二十碳五烯酸对人牙龈成纤维细胞生物学活性及炎症因子表达的影响

doi:10.19439/j.sjos.2021.03.001

上海口腔医学 ›› 2021, Vol. 30 ›› Issue (3): 225-231.doi: 10.19439/j.sjos.2021.03.001

二十碳五烯酸对人牙龈成纤维细胞生物学活性及炎症因子表达的影响

周可聪, 孙梦君, 夏一如, 谢玉峰, 束蓉

上海交通大学医学院附属第九人民医院牙周病科,上海交通大学口腔医学院,国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海 200011

收稿日期:2020-11-09 修回日期:2021-01-06 出版日期:2021-06-25 发布日期:2021-08-05
通讯作者: 束蓉,E-mail：shurong1977@163.com
作者简介:周可聪（1996-）,女,硕士研究生,住院医师,E-mail：zhoukecong111@126.com
基金资助:
国家自然科学基金重大项目（81991500、81991503）; 口腔颌面组织再生与功能修复创新团队（SSMU-ZDCX20180900）; 上海交通大学医学院附属第九人民医院交叉基金（JYJC201904）; 上海交通大学医学院附属第九人民医院基础研究助推计划（JYZZ108）

Effects of eicosapentaenoic acid on biological activity and inflammatory factor expression of human gingival fibroblasts

ZHOU Ke-cong, SUN Meng-jun, XIA Yi-ru, XIE Yu-feng, SHU Rong

Department of Periodontology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology. Shanghai 200011, China

Received:2020-11-09 Revised:2021-01-06 Online:2021-06-25 Published:2021-08-05

摘要/Abstract

摘要： 目的: 探讨二十碳五烯酸（eicosapentaenoic acid,EPA）对人牙龈成纤维细胞（human gingival fibroblasts,HGFs）生物学活性及炎症因子表达的影响。方法: 分别通过活-死细胞染色、免疫荧光染色、流式细胞术观察EPA对HGFs细胞活性、形态、细胞周期的影响,采用牙龈卟啉单胞菌（Porphyromonas gingivalis,P. gingivalis）脂多糖（lipopolysaccharides,LPS）或热灭活P. gingivalis刺激HGFs,分别通过实时定量PCR 和ELISA观察EPA对白细胞介素6（interleukin-6,IL-6）、IL-8、IL-1β基因和蛋白表达的作用;进一步采用实时定量PCR 和Western免疫印迹法,观察EPA对血红素氧合酶1（heme oxygenase-1,HO-1）基因和蛋白表达的作用。采用SPSS 22.0软件包对数据进行统计学分析。结果: 200 μmol/L EPA可抑制HGFs的细胞活性;100 μmol/L EPA不影响HGFs的细胞活性、形态,对细胞周期也无显著影响（P>0.05）。EPA浓度依赖性抑制P. gingivalis LPS及热灭活P. gingivalis诱导HGFs表达的IL-6 mRNA、蛋白以及IL-1β mRNA（P<0.05）;EPA可呈浓度依赖性诱导HGFs表达HO-1 mRNA（P<0.05）,并上调其蛋白表达。结论: EPA在不影响HGFs生物学活性的前提下,显著抑制细胞的炎症因子表达,可能与其对HO-1的诱导作用有关,提示EPA在牙周炎防治中具有潜在应用价值。

关键词: 二十碳五烯酸, 牙龈成纤维细胞, 生物学活性, 炎症因子

Abstract: PURPOSE: To explore the effects of eicosapentaenoic acid (EPA) on biological activity and inflammatory factor expression of human gingival fibroblasts (HGFs). METHODS: The effects of EPA on the activity, morphology and cell cycle of HGFs were observed by living and dead cell staining, immunofluorescence staining and flow cytometry, respectively. HGFs were stimulated by lipopolysaccharides (LPS) of Porphyromonas gingivalis (P. gingivalis) or heat inactivated P. gingivalis, after which the effects of EPA on mRNA and protein expression of IL-6, IL-8 and IL-1β were observed by real-time PCR and ELISA, respectively. The gene and protein expression of heme oxygenase-1(HO-1) was also detected by real-time PCR and Western blotting, respectively. The data were analyzed with SPSS 22.0 software package. RESULTS: 200 μmol/L EPA inhibited cell activity of HGFs; 100 μmol/L EPA did not affect cell activity and morphology of HGFs, and had no significant effect on cell cycle (P>0.05). EPA significantly downregulated gene expression of IL-6 and IL-1β, and protein expression of IL-6 stimulated by P. gingivalis LPS and heat-killed P.gingivalis(P<0.05), in a dose-dependent manner. EPA increased gene expression of HO-1 in a dose dependent manner(P<0.05), and upregulated HO-1 protein expression. CONCLUSIONS: EPA significantly inhibits the expression of inflammatory factors without affecting the biological activity of HGFs, which may be related to the induction of HO-1, suggesting the potential role of EPA in the prevention and treatment of periodontitis.

Key words: Eicosapentaenoic acid, Human gingival fibroblasts, Biological activity, Inflammatory factor

中图分类号:

R781.4

周可聪, 孙梦君, 夏一如, 谢玉峰, 束蓉. 二十碳五烯酸对人牙龈成纤维细胞生物学活性及炎症因子表达的影响[J]. 上海口腔医学, 2021, 30(3): 225-231.

ZHOU Ke-cong, SUN Meng-jun, XIA Yi-ru, XIE Yu-feng, SHU Rong. Effects of eicosapentaenoic acid on biological activity and inflammatory factor expression of human gingival fibroblasts[J]. Shanghai Journal of Stomatology, 2021, 30(3): 225-231.

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二十碳五烯酸对人牙龈成纤维细胞生物学活性及炎症因子表达的影响

Effects of eicosapentaenoic acid on biological activity and inflammatory factor expression of human gingival fibroblasts

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