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  

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

作　　者：林梅[1] 张木勋[2] 刘永健[1] 张建华[2] 余毅恺[2] 帅红霞[2] 

机构地区：[1]华中科技大学同济医学院普爱医院内分泌科,湖北武汉430033 [2]华中科技大学同济医学院同济医院内分泌科,湖北武汉430030

出　　处：《基础医学与临床》2010年第4期389-393,共5页Basic and Clinical Medicine

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,RT-PCR和蛋白印迹法检测11β-HSD1mRNA和蛋白表达。oligo886重组质粒转染25 mmol/L葡萄糖培养基中的NIT-1细胞,测GSIS。结果转染oligo886和oligo866质粒的NIT-1细胞11β-HSD1mRNA分别下降78.1%±2.9%和51.7%±2.7%,11β-HSD1蛋白的水平分别下降82.2%±2.1%和56.5%±2.0%。oligo866转染25 mmol/L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。Objective To investigate the effect of small interference RNA（siRNA） targeting at 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 at 11β-HSD1 gene were constructed,named as olig886,oligo866 and scrabble control for oligo886,then 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 25 mmol/L glucose concentrations medium.The insulin secretion level was measured in GSIS test.Results After treatment with 11β-HSD1 siRNA,the mRNA level of 11β-HSD1 in NIT-1 cell was decreased by 78.1%±2.9% and 51.7%±2.7% in olig886 and oligo866 group respectively.The protein of 11β-HSD1 were decreased by 82.2%±2.1% and 56.5%±2.0% respectively.After transfected by olig886 vector,the insulin secretion increased in NIT-1 cell.Conclusion11β-HSD1 gene silencing may improve GSIS in NIT-1 cell 11β-HSD1 regulate local glucocorticoid metabolism in pancreatic islet and affect the function of insulin secretion.

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 

分 类 号：R587.1[医药卫生—内分泌]