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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100μmol/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m RNA水平;免疫印迹法检测RIG-I和细胞周期蛋白P27蛋白水平;Ed U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。
Objective: To investigate the role of retinoic acid inducible gene-1 (RIG-I) in the growth of mouse pancreatic β-cell line NIT-1 and its related molecular mechanisms. Methods: NIT-1 cells were stimulated with RIG-I specific agonist retinoic acid (RA) at different concentrations (1, 5, 10, 20, 50 and 100 μmol / L) RIG-I. MTT assay was used to detect cell viability; RIG-I m RNA level was detected by real-time PCR; RIG-I and P27 protein levels were detected by Western blotting; Ed U and flow cytometry were used to detect the proliferation and division cycle of β cells . RESULTS: Retinoic acid treatment increased the protein level of RIG-I in NIT-1 cells (P <0.05), and inhibited the growth of NIT-1 cells in a dose-dependent manner (P <0.05) And increased the level of P27 protein (P <0.05). CONCLUSION: RIG-I function can up-regulate the level of P27 protein in pancreatic islet β cells. Under certain conditions, it inhibits the proliferation of β-cell and induces cell cycle arrest in G1 phase, resulting in the loss of overall function of pancreatic β-cell. Type 2 diabetes is an important incentive for the development.