精氨酸代琥珀酸合成酶对胰岛β细胞增殖与凋亡的影响
DOI:
作者:
作者单位:

1.南京医科大学第一附属医院;2.盐城市第三人民医院

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金项目(面上项目,重点项目,重大项目)


Effects of argininosuccinate synthetase 1 on cell proliferation and apoptosis of pancreatic islet β cells
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    摘要: 目的:探讨精氨酸代琥珀酸合成酶(ASS1)对胰岛β细胞增殖与凋亡的影响及机制。方法:利用siRNA和慢病毒载体在胰岛β-TC6细胞中分别敲低和过表达ASS1;5-乙炔基-2'-脱氧尿嘧啶核苷(EdU)和Cell counting kit-8(CCK-8)检测细胞增殖能力;Annexin V-PI流式细胞术和TUNEL检测细胞凋亡水平;Western blot检测B细胞淋巴瘤-2(Bcl-2),Bcl-2相关X蛋白(Bax),半胱氨酸天冬氨酸酶3(Caspase3),切割型半胱氨酸天冬氨酸蛋白水解酶-3(Cleaved Caspase-3),凋亡诱导因子(AIF)、细胞核增殖抗原(Ki67)和哺乳动物雷帕霉素靶蛋白(mTOR)表达水平;RT-qPCR检测凋亡诱导因子(AIF)、细胞核增殖抗原(Ki67)和哺乳动物雷帕霉素靶蛋白(mTOR)mRNA水平。组间数据比较采用单因素方差分析。结果:1.与对照相比,敲低ASS1后,胰岛β细胞EdU阳性率和CCK-8细胞增殖活力降低,TUNEL阳性细胞数和AV/PI检测的细胞凋亡率升高。胰岛β细胞内AIF表达量明显升高,Bax/Bcl-2比值下降,Caspase-3活性降低。2.过表达ASS1后,TUNEL阳性细胞数和AV/PI检测的细胞凋亡率均较对照组降低,伴随胰岛β细胞内Ki67和mTOR表达量升高。但胰岛β细胞EdU阳性率和CCK-8细胞增殖活力无明显变化。结论:ASS1过表达可能激活mTOR信号通路促进胰岛β细胞增殖;ASS1表达降低时,胰岛β细胞可通过AIF途径启动细胞的凋亡并负反馈抑制了Bax和Caspase-3的活性。ASS1在胰岛β细胞的增殖和凋亡中发挥一定调控作用。

    Abstract:

    Abstract: Objective: To investigate the effect of argininosuccinate synthetase 1 (ASS1) on cell proliferation and apoptosis of pancreatic islet β cells. Methods: Small interference RNA was used to knockdown ASS1, and lentiviral vector to overexpress ASS1 in pancreatic islet β cells. 5‐ethynyl‐2′‐deoxyuridine (EdU) and cell counting kit-8 (CCK-8) to detect proliferation ability, Annexin V-PI flow cytometry and terminal dUTP nick-end labeling (TUNEL) to detect apoptosis. Western blot was used to examined protein expression of Bcl-2 (B-cell leukaemia-2), Bax (Bcl-2 Associated X Protein), Caspase-3, Cleaved Caspase-3, apoptosis inducing factor (AIF), nuclear proliferation antigen Ki67 and mammalian rapamycin target protein (mTOR). RT-qPCR was applied to detect mRNA expression of AIF, Ki67 and mTOR. Data between groups was compared with one-way ANOVA. Results: 1. Proliferation activity of pancreatic β cells detected by EdU and CCK-8 was lower when ASS1 was knockdown. and apoptosis of islet β cells examined by TUNEL method and flow cytometry was higher than that in NC group; 2. Apoptosis of β cells decreased when ASS1 was overexpressed. Meanwhile, increased AIF expression and decreased ratio of Bax/Bcl-2 and the activity of Caspase-3 were observed. Conclusion: Overexpressing ASS1 in pancreatic islet β cells may activate mTOR signaling pathway to promote cell proliferation. Pancreatic islet β cells with decreased ASS1 expression may initiate apoptosis through AIF pathway and inhibit the activities of Bax and Caspase-3 by negative feedback. In summary, ASS1 plays a role in pancreatic islet β-cell’s proliferation and apoptosis.

    参考文献
    相似文献
    引证文献
引用本文
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2022-11-09
  • 最后修改日期:2023-01-13
  • 录用日期:2023-05-23
  • 在线发布日期:
  • 出版日期: