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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。
Objective: To investigate the effect of ADAM28 gene on the proliferation, differentiation and apoptosis of human dental pulp stem cells (HDPSCs) and its possible mechanism. METHODS: Human dental pulp stem cells (HDPSCs) were isolated and cultured in vitro. The eukaryotic expression plasmid ADAM28 was constructed and transfected into HDPSCs. MTT colorimetric assay, enzyme kinetic assay and flow cytometry (FCM) ) To examine the effect of ADAM28 on the biological characteristics of HDPSCs. Immunocytochemistry and image analysis were used to detect the effect of ADAM28 on the expression of DSPP, BSP and OPN in HDPSCs. Statistical analysis was performed using SNK test using SPSS13.0 software package. Results: The proliferative activity and proliferative index of HDPSCs constructed and transfected with ADAM28 eukaryotic plasmid were significantly lower than those of empty vector group and untransfected group, and the secretion of alkaline phosphatase (ALP) and apoptotic cells (P <0.05). The expression of DSPP in HDPSCs was significantly increased (P <0.05). Conclusion: ADAM28 can significantly inhibit the proliferation of HDPSCs, promote the secretion activity of ALP and the expression of DSPP in HDPSCs, and induce the apoptosis of HDPSCs.