ABSTRACT
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.O1igo886 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% inolig886 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 olig 8 8 6 vector,the insulin secretion increased in NIT -1 cell.Conclusion 11β-HSD1 gene silencing may improve GSIS in NIT-1 cell 11β-HSD1 regulate local glucocorticoid metabolism in pan-creatic islet and affect the function of insulin secretion.
ABSTRACT
Overexpression of survivin may partly protect the NIT-1 cells(mouse insulin-secreting cells) from cytokine-induced apoptosis.In addition, NIT-1 cells transfected with survivin had an slightly improved response of insulin secretion to glucose stimulation.