Long-term exposure to cigarette smoke influences characteristics in human gingival fibroblasts
Long-term exposure to cigarette smoke influences characteristics in human gingival fibroblasts
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DOI：
10.1111/jre.12891
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发表时间：
2021-05-31
影响因子：
3.5
通讯作者：
Murakami, Shinya
中科院分区：
文献类型：
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作者：
Tatsumi, Mari;Yanagita, Manabu;Murakami, Shinya
工作背景：牙周病是由牙周致病菌在龈沟和牙周袋内积聚引起的慢性炎症性疾病。吸烟是牙周病的一个公认的危险因素，吸烟者的牙周组织长期慢性暴露于香烟烟雾中。目的：在这项研究中，我们研究了长期暴露于尼古丁或香烟烟雾冷凝物（CSC）对人牙龈成纤维细胞（HGFs）细胞功能的影响。方法：将体外培养的HGF分为两组。短期和长期培养组的HGF分别在存在或不存在尼古丁的情况下培养4天和25天，尼古丁是香烟烟雾或CSC的主要成分之一。通过WST-1和伤口愈合试验评估暴露于尼古丁或CSC的HGF的细胞增殖和迁移能力。通过实时聚合酶链反应和酶联免疫吸附试验检查了暴露于尼古丁或CSC对各种细胞外基质（ECM）成分、炎症细胞因子和衰老相关基因表达的影响。通过衰老相关β-半乳糖苷酶（SA-β-gal）测定检测暴露于尼古丁或CSC的HGF的细胞衰老。为了探索衰老相关的microRNA（miRNA），我们从HGFs中提取miRNA，并通过miRNA阵列检测其表达谱。结果：在短期培养中，未观察到明显变化。HGFs长期暴露于尼古丁或CSC显著抑制其细胞增殖和迁移，并上调其类型。胶原蛋白类型。胶原、白细胞介素（IL）-6、IL-8、p16、p21和p53 mRNA表达以及IL-6和IL-8蛋白表达。此外，长期尼古丁或CSC暴露显著增加了SA-β-gal阳性HGF的百分比。此外，长期尼古丁或CSC暴露将miR-29 b和miR-199 a表达降低至未刺激HGF中的50%以下。结论：这些数据表明，长期吸烟习惯可能会降低伤口愈合能力，调节ECM蛋白稳态，刺激炎症反应，并加速HGF细胞衰老，从而加速牙周疾病的进展。
Background: Periodontal disease is a chronic inflammatory disease caused by peri-odontopathic bacteria accumulated in the gingival sulcus and periodontal pocket. Cigarette smoking is a well-established risk factor for periodontal disease, and periodontal tissues in smokers are chronically exposed to cigarette smoke on a long-term basis. Objective: In this study, we investigated the effects of long-term exposure to nicotine or cigarette smoke condensate (CSC) on cellular functions of human gingival fibroblasts (HGFs). Methods: In vitro-maintained HGFs were divided into two groups. The HGFs of the short-term and the long-term culture groups were cultured for 4 and 25 days, respectively, in the presence or absence of nicotine, which is one of the main components of cigarette smoke, or CSC. The cellular proliferation and migration capacities of HGFs exposed to nicotine or CSC were evaluated by WST-1 and wound healing assays. The effects of exposure to nicotine or CSC on the expression of various extracellular matrix (ECM) components, inflammatory cytokines, and senescence-related genes were examined by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The cellular senescence of HGFs exposed to nicotine or CSC was detected by the senescence-associated beta-galactosidase (SA-beta-gal) assay. To explore the senescence-associated microRNA (miRNA), we extracted miRNA from the HGFs and the expression profiles were examined by miRNA array. Results: In short-term culture, no significant changes were observed. Long-term exposure of HGFs to nicotine or CSC significantly suppressed their cellular proliferation and migration and upregulated type. collagen, type. collagen, interleukin (IL)-6, IL-8, p16, p21, and p53 mRNA expression, and IL-6 and IL-8 protein expression. Furthermore, long-term nicotine or CSC exposure significantly increased the percentage of SA-beta-gal-positive HGFs. In addition, long-term nicotine or CSC exposure reduced miR-29b and miR-199a expression to less than 50% of that in the unstimulated HGFs. Conclusion: These data suggest that long-term smoking habits may reduce wound healing ability, modulate ECM protein homeostasis, stimulate the inflammatory response, and accelerate cellular senescence in HGFs, and consequently accelerate the progression of periodontal diseases.