文章摘要
雷雨欣 焦凯 赵国宏 卫静 杨璐 王婷.胰岛素对beta细胞FoxO1 胞质-胞核穿梭定位的影响[J].,2015,15(29):5621-5623
胰岛素对beta细胞FoxO1 胞质-胞核穿梭定位的影响
Effect of Insulin on FoxO1 Nucleus-cytoplasmic Shuttling in beta Cells
  
DOI:
中文关键词: FoxO1  胰岛素  min6 细胞
英文关键词: FoxO1  Insulin  Min6 cells
基金项目:国家自然科学基金项目(30971122)
作者单位
雷雨欣 焦凯 赵国宏 卫静 杨璐 王婷 第四军医大学唐都医院内分泌科 
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中文摘要:
      目的:通过激光扫描共聚焦显微镜对小鼠胰岛素瘤min6 细胞免疫化学染色后观察外源性高胰岛素对FoxO1 胞质- 胞核穿 梭定位的影响。方法:小鼠胰岛素瘤min6 细胞用DMEM( low glucose )培养基(含有15%FBS 、100 U/mL青霉素、100 U/mL链霉 素)于25 mL培养瓶放置在37 ℃、5%CO2浓度的细胞孵箱中培养。细胞爬片后给予100 uIU/mL 浓度胰岛素分别刺激12、24 和 48 小时。细胞免疫化学染色后激光扫描共聚焦显微镜观察FoxO1 的表达位置变化,用Image pro plus 软件对FoxO1 荧光强度进 行半定量分析。结果:与低糖孵育的对照组相比,胰岛素孵育12 h、24 h和48 h时胞质内FoxO1 荧光强度逐渐增强,而细胞核 FoxO1 荧光强度减弱(P<0.05)。结论:高浓度胰岛素孵育min6细胞使FoxO1 出细胞核转位至细胞质,并且具有时间依赖性,提 示FoxO1是高胰岛素血症对beta细胞功能影响的机制之一。
英文摘要:
      Objective:To explore the effect of exogenous high insulin on FoxO1 nucleus--cytoplasmic shutting in beta cells.Methods:1. The min6 cells were cultured with DMEM( low glucose ) medium ( containing 15 %FBS, 100 U/mL penicillin, 100U/mL streptomycin ) in a 25 mL culture flask under the condition of 37℃, 5 %CO2. 2. Making min6 cells growing on glass coverslips, then stimulating the cells with 100 uIU/mL insulin in 12 h, 24 h and 48 h separately. 3. The expression of FoxO1 in min6 cells in different incubation time exposure to 100 uIU/mL insulin was analyzed by immune fluorescence cytochemistry combined with laser scanning confocal microscopy. Semi-quantitative analysis of fluorescence intensity of FoxO1 was determined by Image ProPlus 6.0 software.Results:Compared with the control group, the fluorescence intensity of FoxO1 which incubated with insulin for 12 h, 24 h and 48 h of min6 cells was gradually increased in cytoplasm and decreased in nucleus (P<0.05).Conclusion:High concentration of insulin could induce FoxO1 translocation fromnucleus to the cytoplasmin a time-dependent manner, suggesting that FoxO1 may be one of the mechanisms of the impact of hyperinsulinemia on beta-cell function.
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