摘要
目的探讨血管内皮生长因子(vascular endothelial growth factor,VEGF)对大鼠胰岛瘤细胞(INS-1)增殖、凋亡、胰岛素分泌以及相关基因表达的影响。方法用不同浓度(0、40、80、160ng/mL)VEGF对大鼠INS-1细胞进行处理,采用CCK-8试剂盒检测INS-1细胞增殖,用AnnexinⅤ及碘化丙啶(PI)双染试剂检测细胞凋亡;INS-1细胞经VEGF处理后做标准葡萄糖刺激的胰岛素分泌实验,ELISA法检测胰岛素,实时定量PCR技术检测胰岛分泌过程中相关基因表达,Western blot检测VEGF对Insulin蛋白表达的影响。结果不同浓度VEGF对INS-1细胞作用24h、48h、72h,其细胞活性均无明显变化(P>0.05)。但当VEGF浓度为80ng/mL和160ng/mL时对细胞凋亡具有抑制作用(P<0.01)。高糖状态下当VEGF浓度为40ng/mL时,VEGF对胰岛素的分泌具有抑制作用。不同浓度VEGF作用于INS-1细胞后,磺酰脲类受体基因(sulfonylurea receptor,Sur)、内向整流性钾离子通道基因(inwardly rectifying potassium channel 6.2,Kir6.2)的表达随VEGF浓度增高呈下降趋势,葡萄糖激酶基因(glucokinase,GCK)的表达先降低后升高,葡萄糖转运蛋白基因2(glucose transporter 2,Glut2)表达呈先升高后降低趋势,Insulin蛋白的表达量随VEGF浓度增高呈逐渐下降趋势。结论 VEGF在高糖状态下对细胞凋亡和胰岛素分泌有抑制作用,为探索VEGF在糖代谢中的作用提供了新的线索。
Objective To investigate the effect of vascular endothelial growth factor（VEGF）on proliferation,apoptosis,insulin secretion and related gene expression in rat pancreatic islet cell（INS-1）.MethodsINS-1cells were treated with different concentrations of VEGF.CCK-8kit was used to detect the proliferation of INS-1cells and the cell apoptosis were evaluated by using AnnexinⅤ and propidium iodide（PI）double staining kit.INS-1cells were treated with VEGF and the standard glucose stimulated insulin secretion test with ELISA was conducted.The expression of related genes in pancreatic islets was detected by real-time quantitative PCR.The effect of VEGF on isulin protein expression was evaluated with Western blot.Results No significant changes（P〉0.05）in INS-1cells were observed after treated with different concentrations of VEGF at 24 h,48hand 72 h.But when VEGF concentration were 80ng/mL and 160ng/mL,an inhibitory effect on cell apoptosis were noticed（P〈0.01）.The addition of VEGF to the high-glucose media significantly reduced the release of insulin at the concentration of 40ng/mL.A decreasing trends of the expression level of sulfonylurea receptor gene（Sur）,inwardly rectifying potassium channel gene 6.2（Kir6.2）as well as the release of insulin were noticed as the increasing of VEGF concentrations.The expression of glucokinase gene（GCK）first decreased and then increased,but the expression of glucose transporter gene 2（Glut 2）were increased first and then decreased.Conclusion VEGF inhibited the secretion of insulin from INS-1cells in the high-glucose condition.Our study provides new clues to the function of VEGF on the glucose metabolism.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2016年第1期33-38,共6页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(No.82172684
No.81200640)资助
