人牙髓干细胞增殖与瞬时受体电位M7通道的作用  

Role of transient receptor potential melastatin 7 in the proliferation of human dental pulp stem cells

作　　者：崔力[1,2] 徐帅妹[1,2] 吴补领[1,2] 

机构地区：[1]南方医科大学南方医院,广东省广州市510515 [2]南方医科大学口腔医学院,广东省广州市510515

出　　处：《中国组织工程研究》2013年第19期3495-3502,共8页Chinese Journal of Tissue Engineering Research

基　　金：广东省高等学校人才引进科研资助课题项目(C1010047);项目名称为牙胚发生位置;大小;形态的蛋白质组学研究和牙再生~~

摘　　要：背景:瞬时受体电位M7通道广泛存在于机体组织和细胞,对细胞存活、增殖和维持细胞镁离子平衡是必须的。目的:探讨瞬时受体电位M7通道对人牙髓干细胞增殖的作用。方法:酶消化法分离培养人牙髓干细胞,运用不同浓度(50,100μmol/L)瞬时受体电位M7通道抑制剂2-氨基乙基二苯硼酸酯作用于人牙髓干细胞72h后,Western-blot检测2-氨基乙基二苯硼酸酯对瞬时受体电位M7通道蛋白水平的影响,MTT法观察其对人牙髓干细胞增殖的影响;构建特异抑制瞬时受体电位M7通道表达慢病毒载体,运用慢病毒感染人牙髓干细胞。进行RT-PCR和Western-blot沉默效果分析,在1,3,5,7d进行MTT法检测特异沉默瞬时受体电位M7通道后对细胞增殖的影响。结果与结论:50,100μmol/L2-氨基乙基二苯硼酸酯均能抑制瞬时受体电位M7通道蛋白水平的表达并且能明显抑制人牙髓干细胞增殖(P<0.01),特异性沉默瞬时受体电位M7通道表达后,在不同时间段人牙髓干细胞增殖能力均明显降低(P<0.01)。提示瞬时受体电位M7通道对维持人牙髓干细胞增殖能力有重要作用。BACKGROUND：Transient receptor potential melastatin 7 channel is widely distributed in tissues and cel s, and it is necessary for cel survival, proliferation and maintenance of magnesium ion balance. OBJECTIVE：To investigate the role of transient receptor potential melastatin 7 channel in the proliferation of human dental pulp stem cel s. METHODS：Human dental pulp stem cel s were isolated and cultured by col agenase digestion method, and were exposed for 72 hours to 2-aminoethoxydiphenyl borate （50 and 100μmol/L）, which was widely used as an inhibitor of transient receptor potential melastatin 7. The influences of 2-aminoethoxydiphenyl borate on transient receptor potential melastatin 7 protein level and proliferation of human dental pulp stem cel s were detected with western blot assay and 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide method, respectively. Specific lentivirus vector was constructed to inhibit the expression of transient receptor potential melastatin 7, and lentivirus was applied to transfect human dental pulp stem cel s. Reverse transcription-PCR and western blot for silencing efficacy analysis were also performed. Changes of cel proliferation after specific silencing transient receptor potential melastatin 7 was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay at 1, 3, 5, and 7 days after culture. RESULTS AND CONCLUSION：The proliferation of human dental pulp stem cel s and protein expression of transient receptor potential melastatin 7 were significantly inhibited by 2-aminoethoxydiphenyl borate at 50 and 100μmol/L （P〈0.01）. After specific silencing transient receptor potential melastatin 7, the proliferation capacity of human dental pulp stem cel s was significantly decreased at different time poinrs （P〈0.01）. Transient receptor potential melastatin 7 plays an important role in maintaining proliferation ability of human dental pulp stem cel s.

关 键 词：干细胞 干细胞培养与分化 瞬时受体电位M7通道 人牙髓干细胞 SHRNA 慢病毒 2-氨基乙基二苯硼酸酯 增殖 省级基金 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]