天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2014年
4期
289-292
,共4页
张君%荣曦%吴宇娟%刘红
張君%榮晞%吳宇娟%劉紅
장군%영희%오우연%류홍
胰岛素原%胰岛素分泌细胞%硝苯地平%胰岛素受体底物1%酪氨酸磷酸化%HIT-T15细胞
胰島素原%胰島素分泌細胞%硝苯地平%胰島素受體底物1%酪氨痠燐痠化%HIT-T15細胞
이도소원%이도소분비세포%초분지평%이도소수체저물1%락안산린산화%HIT-T15세포
proinsulin%insulin-secreting cells%nifedipine%insulin receptor substrate1%tyrosine phosphorylation%HIT-T15 cells
目的：探讨短期外源性胰岛素刺激对HIT-T15胰岛β细胞胰岛素原（PI）基因表达的影响及机制。方法 HIT-T15仓鼠胰岛瘤细胞随机分为4组：含1.4 mmol/L葡萄糖完全培养基组作为空白对照(LG组)，为抑制内源性胰岛素释放干扰以低糖联合硝苯地平作为条件对照（LGC组），在LGC基础上分别加0.5 U/L或5 U/L的外源性胰岛素作为实验组(分别为LINS组和HINS组)，分别在刺激后0、30、60、90、120 min以荧光定量PCR检测各组PI mRNA，免疫组化检测各组细胞胰岛素受体底物1（IRS1）酪氨酸磷酸化水平。结果（1）LINS组、HINS组细胞PI mRNA表达均上调，在刺激后60 min PI mRNA表达量达高峰。（2）实验组在刺激后30 min IRS1酪氨酸磷酸化水平较对照组明显增高，高、低浓度胰岛素组细胞酪氨酸磷酸化达高峰时间不同。（3）LG组和LGC组间PI mRNA表达及IRS1酪氨酸磷酸化水平无明显差异。结论短期外源性胰岛素能上调胰岛β细胞PI基因的表达，且高浓度的胰岛素较低浓度胰岛素作用强。PI表达调控与IRS1信号传导有关。
目的：探討短期外源性胰島素刺激對HIT-T15胰島β細胞胰島素原（PI）基因錶達的影響及機製。方法 HIT-T15倉鼠胰島瘤細胞隨機分為4組：含1.4 mmol/L葡萄糖完全培養基組作為空白對照(LG組)，為抑製內源性胰島素釋放榦擾以低糖聯閤硝苯地平作為條件對照（LGC組），在LGC基礎上分彆加0.5 U/L或5 U/L的外源性胰島素作為實驗組(分彆為LINS組和HINS組)，分彆在刺激後0、30、60、90、120 min以熒光定量PCR檢測各組PI mRNA，免疫組化檢測各組細胞胰島素受體底物1（IRS1）酪氨痠燐痠化水平。結果（1）LINS組、HINS組細胞PI mRNA錶達均上調，在刺激後60 min PI mRNA錶達量達高峰。（2）實驗組在刺激後30 min IRS1酪氨痠燐痠化水平較對照組明顯增高，高、低濃度胰島素組細胞酪氨痠燐痠化達高峰時間不同。（3）LG組和LGC組間PI mRNA錶達及IRS1酪氨痠燐痠化水平無明顯差異。結論短期外源性胰島素能上調胰島β細胞PI基因的錶達，且高濃度的胰島素較低濃度胰島素作用彊。PI錶達調控與IRS1信號傳導有關。
목적：탐토단기외원성이도소자격대HIT-T15이도β세포이도소원（PI）기인표체적영향급궤제。방법 HIT-T15창서이도류세포수궤분위4조：함1.4 mmol/L포도당완전배양기조작위공백대조(LG조)，위억제내원성이도소석방간우이저당연합초분지평작위조건대조（LGC조），재LGC기출상분별가0.5 U/L혹5 U/L적외원성이도소작위실험조(분별위LINS조화HINS조)，분별재자격후0、30、60、90、120 min이형광정량PCR검측각조PI mRNA，면역조화검측각조세포이도소수체저물1（IRS1）락안산린산화수평。결과（1）LINS조、HINS조세포PI mRNA표체균상조，재자격후60 min PI mRNA표체량체고봉。（2）실험조재자격후30 min IRS1락안산린산화수평교대조조명현증고，고、저농도이도소조세포락안산린산화체고봉시간불동。（3）LG조화LGC조간PI mRNA표체급IRS1락안산린산화수평무명현차이。결론단기외원성이도소능상조이도β세포PI기인적표체，차고농도적이도소교저농도이도소작용강。PI표체조공여IRS1신호전도유관。
Objective To investigate the short term effect of insulin on proinsulin gene expression of HIT-T15 insu-linnoma cells(pancreatic isletβ-cell). Methods The HIT-T15 cells were randomly divided into four groups.Blank Con-trol Group (LG):complete medium contain 1.4 mmol/L glucose. Control group (LGC):co-cultured nifedipine with medium in order to restain endogenous insulin release. Experimental group (LINS or HINS) add 0.5 U/L insulin or 5 U/L insulin on top of LGC. After being stimulated for 0, 30, 60, 90, 120 mins, proinsulin (PI) mRNA level were assessed by semi-quantitative RT-PCR. Insulin receptor substrate1 (IRS1) tyrosine phosphorylation was detected by immunocytochemistry. Results (1) Expression of PI was up regulated by both LINS and HINS, and peak at 60 mins. (2) After stimulation for 30 mins, the level of IRS1 tyrosine phosphorylation in the experimental group was significantly higher than control group, and the peak time be-tween LINS and HINS was different. (3) Between group of LG and LGC, the expression of PI mRNA and IRS1 tyrosine phos-phorylation show no difference. Conclusion Short term exogenous insulin stimulation can promote expression of proinsulin genes,which is concentration dependent. The expression and regulation of PI were related with IRS1 signal transduction.
