口腔医学研究

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (5): 538-541.DOI: 10.13701/j.cnki.kqyxyj.2017.05.018

• 临床研究论著 • 上一篇    下一篇

IGF-1通过PI3K/AKT信号通路促进牙髓细胞增殖及碱性磷酸酶活性的研究

杜小颖1*,丁威薇2,陈悦3,迟丹丹1   

  1. 1. 大连市口腔医院综合急诊科 辽宁 大连 116021;2. 大连大学医学院口腔系 辽宁 大连 116622;
    3. 大连市妇幼保健院口腔科 辽宁 大连 116021
  • 收稿日期:2016-10-15 出版日期:2017-05-20 发布日期:2017-05-26
  • 通讯作者: 杜小颖,电话:13591118819
    [Key words] Dental pulp cell Cell proliferation
    Alkaline phosphatase


    牙髓细胞是牙髓组织中的主要细胞,具有一定的增殖和成骨分化能力,当牙髓组织受到外界刺激或损伤时,可刺激间充质细胞产生修复性牙本质,进而对牙髓组织进行保护和修复再生[1]。研究表明胰岛素样生长因子(IGF-1)参与调控牙髓细胞的增殖和分化过程,在机体生长发育过程中起重要作用[2,3]。IGF-1可通过JAK-STAT[4]等信号通路促进牙髓细胞增殖与分化,也有研究发现PI3K/AKT信号通路可促进人牙髓细胞的增殖和向牙本质分化[5],而关于IGF-1是否能够通过PI3K/AKT信号通路调控牙髓细胞增殖和分化未见报道。碱性磷酸酶(ALP)活性可反映牙髓细胞向牙本质细胞分化的能力。本研究旨在探讨PI3K/AKT信号通路在IGF-1促进牙髓细胞增殖和碱性磷酸酶活性中的作用,为牙髓组织的修复再生提供理论基础。
    1 材料与方法
  • 作者简介:杜小颖(1981~ ),女,辽宁大连人,硕士,主治医师,主要从事口腔内科的临床治疗工作。
  • 基金资助:
    大连市科技计划项目(编号:2013E15SF169)

Promotion of IGF-1 on the Proliferation and Alkaline Phosphatase Activity of Dental Pulp Cells through PI3K/AKT Signaling Pathway in Vitro.

DU Xiao-ying1, DING Wei-wei2, CHEN Yue3, CHI Dan-dan1.   

  1. 1. Department of Comprehensive Emergency, Dalian Stomatology Hospital, Dalian 116021, China; 2. Department of Oral Medicine, Medical College, Dalian University, Dalian 116622, China; 3. Department of Stomotology, Maternal and Child Health Hospital of Dalian, Dalian 116021, China.
  • Received:2016-10-15 Online:2017-05-20 Published:2017-05-26

PDF (PC)

319

被引次数

2

摘要： 目的:研究IGF-1对牙髓细胞增殖和碱性磷酸酶(ALP)活性及PI3K/AKT信号通路的影响。方法:采用酶消化法分离培养原代人牙髓细胞。Western blot检测牙髓组织中IGF-1蛋白表达,不同浓度的IGF-1处理牙髓细胞7d,CCK8法检测细胞增殖。用IGF-1(100 μg/L)及LY294002(10 μmol/L)分别单独或同时处理牙髓细胞,培养7 d后MTT实验检测细胞增殖;在培养第3、5、7、14天时检测细胞ALP 活性;在培养第7天时用Western blot检测AKT和p-AKT蛋白表达情况。结果:IGF-1在牙髓炎组织中低表达, IGF-1在20~100 μg/L浓度范围内从作用第3天开始能够显著促进牙髓细胞增殖(P<0.05或P<0.01),且具有剂量和时间依赖效应。LY294002能够抑制牙髓细胞的增殖和碱性磷酸酶活性,具有时间依赖性。IGF-1能促进p-AKT蛋白表达,而 LY294002能减低IGF-1对p-AKT蛋白表达的促进作用。结论:IGF-1可以促进牙髓细胞增殖和碱性磷酸酶活性,且具有浓度和时间依赖性,作用机制可能与PI3K/AKT信号通路相关。
[关键词] 牙髓细胞 细胞增殖 碱性磷酸酶

关键词: 牙髓细胞, 细胞增殖, 碱性磷酸酶

Abstract: Objective: To study the effect of IGF-1 on the proliferation and alkaline phosphatase activity of dental pulp cells and its influences on PI3K/AKT signal pathway. Methods: Primary human dental pulp cells were isolated and cultured by enzymatic digestion. IGF-1 protein expression in dental pulp tissue was detected by Western blot. The dental pulp cells were then treated with IGF-1 at different concentrations, and cell proliferation was detected by MTT assay after 7 days. The dental pulp cells were treated with IGF-1 (100ng/mL) and LY294002 (10μmol/L) respectively or simultaneously for 7 days, and cell proliferation was detected by MTT assay after 7 days. ALP activity was detected after culture for 3, 5, 7 and 14 days. The protein expression of AKT and p-AKT was detected after culture for 7days. Results: IGF-1 showed Low expression in pulpitis. IGF-1 could significantly promote the proliferation of dental pulp cells in the concentration range of 20-100 ng/mL since 3rd day(P<0.05)with a dose- and time-dependent effect. LY294002 could inhibit the proliferation of dental pulp cells and alkaline phosphatase activity with time-dependence. IGF-1 could promote the expression of p-AKT protein, while LY294002 could reduce the effect of IGF-1 on the expression of p-AKT protein. Conclusion: IGF-1 can promote the proliferation and alkaline phosphatase activity of dental pulp cells in concentration and time dependence manner, which may be related to the PI3K/AKT signaling pathway.

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