S-雌马酚改善高糖培养条件下INS-1细胞胰岛素分泌功能的研究  

S-equol ameliorates insulin secretion of INS-1 cells cultured in high glucose

作　　者：王丽[1] 陈卡[1] 刘凯[1] 高燕翔[1] 糜漫天[1] 

机构地区：[1]第三军医大学军事预防医学院营养与食品卫生学教研室、重庆市营养与食品安全重点实验室、重庆市医学营养研究中心,重庆400038

出　　处：《第三军医大学学报》2015年第5期386-391,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金(81102128)~~

摘　　要：目的研究大豆甙元活性代谢产物S-雌马酚(S-equol,S-Eq)对高糖培养条件下大鼠胰岛素瘤(INS-1)细胞胰岛素分泌功能的影响。方法采用26.2 mmol/L葡萄糖(高糖组,H)及葡萄糖分别与不同浓度S-Eq(0.1、1、10、100μmol/L S-Eq+H组)干预INS-1细胞24 h,另设未干预的INS-1细胞为对照组(C)。采用CCK-8检测细胞活力,ELISA法测定葡萄糖刺激胰岛素分泌(GSIS)功能,TUNEL法联合Annexin V-FITC/PI流式细胞术检测细胞凋亡,Real-time PCR和Western blot分别检测前胰岛素原(preproinsulin,PPI)、葡萄糖转运蛋白2(glucose transporter 2,Glut2)、线粒体阴离子载体解偶联蛋白2(uncoupling protein 2,UCP2)mRNA及蛋白表达。结果 S-Eq能显著增加高糖培养条件下INS-1细胞活力(P<0.05),其中1μmol/L S-Eq效果最为明显。GSIS检测发现,与对照组比较,高糖处理后INS-1细胞胰岛素分泌显著降低,而S-Eq能显著增加高糖处理的INS-1细胞的胰岛素分泌(P<0.05)。同时,0.1、1、10μmol/L S-Eq均能明显减少高糖培养条件下INS-1细胞凋亡,上调PPI mRNA、Glut2 mRNA和Glut2蛋白表达,而显著抑制UCP2 mRNA及其蛋白的表达(P<0.05)。结论 S-Eq可有效改善高糖培养条件下INS-1细胞胰岛素分泌功能,可能与抑制细胞凋亡,调节Glut2和UCP2表达有关。Objective To investigate the effect of S-equol（ S-Eq）,the active metabolite of daidzein,on the insulin secretion disorder induced by high glucose in rat insulinoma cell line INS-1. Methods INS-1cells were treated with 26. 2 mmol / L glucose in the absence or presence of different concentrations（ 0. 1,1,10 and 100 μmol / L） of S-Eq for 24 h. In this way,the cells were divided into 6 groups： control group（ C）,high glucose group（ H）,0. 1 μmol / L S-Eq ＋ high glucose group（ 0. 1 μmol / L S-Eq ＋ H）,1 μmol / L S-Eq＋ high glucose group（ 1 μmol / L S-Eq ＋ H）,10 μmol / L S-Eq ＋ high glucose group（ 10 μmol / L S-Eq ＋ H）,and 100 μmol / L S-Eq ＋ high glucose group（ 100 μmol / L S-Eq ＋ H）. The cell viability was examined by CCK-8 assay,and the glucose-stimulated insulin secretion（ GSIS） was tested with ELISA. INS-1 cell apoptosis was detected by TUNEL assay and Annexin V-FITC / PI flow cytometry. The mRNA expression of preproinsulin（ PPI）,glucose transporter 2（ Glut2）,and uncoupling protein 2（ UCP2） was tested by realtime PCR while the expression of Glut2 and UCP2 proteins was tested by Western blotting. Results S-Eq significantly increased the cell vitality of INS-1 cells stimulated by high glucose,and 1 μmol / L S-Eq had the most obvious effect（ P 〈0. 05）. The insulin secretion of INS-1 cells was significantly reduced by high glucose treatment,but S-Eq could remarkably increase insulin secretion in high glucose-treated INS-1 cells（ P 〈0. 05）. Meanwhile,0. 1,1 and 10 μmol / L of S-Eq reduced the apoptosis of high glucose-treated INS-1 cells,increased the expressions of PPI mRNA,Glut2 mRNA and protein,and significantly inhibited the expression of UCP2 at mRNA and protein levels（ P 〈0. 05）. Conclusion S-Eq can alleviate high glucose-induced insulin secretion disorder in INS-1 cells through inhibiting apoptosis and regulating the expressions of Glut2 and UCP2.

关 键 词：S-雌马酚 INS-1细胞 葡萄糖刺激胰岛素分泌 葡萄糖转运蛋白2 解偶联蛋白2 

分 类 号：R332[医药卫生—人体生理学] R151.2[医药卫生—基础医学]