CAJ | 학술논문

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72 h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1,3,5,7 d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/L 2-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.
배경:순시수체전위M7통도엄범존재우궤체조직화세포,대세포존활、증식화유지세포미리자평형시필수적.목적:탐토순시수체전위M7통도대인아수간세포증식적작용.방법:매소화법분리배양인아수간세포,운용불동농도(50,100μmol/L)순시수체전위M7통도억제제2-안기을기이분붕산지작용우인아수간세포72 h후,Western-blot 검측2-안기을기이분붕산지대순시수체전위M7통도단백수평적영향,MTT법관찰기대인아수간세포증식적영향;구건특이억제순시수체전위M7통도표체만병독재체,운용만병독감염인아수간세포.진행RT-PCR화Western-blot침묵효과분석,재1,3,5,7 d진행MTT법검측특이침묵순시수체전위M7통도후대세포증식적영향.결과여결론:50,100μmol/L 2-안기을기이분붕산지균능억제순시수체전위M7통도단백수평적표체병차능명현억제인아수간세포증식(P <0.01),특이성침묵순시수체전위M7통도표체후,재불동시간단인아수간세포증식능력균명현강저(P <0.01).제시순시수체전위M7통도대유지인아수간세포증식능력유중요작용.
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.