DNA甲基化酶抑制剂5-Aza-CdR对人牙髓细胞增殖活性及矿化能力的影响  

Effect of DNA methyltransferase inhibitor 5-aza-2'-deoxytidine on the proliferation and differentiation of human dental pulp cells in vitro

作　　者：张德茜[1] 李启梦[1] 徐琼[1] 

机构地区：[1]中山大学光华口腔医学院.附属口腔医院.口腔医学研究所广东省口腔医学重点实验室,广东广州510055

出　　处：《牙体牙髓牙周病学杂志》2013年第10期618-621,626,共5页Chinese Journal of Conservative Dentistry

基　　金：广东省大学生创新训练项目(1055812388)

摘　　要：目的：研究DNA甲基化酶抑制剂5-氮杂脱氧胞苷（5-Aza-2＇-deoxycytidine,5-Aza-CdR）对人牙髓细胞（human dental pulp cells,hDPCs）增殖活性和矿化能力的影响.方法：以酶消化联合组织块法体外培养人牙髓细胞,取第3代细胞并随机分为常规培养组（对照组）、矿化诱导组、5-Aza-CdR组及5-Aza-CdR联合矿化诱导液组（联合组）；分别于培养不同时间后,采用CCK8法检测各组hDPCs细胞的增殖活性；碱性磷酸酶活性检测法以及茜素红矿化结节染色法观察各组hDPCs的矿化能力.结果：与对照组相比,矿化诱导组、5-Aza-CdR组及联合组在培养第3～7天时,hDPCs的增殖活性明显降低（P＜0.05）；第3～14天时,碱性磷酸酶活性明显升高（P＜0.05）.第14天时,除对照组外各组均有矿化结节形成,其中5-Aza-CdR组矿化结节较少,矿化诱导组及联合组可见大片矿化结节,特别是联合组,矿化结节的密度更大、颜色更深.结论：体外培养条件下,DNA甲基化酶抑制剂5-Aza-CdR可降低hDPCs的增殖活性,增强其矿化能力.AIM：To investigate the effects of DNA methyltransferases inhibitors 5-aza-2＇-deoxytidine （5-Aza-CdR） on the proliferation and differentiation of human dental pulp cells （hDPCs） in vitro.METHODS：hDPCs were isolated and cultured from extracted healthy third molars and premolars.The cells of third passage were divided as common culture group （group 1）,osteogenic induction group （group 2）,5-Aza-CdR treatment group （group 3） and osteogenic induction combined with 5-Aza-CdR treatment group（group 4）.At different time points after culture,CCK8 method was used to quantify the cell proliferation activity.Alkaline phosphatase （ALP）quantity and Alizarin red staining were used to observe the differentiation of hDPCs.RESULTS：The hDPCs in group 2,3 and 4 showed lower proliferation activity and higher ALP activity than in group 1 from day 3 to day 14 of cultyre （P ＜0.05）.Alizarin Red staining on day 14 showed greater calcification in group 2 and 4 than in group 3,while negative staining was found in group 1.CONCLUSION：5-Aza-CdR can inhibit the proliferation and stimulate the differentiation of hDPCs in vitro.

关 键 词：人牙髓细胞 5-Aza-2＇-deoxycytidine 分化 增殖 

分 类 号：R780.2[医药卫生—口腔医学]