加热烟草制品（Ploom TECH）长期刺激对牙龈上皮细胞的影响。

Effects of prolonged stimulation with heated tobacco products (Ploom TECH ) on gingival epithelial cells.

机构信息

Division of Disease Control and Molecular Epidemiology, Department of Oral Growth and Development, School of Dentistry, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, 061-0293, Japan.

Advanced Research Promotion Center, Health Sciences University of Hokkaido, 1757 Kanazawa, Ishikari-Tobetsu, 061-0293, Japan.

出版信息

J Periodontal Res. 2023 Jun;58(3):553-563. doi: 10.1111/jre.13123. Epub 2023 Mar 27.

DOI:10.1111/jre.13123

PMID:36974375

Abstract

OBJECTIVE AND BACKGROUND

Heated tobacco products have recently become commercially available. These products, as well as combustible cigarettes, produce aerosols; the risk of various diseases associated with heated tobacco products may be the same or higher than that with combustible cigarettes. In this study, we examined the effect of Ploom TECH extract on gingival epithelial cells.

METHODS

Tobacco leaves from Ploom TECH tobacco capsules and water were mixed and heated; the supernatant subsequently collected was the heated tobacco product (HTP; control: HTP not added). Normal human gingival epithelial progenitors were cultured alternately with or without HTP for a total of 1 month. Subsequently, RNA, DNA, and proteins were isolated from these samples and comprehensively analyzed using RNA sequencing (RNA-seq), reduced representation bisulfite sequencing (RRBS), and western blotting, respectively.

RESULTS

RNA-seq revealed that 284 genes showed a twofold increase and 145 genes showed a twofold decrease in gene expression. A heat map showed genetic differences between the control and HTP groups. A principal component analysis plot showed a clear genetic distribution between the control and HTP. Gene Ontology (GO) analysis showed that genes related to seven GO terms, including cornification and keratinization, were induced by long-term HTP stimulation. By contrast, GO pathways with a significant decrease in component expression were not detected. RRBS revealed that CpG island methylation increased more than twofold in 158 genes and decreased to less than twofold in 171 genes. Methylation of these CpG islands was not correlated with changes in gene expression levels. HTP treatment increased S100A7 expression.

CONCLUSION

Long-term HTP stimulation affected epithelial differentiation and keratinization of gingival epithelial cells. Thus, habitual use of Ploom TECH may be a risk factor for tobacco-related oral mucosal diseases.

摘要

目的和背景

加热烟草制品最近已商业化。这些产品以及可燃香烟都会产生气溶胶；与加热烟草制品相关的各种疾病的风险可能与可燃香烟相同或更高。在这项研究中，我们研究了 Ploom TECH 提取物对牙龈上皮细胞的影响。

方法

将 Ploom TECH 烟弹中的烟草叶与水混合并加热；随后收集的上清液为加热烟草产品（HTP；对照：未添加 HTP）。将正常的人牙龈上皮祖细胞与 HTP 交替培养，共 1 个月。随后，从这些样品中分离出 RNA、DNA 和蛋白质，并分别使用 RNA 测序（RNA-seq）、简化代表性双测序（RRBS）和 Western 印迹进行综合分析。

结果

RNA-seq 显示 284 个基因的表达水平增加了两倍，145 个基因的表达水平降低了两倍。热图显示了对照组和 HTP 组之间的遗传差异。主成分分析图显示了对照组和 HTP 之间明显的遗传分布。基因本体论（GO）分析显示，与七个 GO 术语相关的基因，包括角蛋白化和角化，受到长期 HTP 刺激的诱导。相比之下，未检测到显著下调成分表达的 GO 途径。RRBS 显示，158 个基因的 CpG 岛甲基化增加了两倍以上，171 个基因的 CpG 岛甲基化减少了两倍以下。这些 CpG 岛的甲基化与基因表达水平的变化无关。HTP 处理增加了 S100A7 的表达。