基础医学与临床
基礎醫學與臨床
기출의학여림상
BASIC MEDICAL SCIENCES AND CLINICS
2010年
4期
389-393
,共5页
林梅%张木勋%刘永健%张建华%余毅恺%帅红霞
林梅%張木勛%劉永健%張建華%餘毅愷%帥紅霞
림매%장목훈%류영건%장건화%여의개%수홍하
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
RNA榦擾%11β-羥類固醇脫氫酶1型%NIT-1細胞%葡萄糖刺激胰島素分泌
RNA간우%11β-간류고순탈경매1형%NIT-1세포%포도당자격이도소분비
RNA interference%11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1)%NIT-1 cell%glucose-stimulated insulin secre-tion (GSIS)
目的 观察RNA干扰阻断胰岛β细胞系NIT-1中11β-羟类固醇脱氧酶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-HSD1 mRNA和蛋白表达.oligo886重组质粒转染25 mmol/L葡萄糖培养摹中的NIT-1细胞,测GSIS.结果 转染oligo886和oligo866质粒的NIT-1细胞11β-HSD1 mRNA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.
目的 觀察RNA榦擾阻斷胰島β細胞繫NIT-1中11β-羥類固醇脫氧酶1型(11β-HSD1)錶達對細胞葡萄糖刺激胰島素分泌(GSIS)的影響.方法 pSuper質粒為載體,構建針對11β-HSD1基因的質粒oligo886、oligo866和亂序對照質粒oligoScr886,轉染NIT-1細胞,RT-PCR和蛋白印跡法檢測11β-HSD1 mRNA和蛋白錶達.oligo886重組質粒轉染25 mmol/L葡萄糖培養摹中的NIT-1細胞,測GSIS.結果 轉染oligo886和oligo866質粒的NIT-1細胞11β-HSD1 mRNA分彆下降78.1%±2.9%和51.7%±2.7%,11β-HSD1蛋白的水平分彆下降82.2%±2.1%和56.5%±2.0%.oligo866轉染25 mmoL/L葡萄糖培養基中的NIT-1細胞後,GSIS較對照組明顯升高(P<0.01).結論 11β-HSD1基因沉默能改善NIT-1細胞的葡萄糖刺激胰島素分泌能力,提示胰島β細胞NIT-1通過11β-HSD1調節糖皮質代謝,參與葡萄糖刺激胰島素分泌.
목적 관찰RNA간우조단이도β세포계NIT-1중11β-간류고순탈양매1형(11β-HSD1)표체대세포포도당자격이도소분비(GSIS)적영향.방법 pSuper질립위재체,구건침대11β-HSD1기인적질립oligo886、oligo866화란서대조질립oligoScr886,전염NIT-1세포,RT-PCR화단백인적법검측11β-HSD1 mRNA화단백표체.oligo886중조질립전염25 mmol/L포도당배양모중적NIT-1세포,측GSIS.결과 전염oligo886화oligo866질립적NIT-1세포11β-HSD1 mRNA분별하강78.1%±2.9%화51.7%±2.7%,11β-HSD1단백적수평분별하강82.2%±2.1%화56.5%±2.0%.oligo866전염25 mmoL/L포도당배양기중적NIT-1세포후,GSIS교대조조명현승고(P<0.01).결론 11β-HSD1기인침묵능개선NIT-1세포적포도당자격이도소분비능력,제시이도β세포NIT-1통과11β-HSD1조절당피질대사,삼여포도당자격이도소분비.
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.
