摘要
目的:探讨金属蛋白酶解离素28(ADAM28)基因对人牙髓干细胞(HDPSCs)增殖、分化、凋亡特性的影响和可能的作用机制。方法:体外分离培养、鉴定人牙髓干细胞(HDPSCs),应用基因重组构建ADAM28真核表达质粒并转染HDPSCs,应用四唑盐(MTT)比色法、酶动力学法和流式细胞术(FCM)检测ADAM28对HDPSCs生物学特性的影响。应用免疫细胞化学和图像分析技术检测ADAM28对HDPSCs表达牙本质涎磷蛋白(DSPP)、骨涎蛋白(BSP)、骨桥蛋白(OPN)的影响。采用SPSS13.0软件包的SNK检验进行统计学分析。结果:成功构建并转染ADAM28真核质粒,真核质粒转染组HDPSCs的增殖活性、增殖指数显著低于空载体组和未转染组,碱性磷酸酶(ALP)分泌水平及凋亡细胞百分比明显上升,差异显著(P<0.05);HDPSCs内DSPP的表达水平显著提高(P<0.05)。结论:ADAM28可显著抑制HDPSCs增殖,促进ALP的分泌活性和HDPSCs内DSPP的表达,显著诱导HDPSCs的凋亡。
PURPOSE： To investigate the effects of a disintegrin and metalloproteinase 28 （ADAM28） on proliferation, differentiation and apoptosis of human dental pulp stem cells （HDPSCs） and the possible mechanism. METHODS： Firstly, HDPSCs were isolated and cultured in vitro and identified. ADAM28 eukaryotic expression plasmid was constructed via gene rebuilt technique and transfected into HDPSCs. Then MTT chromatometry, enzyme dynamics and flow cytometry （FCM） techniques were performed to detect the effects of ADAM28 on biological characteristics of HDPSCs. Immunocytochemical and image analysis techniques were used to determine the influence of ADAM28 on HDPSCs expressing dentin sialophosphoprotein （DSPP）, bone sialoprotein （BSP） and osteopontin （OPN）. Statistical significance was assessed by the Student-Newman-Keuls （SNK） test with SPSS 13.0 software package. RESULTS： ADAM28 eukaryotic plasmid was constructed and transfected into HDPSCs for 48 hours successfully. In ADAM28 eukaryotic plasmid group, proliferation activity and index of HDPSCs were lower than those of pcDNA3.1 （＋） group and untransfected group significantly.Alkaline phosphatase （ALP） secretion level and percentage of apoptotic cells went up remarkly. Significant difference was detected between eukaryotic plasmid group and other groups （P0.05）. The expression level of DSPP in HDPSCs elevated significantly （P0.05）. CONCLUSIONS： ADAM28 could inhibit HDPSCs proliferation, promote ALP secretion activity and DSPP expression in HDPSCs and induce HDPSCs apoptosis significantly. Supported by National Natural Science Foundation of China （Grant No.30772450）.
出处
《上海口腔医学》
CAS
CSCD
2010年第3期262-269,共8页
Shanghai Journal of Stomatology
基金
国家自然科学基金(30772450)
