人牙龈成纤维细胞复制性衰老模型的构建

发布时间：2018-04-10 13:20

  本文选题：牙龈 + 细胞衰老　； 参考：《上海交通大学学报(医学版)》2017年05期

【摘要】：目的·构建人牙龈成纤维细胞(hGFs)复制性衰老模型,探讨衰老相关生物学改变,为牙周疾病衰老相关研究提供有效的细胞模型。方法·取健康牙龈组织,利用组织块法体外培养hGFs,免疫荧光染色验证组织来源;连续传代培养,计算累积细胞群体倍增指数(CPD)并绘制曲线;取CPD值为10.82、20.65、29.52、42.22、60.79、70.03的hGFs,CCK-8试剂盒检测细胞增殖能力改变,real-time PCR检测衰老相关基因p16~(INK4a)、p21~(Cip1)的表达变化。结果·连续传代培养后,CPD值逐渐增大,达70.03时进入平台期;随着CPD值的增大,hGFs细胞形态呈扁平状,细胞突起增多;细胞增殖能力减弱;p16~(INK4a)、p21~(Cip1) mRNA表达水平显著升高(P=0.000)。结论·连续传代培养可构建hGFs复制性衰老模型,CPD曲线可反映细胞衰老状态。
[Abstract]:Objective to establish a replicating aging model of human gingival fibroblasts (hGFs) and explore the biological changes associated with senescence in order to provide an effective cell model for the study of aging of periodontal diseases.Methods healthy gingival tissue was cultured in vitro with tissue mass method, and the tissue origin was verified by immunofluorescence staining, then the cumulative cell population doubling index (CPD) was calculated and the curves were drawn.The expression of the senescence related gene p16NK4a was detected by real-time PCR with a CPD value of 10.82n 20.655.52 (42.2260.709 / 70.03) and a CCK-8 kit of hGFsfen CCK-8 was used to detect the change of cell proliferative ability and the expression of the senescence related gene p16NK4a (p21 ~ (1)) was detected by real-time PCR.Results after continuous passage culture, the CPD value increased gradually, reaching 70.03 to the plateau stage, with the increase of CPD value, the morphology of hGFs cells was flat, the cell processes increased, and the cell proliferation ability decreased. The expression level of P0.000 mRNA was significantly increased.Conclusion continuous passage culture can construct hGFs replicative senescence model and the curve can reflect the state of cell senescence.
【作者单位】： 上海交通大学医学院附属第九人民医院牙周病科上海市口腔医学重点实验室;
【基金】：国家自然科学基金(81570977) 上海市教育委员会高峰高原学科建设计划~~
【分类号】：R781.4
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本文编号：1731365