An in vitro senescence model of gingival epithelial cell induced by hydrogen peroxide treatment
An in vitro senescence model of gingival epithelial cell induced by hydrogen peroxide treatment
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DOI：
10.1007/s10266-021-00630-3
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发表时间：
2021-06-18
期刊：
影响因子：
2.5
通讯作者：
Furuichi, Yasushi
中科院分区：
文献类型：
--
作者：
Giri, Sarita;Takada, Ayuko;Furuichi, Yasushi
牙龈组织随着年龄的增长而逐渐变化，牙龈组织的体外模型可能有助于了解与年龄相关的口腔疾病。本研究旨在建立过氧化氢（H2O2）治疗模型来诱导人类牙龈上皮细胞衰老。此外，非瑟酮（一种研究抗衰老特性的黄酮类成分）也被用来检查它是否可以逆转诱导的衰老。培养原代人牙龈上皮祖细胞 (HGEPp) 并用不同浓度的 H2O2 处理。进行了细胞活力和形态、衰老相关 β-半乳糖苷酶 (SA-β-gal) 染色、已知衰老标志物 p16、p21 和 p53 的 mRNA 和蛋白质表达分析以及细胞周期测定。 H2O2 处理后，细胞的活力和形态、SA-β-gal 染色、相对 mRNA 和蛋白质表达以及细胞周期测定均显示出剂量依赖性变化。基于这些结果，400 μM H2O2 被认为是诱导衰老的最佳浓度。用非瑟酮处理衰老诱导的细胞下调了本研究中使用的所有衰老标记物。总之，建立了过氧化氢治疗诱导的牙龈上皮细胞衰老模型，可用于研究与年龄相关的牙周疾病。
Gingival tissue shows progressive changes with aging and an in vitro model of gingival tissue could be useful in understanding age-associated oral diseases. The present study aims to establish a hydrogen peroxide (H2O2) treatment model to induce aging in human gingival epithelial cells. In addition, fisetin, a flavonoid component studied for the anti-aging property is used to examine if it could reverse the induced senescence. Primary human gingival epithelial progenitor (HGEPp) cells were cultured and treated with different concentrations of H2O2. A cell vitality and morphology, senescence-associated beta-galactosidase (SA-beta-gal) staining, mRNA and protein expression analysis of known senescence markers p16, p21, and p53, and cell cycle assay were performed. The cells showed dose-dependent changes in vitality and morphology, SA-beta-gal staining, relative mRNA and protein expression, and cell cycle assay after H2O2 treatment. Based on these results, 400 mu M H2O2 was considered as an optimal concentration to induce senescence. Treatment of senescence-induced cells with fisetin downregulated all the senescence markers used in this study. In conclusion, a senescence model of gingival epithelial cells induced by hydrogen peroxide treatment was established which could be employed to study age-related periodontal diseases.