白藜芦醇预处理提高衰老牙周膜干细胞克隆和增殖潜能的研究

摘要

目的:观察白藜芦醇预处理能否衰使老牙周膜干细胞(Aged-PDLSCs)的增殖潜能和克隆形成能力得到增强.方法:将体外分离培养的中老年个体来源PDLSCs随机分为常规培养组、白藜芦醇预处理组、尼克酰胺预处理组;各组经相应处理后,采用MTT法检测其增殖潜能,结晶紫染色检测其克隆形成能力,RT-PCR和Western Blot检测其细胞中SIRT1和P53基因及蛋白的表达水平.结果:Aged-PDLSCs经白藜芦醇预处理后增殖潜能和克隆形成能力均得到了显著提高,而经尼克酰胺预处理后,其增殖潜能和克隆形成能力均明显降低;分别与常规培养组相比,差异均有统计学意义(P<0.05).此外,白藜芦醇预处理还可显著提高Aged-PDLSCs中SIRT1活性(P<0.05),显著降低了P53的表达水平(P<0.05).结论:白藜芦醇能够增强Aged-PDLSCs的克隆形成能力和增殖潜能,其作用可能是通过提高SIRT1的表达水平而实现的.%AIM: To investigate whether resveratrol(AV) can promote the proliferation and colony-forming of periodontal ligament stem cells (PDLSCs) derived from aged donors.METHODS: The PDLSCs were acquired from the periodontal ligament tissues of middle-aged and elderly subjects by enzymolysis and cultured in vitro.the PDLSCs were not pretreated(control group) or pretreated by AV (AV group) or nicotinamide(NT group).Cell proliferation was determined by MTT assay and colony formation was detected by crystal violet staining.PCR and Western blotting were performed to determine the relative expressions of SIRT1 and HIF1-α.RESULTS: AV pretreatment enhanced the capacities of proliferation and colony formation of PDLSCs derived from aged donors (P<0.05),while NT impaired the proliferation and colony formation of the PDLSCs (P<0.05).The expression of SIRT1 was promoted and the expression of P53 was down-regulated by AV pretreatment (P<0.05).CONCLUSION: Resveratrol can reverse the proliferation and colony formation of PDLSCs derived from aged donors via the activation of SIRT1.