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Fox O1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/m L青霉素、100 U/m L链霉素)于25 m L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100μIU/m L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Fox O1的表达位置变化,用Image pro plus软件对Fox O1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Fox O1荧光强度逐渐增强,而细胞核Fox O1荧光强度减弱(P<0.05)。结论:高浓度胰岛素孵育min6细胞使Fox O1出细胞核转位至细胞质,并且具有时间依赖性,提示Fox O1是高胰岛素血症对β细胞功能影响的机制之一。
Objective: To explore the effect of exogenous high insulin on Fox O1 nucleus--cytoplasmic shutting in β cells. Methods: 1. The min6 cells were cultured with DMEM( low glucose) medium( containing 15 % FBS, 100 U/m L penicillin, 100U/m L streptomycin) in a 25 m L culture flask under the condition of 37℃, 5 % CO2. 2. Making min6 cells growing on glass coverslips, then stimulating the cells with 100 μIU/m L insulin in 12 h, 24 h and 48 h separately. 3. The expression of Fox O1 in min6 cells in different incubation time exposure to 100 μIU/m L insulin was analyzed by immune fluorescence cytochemistry combined with laser scanning confocal microscopy. Semi-quantitative analysis of fluorescence intensity of Fox O1 was determined by Image Pro Plus 6.0 software. Results: Compared with the control group, the fluorescence intensity of Fox O1 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). Conclusions: High concentration of insulin could induce Fox O1 translocation from nucleus to the cytoplasm in a time-dependent manner, suggesting that Fox O1 may be one of the mechanisms of the impact of hyperinsulinemia on β-cell function.
出处
《现代生物医学进展》
CAS
2015年第29期5621-5623,5644,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(30971122)