RIG-I上调抑制胰腺β细胞增殖机制研究
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国家自然科学基金(面上项目81170714)


Effects of retinoic acid inducible gene-I on the proliferation of pancreatic β-cell
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    摘要:

    目的:探讨维甲酸诱导基因I(retinoic acid inducible gene-I,RIG-I)在小鼠胰岛β细胞株NIT-1生长中的作用及其相关分子机制-方法:应用不同浓度(1-5-10-20-50和100 μmol/L)的RIG-I特异性激动剂维甲酸(retinoic acid,RA)分别刺激NIT-1细胞6-12-24 h,上调RIG-I-MTT法检测细胞生长活力;real-time PCR测定RIG-I的 mRNA水平;免疫印迹法检测RIG-I和细胞周期蛋白P27蛋白水平;EdU和流式细胞术检测β细胞的增殖和分裂周期-结果:维甲酸处理能刺激NIT-1细胞RIG-I蛋白水平增加(P < 0.05),能剂量依赖地抑制NIT-1细胞生长活力(P < 0.05),且诱导NIT-1细胞G1期阻滞和周期抑制蛋白P27蛋白水平增加(P < 0.05)-结论:RIG-I的功能增强能诱导小鼠胰岛β细胞P27蛋白水平上调,一定条件下抑制β细胞增殖和诱导细胞周期G1期阻滞,从而导致机体胰岛β细胞整体功能的失代偿,可能是2 型糖尿病发生发展的重要诱因-

    Abstract:

    Objective:To investigate the effect of elevated retinoic acid inducible gene-I (RIG-I)on the growth of NIT-1 pancreatic β-cell and possible mechanisms during this process. Methods:Different concentrations of retinoic acid (RA)were used to activate RIG-I with different periods. Cell viability was assessed by MTT colorimetric assay. RIG-I was detected using real-time RT-PCR and Western blot analysis. Cell cycle distribution was measured by FACS analysis,and cell cycle protein p27 was further detected by Western blot analysis. Results:Retinoic acid upregulated the mRNA and protein level of RIG-I in NIT-1 cells (P < 0.05). Elevated RIG-I significantly inhibited the viability of NIT-1 cells in a dose-dependent manner (P < 0.05). Furthermore,elevated RIG-I induced an increase in the proportion of cells at the G1 phase and a corresponding decrease in the number of cells at the S phase (P < 0.05). Additionally,elevated RIG-I caused a significant increase of p27 protein (P < 0.05). Conclusion:The results suggest that activated RIG-I induced upregulation of p27 in pancreatic β-cell,which inhibits proliferation through cell cycle arrest at the G1 phase,and ultimately results in an overall β-cell mass deficiency.

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潘 漪,高丽丽,王鼎玉,陈婷婷,郭 军. RIG-I上调抑制胰腺β细胞增殖机制研究[J].南京医科大学学报(自然科学版),2016,(5):564-568

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  • 收稿日期:2016-01-07
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  • 在线发布日期: 2016-05-23
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