基础医学与临床 ›› 2010, Vol. 30 ›› Issue (4): 389-393.

• 研究论文 • 上一篇    下一篇

11β-羟类固醇脱氢酶1型基因沉默对胰岛β细胞系NIT-1葡萄糖刺激胰岛素分泌的影响

林梅 张木勋 刘永健 张建华 余毅恺 帅红霞   

  1. 华中科技大学同济医学院普爱医院 华中科技大学同济医学院同济医院内分泌科
  • 收稿日期:2009-10-09 修回日期:2009-11-16 出版日期:2010-04-05 发布日期:2010-04-05
  • 通讯作者: 张木勋

Effect of 11β-hydroxysteroid dehydrogenase type 1 gene silencing on glucose-stimulated insulin secretion of pancreatic β cell line NIT-1

Mei LIN, Mu-xun ZHANG, Yong-jian LIU, Jian-hua ZHANG, Yi-kai YU, Hong-xia SHUAI   

  1. Puai Hospital, Tongji Medical College, Huazhong University of Science and Technology .Department of Endocrinology, Tongji Hospital,Tongji Medical College, Huazhong University of Science and Technology
  • Received:2009-10-09 Revised:2009-11-16 Online:2010-04-05 Published:2010-04-05
  • Contact: Mu-xun ZHANG,

摘要: 目的 观察RNA干扰阻断胰岛β细胞系NIT-1中11β-羟类固醇脱氢酶1型(11β-HSD1)表达对细胞葡萄糖刺激胰岛素分泌(GSIS)的影响。方法 pSuper质粒为载体,构建针对11β-HSD1基因的质粒oligo886、oligo866和乱序对照质粒oligoScr886,转染NIT-1细胞,RP-PCR和蛋白印迹法检测11β-HSD1mRNA和蛋白表达。oligo886重组质粒对25mmol/L葡萄糖培养基中NIT-1细胞进行转染,测GSIS。结果 转染oligo886和oligo866质粒NIT-1细胞11β-HSD1mRNA下降(78.1±2.9)%和(51.7±2.7)%,11β-HSD1蛋白的水平下降(82.2±2.1)%和(56.5±2.0)%。oligo866转染25mmol/L浓度葡萄糖培养基中的NIT-1细胞后,GSIS较对照组明显升高(P<0.01)。结论 11β-HSD1基因沉默能改善NIT-1细胞的葡萄糖刺激胰岛素分泌能力,提示胰岛β细胞NIT-1通过11β-HSD1调节糖皮质代谢,参与葡萄糖刺激胰岛素分泌。

关键词: RNA 干扰, 11β-羟类固醇脱氢酶1型, NIT-1细胞, 葡萄糖刺激胰岛素分泌

Abstract: Objective To investigate the effect of small interference RNA (siRNA) targeting 11β-hydroxysteroid dehydrogenase type 1 on the glucose-stimulated insulin secretion (GSIS) in pancreatic β cell line NIT-1 cell. Methods siRNA plasmid vectors specifically targeting 11β-HSD1 gene were constructed , named as olig886, oligo866 and scrabble control for oligo886, and tansfected into NIT-1 cells. The expression of 11β-HSD1 was detected by RT-PCR and Western blot. Oligo886 vector was transfected into the NIT-1 cells in 25mmol/L glucose concentrations medium .The insulin secretion level was measured in GSIS test. Results After treatment with 11β-HSD1 siRNA, the mRNA levels of 11β-HSD1 in NIT-1 cell were decreased by (78.1±2.9)% and (51.7±2.7)% in olig886 and oligo866 group. The protein of 11β-HSD1 were decreased by(82.2±2.1) % and (56.5±2.0)%. After transfected by olig886 vector, the insulin secretion increased in NIT-1 cell in 25mmol/L glucose concentration medium, contrasting with control group(P<0.01). Conclusion 11β-HSD1 gene silencing could improve GSIS in NIT-1 cell in . These suggested the 11β-HSD1 regulate local glucocorticoid metabolism in pancreatic islet, therefore, affect the function of insulin secretion.

Key words: RNA interference, 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), NIT-1 cell, glucose-stimulated insulin secretion (GSIS)