高浓度葡萄糖条件下罗格列酮对NIT-1细胞FOXO1、TSC2基因表达及细胞分泌功能的影响  

Effects of rosiglitazone on expressions of FOXO1 and TSC2 gene and cell secretory function of NIT-1 cells after treated with high concentration glucose

作　　者：乔伟[1] 刘丹[1] 孙情[1] 梁瑜祯[2] 冯乐平[1] 

机构地区：[1]桂林医学院生物技术学院实验教学中心,广西桂林541004 [2]广西医科大学第一附属临床医院代谢糖尿病中心,广西南宁530021

出　　处：《吉林大学学报（医学版）》2010年第2期252-257,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30860116);广西教育厅科研基金资助课题(院科字[2008]17号)

摘　　要：目的:研究罗格列酮在不同浓度葡萄糖条件下,对胰岛β细胞增殖凋亡与胰岛素分泌以及叉头转录因子-1(FOXO1)和结节性硬化症-2(TSC2)表达的影响。方法:将NIT-1细胞按每孔5×104个放置于24孔细胞培养板,培养48h后随机分为各处理组:5.6、7.8、11.1、16.7、22.2和27.6mmol.L-1葡萄糖组,继续培养24h后再分别施加1×10-5mol.L-1罗格列酮,分别于干预24和48h后取细胞培养上清液,采用放射免疫法检测胰岛素水平和免疫荧光法检测细胞增殖情况,RT-PCR半定量法检测FOXO1和TSC2mRNA表达水平。结果:①1×10-6～1×10-5mol.L-1罗格列酮可以分别在不同浓度葡萄糖培养条件下使胰岛NIT-1细胞增殖(P<0.05),且这种变化趋势随剂量的增加而增加(即1×10-5mol.L-1罗格列酮组>1×10-6mol.L-1罗格列酮组>1×10-7mol.L-1罗格列酮组)。1×10-5mol.L-1的罗格列酮干预后,可见细胞凋亡百分率增加趋势随着葡萄糖浓度不断升高;②在同一浓度罗格列酮作用下,当葡萄糖浓度为11.1mmol.L-1时,胰岛素分泌水平最高,高于其他各组(均P<0.05),随着葡萄糖浓度增加,胰岛素分泌量逐渐下降(11.1mmol.L-1葡萄糖组>16.7mmol.L-1葡萄糖组>22.5mmol.L-1葡萄糖组>27.6mmol.L-1葡萄糖组),而葡萄糖为5.6mmol.L-1时,胰岛素分泌量最低;③在1×10-5mol.L-1罗格列酮干预后,FOXO1和TSC-2mRNA的表达水平均较未干预组明显下降,且呈现出5.6mmol.L-1组<11.1mmol.L-1组<16.7mmol.L-1组<22.5mmol.L-1组<27.6mmol.L-1组的变化趋势,而且葡萄糖浓度>16.7mmol.L-1的各组均较前面小剂量葡萄糖组(≤11.1mmol.L-1各组)表达明显。结论:罗格列酮可以通过直接影响胰岛β细胞内FOXO1和TSC2表达促进胰岛β细胞的增殖及影响细胞胰岛素分泌功能,提示通过调控FOXO1和TSC2表达,可以直接影响胰岛β细胞的生物学功能,如分泌功能、细胞的增殖与凋亡以及改善胰岛素抵抗状况。Objective To study the effects of rosiglitazone on FOXO1 and TSC2 gene expressions,insulin secretory function,cell proliferation and apoptosis of pancreatic β cells under high concentration glucose condition.Methods The NIT-1 cells were put into plates（5×104 cells /well） and cultivated for 48 h,then they were randomly divided into treatment groups containing different concentrations of glucose as follows：5.6,7.8,11.1,16.7,22.2,and 27.6 mmol·L-1 groups.After cultivated for 24 h,they were intervented by 10-5 mmol·L-1 rosiglitazone for next 24 and 48 h,then the supernatant was collected.The insulin level was evaluated by radio-immunity technique,the cell proliferation and apoptosis were detected by immunofluorescence staining and MTT assay respectivly.The expressions of FOXO1 and TSC2 mRNA were detected by semi-quantitative RT-PCR assay.Results ① Under different concentrations of glucose,after treated with 10^-6-10^-5 mol·L^-1 rosiglitazone the proliferation of pancreatic β cells（NIT-1 cell line） was found（P0.05）and the apoptotic rate of cells was increased in a dose-dependent manner（1×10^-5 mol·L^-1 rosiglitazone group1×10^-6 mol·L^-1 rosiglitazone group1×10^-7 mol·L^-1 rosiglitazone group）.② When under same dose of glucose,the insulin secretion level in 11.1 mmol·L^-1 group was much higher than those in other groups（P〈0.05）,but the insulin secretion level was reduced gradually following the decrease of glucose concentration（11.1 mmol·L^-1 group16.7 mmol·L^-1 group22.5 mmol·L^-1 group27.6 mmol·L^-1 group）.The insulin secretion level in 5.6 mmol·L^-1 group was the lowest.③ After intervention of 10^-5 mol·L^-1rosiglitazone,the expression levels of both FOXO1 and TSC2 mRNA were significantly lower than those in control group（5.6 mmol·L^-1 group11.1 mmol·L^-1 group16.7 mmol·L^-1 group22.5 mmol·L^-1 group 27.6 mmol·L^-1 group）.When the glucose concentration was over 16.7 mmol·L^-1,the expressions of FOXO1 and TSC2 mRNA were obviously higher than those in the

关 键 词：葡萄糖 胰岛β细胞 结节性硬化症-2基因 叉头转录因子-1基因 罗格列酮 

分 类 号：R343[医药卫生—基础医学]