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  

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

作　　者：杜小颖 丁威薇[2] 陈悦 迟丹丹 

机构地区：[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

出　　处：《口腔医学研究》2017年第5期538-541,共4页Journal of Oral Science Research

基　　金：大连市科技计划项目(编号:2013E15SF169)

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(100μg/L)及LY294002(10μmol/L)分别单独或同时处理牙髓细胞,培养7d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20~100μg/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。Objective：To study the effect of IGF-1on 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-1protein expression in dental pulp tissue was detected by Western blot.The dental pulp cells were then treated with IGF-1at different concentrations,and cell proliferation was detected by MTT assay after 7days.The dental pulp cells were treated with IGF-1（100ng/mL）and LY294002（10μmol/L）respectively or simultaneously for 7days,and cell proliferation was detected by MTT assay after 7days.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-1showed Low expression in pulpitis.IGF-1could significantly promote the proliferation of dental pulp cells in the concentration range of 20-100ng/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-1could 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-1can 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.

关 键 词：牙髓细胞 细胞增殖 碱性磷酸酶 

分 类 号：R781.3[医药卫生—口腔医学]