内皮细胞(MS1)过表达Reg3α,探讨小鼠Reg3α对胰腺再生中的重要作用。 方法 建立小鼠部分(90%)胰腺切除模型,分别于术前、术后12hr、24hr检测血糖, 术后48hr收集小鼠剩余胰腺组织,经全基因组芯片分析、q-PCR验证Reg3α高表达。构建Reg3α过表达质粒,转染MS1细胞系,48hr后ELISA检测细胞上清培养液胰岛素分泌量的变化,并通过q-PCR检测转染后MS1中Pdx1、Insulin2 mRNA表达水平。 结果 比较未转染组(untransfected group)与转染空载体组(PcDNA3.1-transfected group),MS1上清培养液胰岛素分泌量没有明显变化(14.63±6.88,P>0.05);比较转染Reg3α过表达质粒组与未转染组,上清培养液胰岛素分泌量明显升高(193.43±7.09, P<0.01);比较转染Reg3α过表达质粒组与转染空载体组,胰岛素分泌量有明显增加(208.07±5.52, P<0.01)。与未转染组、转染空载体组比较,转染Reg3α过表达质粒后MS1中的Pdx1、Insulin2 mRNA表达水平明显升高(P<0.01) 结论 胰岛内皮细胞中过表达Reg3α可使Pdx1、Insulin2 mRNA表达水平升高,增加胰岛素的分泌。Reg3α促进内皮细胞表达和分泌胰岛素可能具有对胰岛β细胞功能代偿作用。

关键词: Reg3α, 胰岛内皮细胞, 胰岛素, 转染

Abstract: Objective By establishing mouse model of pancreatic injury and regeneration and employing whole genome gene chip technology, to identify regenerating islet-derived 3 alpha(Reg3α)gene whose expression was elevated in pancreatic regenerating models, hence further investigate the regulatory role of Reg3α in pancreatic regeneration by over expression of Reg3α gene in pancreatic endothelial cells (MS1). Methods Mouse model of partial (90%) pancreas removal was established by panceatectomy. Blood glucose levels were tested before and 12hr, 24hr after pancreatectomy, respectively. The remaining pancreas tissues were collected 48h post pancreatectomy. Elevated expression of Reg3α gene was identified by gene chip analysis and verified by q-PCR. The gene was then cloned in an expression plasmid and expressed in MS1 cells in culture. Insulin levels in culture media of the transfected cells were detected by ELISA 48h after cell transfection. The mRNA levels of Pdx1 and Insulin2 genes in transfected cells were tested by q-PCR. Results Comparing untransfected group with vector (PcDNA3.1)-transfected group,no significant change of insulin levels in MS1 culture media was observed(14.63±6.88,P>0.05);when comparing Reg3α over expression group with untransfected and vector-transfected groups, significant increases in insulin levels were observed, 193.43±7.09 (P<0.01) over untransfected group, and 208.07±5.52 ( P<0.01) over vector-transfected group, respectively. Again, comparing with untransfected and vector-transfected groups, Reg3α over expression group expressed significantly higher mRNA levels of Pdx1 and Insulin2 genes(P<0.01). Conclusion Over expression of Reg3α gene in MS1 cells can elevate the expression of Pdx1 and Insulin2 in mRNA levels, and increase insulin secretion. Reg3α promote endothelial cell expression and secretion of insulin ,which may have a compensatory role in β-cell function.

Key words: Reg3α, MS1 Cells, Insulin, Transfection

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