人牙龈间充质干细胞条件培养液对人成纤维细胞生物学功能的影响

Effect of human gingival mesenchymal stem cells derived conditioned medium on the biological function of dermal fibroblasts

目的 观察人牙龈间充质干细胞条件培养液(GMSCs-CM)对人成纤维细胞生物学功能的影响。方法 制备GMSCs-CM,将0(空白对照组)、1、5、10、50、100μg/mL浓度的GMSCs-CM作用于成纤维细胞,采用CCK-8法检测成纤维细胞增殖情况,TUNEL法检测成纤维细胞凋亡情况,实时荧光定量PCR(qRT-PCR)检测在0(空白对照组)、1、10、100μg/mL浓度的GMSCs-CM作用下成纤维细胞的Ⅰ型胶原(COL-1)、增殖细胞核抗原(PCNA)、α-平滑肌动蛋白(α-SMA)、转化生长因子β(TGF-β)mRNA的表达。结果 GMSCs-CM能明显促进成纤维细胞的增殖。当GMSCs-CM浓度为1μg/mL时促进成纤维细胞增殖的作用最为显著,而对细胞凋亡无明显影响。与空白对照组比较,GMSCs-CM浓度为1μg/mL明显提高成纤维细胞COL-1、PCNA、TGF-β mRNA表达水平(均P<0.05)。结论 GMSCs-CM对成纤维细胞的增殖能力有明显的促进作用,而对细胞凋亡无明显影响,有望用于促进皮肤创面愈合治疗。

Objective To investigate the effect of human gingival mesenchymal stem cells derived conditioned medium(GMSCs-CM)on the biological function of human fibroblasts.Methods Human dermal fibroblasts were cultured with 0,1,5,10,50 and 100μg/mL of GMSCs-CM,respectively.The proliferation of fibroblasts was assessed with CCK-8 assay;the apoptosis of fibroblasts was measured with TUNEL assay;the mRNA expressions of type I collagen(COL-1),proliferating cell nuclear antigen(PCNA),α-smooth muscle actin(α-SMA)and transforming growth factor-β(TGF-β)in fibroblasts were detected in GMSCs-CM 0,1,5,10,100μg/mL with quantitative real-time PCR.Results GMSCs-CM significantly promoted the proliferation of dermal fibroblasts,and the concentration of 1μg/mL exhibited the most significant effect on fibroblast proliferation.GMSCs-CM did not affect the apoptosis of fibroblasts.GMSCs-CM increased the expression levels of COL-1,PCNA,and TGF-βmRNA in dermal fibroblasts(all P<0.05).Conclusion GMSCs-CM can enhance the proliferation of dermal fibroblasts without effect on the cell apoptosis,indicating that GMSCs-CM may promoting skin wound healing.