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.

参考文献

[1] Hajishengallis G, Chavakis T, Lambris JD.Current understanding of periodontal disease pathogenesis and targets for host-modulation therapy[J]. Periodontol 2000, 2020, 84(1): 14-34.
[2] Scannapieco FA, Gershovich E.The prevention of periodontal disease-an overview[J]. Periodontol 2000, 2020, 84(1): 9-13.
[3] Chopra A, Bhat SG, Sivaraman K.Porphyromonas gingivalis adopts intricate and unique molecular mechanisms to survive and persist within the host: a critical update[J]. J Oral Microbiol, 2020, 12(1): 1801090.
[4] Suzumura A, Terao R, Kaneko H.Protective effects and molecular signaling of n-3 fatty acids on oxidative stress and inflammation in retinal diseases[J]. Antioxidants (Basel), 2020, 9(10): 920-928.
[5] Oppedisano F, Macrì R, Gliozzi M, et al.The anti-inflammatory and antioxidant properties of n-3 PUFAs: their role in cardiovascular protection[J]. Biomedicines, 2020, 8(9): 306-325.
[6] Sun M, Zhou Z, Dong J, et al.Antibacterial and antibiofilm activities of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) against periodontopathic bacteria[J]. Microb Pathog, 2016, 99: 196-203.
[7] 周子超. DHA和EPA对P.gingivalis LPS诱导人牙周膜成纤维细胞炎症反应的影响 [D]. 上海: 上海交通大学, 2015.
[8] Sheng H, Li P, Chen X, et al.Omega-3 PUFAs induce apoptosis of gastric cancer cells via ADORA1[J]. Front Biosci (Landmark Ed), 2014, 19: 854-861.
[9] Eser PO, Vanden Heuvel JP, Araujo J, et al.Marine- and plant-derived ω-3 fatty acids differentially regulate prostate cancer cell proliferation[J]. Mol Clin Oncol, 2013, 1(3): 444-452.
[10] Meng H, Shen Y, Shen J, et al.Effect of n-3 and n-6 unsaturated fatty acids on prostate cancer(PC-3) and prostate epithelial (RWPE-1) cells in vitro[J]. Lipids Health Dis, 2013, 12: 160-174.
[11] Xue M, Wang Q, Zhao J, et al.Docosahexaenoic acid inhibited the Wnt/β-catenin pathway and suppressed breast cancer cells in vitro and in vivo [J]. J Nutr Biochem, 2014, 25(2): 104-110.
[12] Zajdel A, Wilczok A, Chodurek E, et al.Polyunsaturated fatty acids inhibit melanoma cell growth in vitro[J]. Acta Pol Pharm, 2013, 70(2): 365-369.
[13] Peng Y, Zheng Y, Zhang Y, et al.Different effects of omega-3 fatty acids on the cell cycle in C2C12 myoblast proliferation[J]. Mol Cell Biochem, 2012,367(1-2): 165-173.
[14] Barascu A, Besson P, Le Floch O, et al.CDK1-cyclin B1 mediates the inhibition of proliferation induced by omega-3 fatty acids in MDA-MB-231 breast cancer cells[J]. Int J Biochem Cell Biol, 2006, 38(2): 196-208.
[15] Li Q, Cui K, Wu M, et al.Polyunsaturated fatty acids influence LPS-induced inflammation of fish macrophages through differential modulation of pathogen recognition and p38 MAPK/NF-κB signaling[J]. Front Immunol, 2020, 11: 559332.
[16] Abekura Y, Ono I, Kawashima A, et al.Eicosapentaenoic acid prevents the progression of intracranial aneurysms in rats[J]. J Neuroinflammation, 2020, 17(1): 129-144.
[17] Oliver E, McGillicuddy FC, Harford KA, et al. Docosahexaenoic acid attenuates macrophage-induced inflammation and improves insulin sensitivity in adipocytes-specific differential effects between LC n-3 PUFA[J]. J Nutr Biochem, 2012, 23(9): 1192-1200.
[18] Martinez-Micaelo N, González-Abuín N, Terra X, et al.Omega-3 docosahexaenoic acid and procyanidins inhibit cyclo-oxygenase activity and attenuate NF-κB activation through a p105/p50 regulatory mechanism in macrophage inflammation[J]. Biochem J, 2012, 441(2): 653-663.
[19] Wang Y, He J, Yang J.Eicosapentaenoic acid improves polycystic ovary syndrome in rats via sterol regulatory element-binding protein 1 (SREBP-1)/toll-like receptor 4 (TLR4) pathway[J]. Med Sci Monit, 2018, 24: 2091-2097.
[20] Solís-Martínez O, Plasa-Carvalho V, Phillips-Sixtos G, et al.Effect of eicosapentaenoic acid on body composition and inflammation markers in patients with head and neck squamous cell cancer from a public hospital in Mexico[J]. Nutr Cancer, 2018, 70(4): 663-670.
[21] Wang H, Khor TO, Saw CL, et al.Role of Nrf2 in suppressing LPS-induced inflammation in mouse peritoneal macrophages by polyunsaturated fatty acids docosahexaenoic acid and eicosapentaenoic acid[J]. Mol Pharm, 2010, 7(6): 2185-2193.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	周子超, 束蓉, 吴轶群. EPA对P.gingivalis LPS诱导人牙周膜成纤维细胞炎症反应的影响[J]. 上海口腔医学, 2023, 32(5): 475-479.
[2]	张世坤, 裴冬冬, 姜丹丹, 张丽华, 蔡建英, 常忠福. 甲硝唑联合米诺环素在局限性牙周炎种植牙术后感染中的应用效果评价[J]. 上海口腔医学, 2023, 32(5): 532-535.
[3]	万志群, 周正根, 王竞, 周群. 微小RNA-100-5p对颞下颌关节炎中雷帕霉素靶蛋白的作用机制研究[J]. 上海口腔医学, 2022, 31(1): 12-16.
[4]	许辛夷, 张银莲. TiO₂-PEEK-CRGDS材料制备及抗牙龈卟啉单胞菌活性和细胞活性分析[J]. 上海口腔医学, 2022, 31(1): 54-57.
[5]	牛晨光, 李佳洋, 於丽明, 陈栋, 韦晓玲. 牡荆素对脂多糖诱导牙髓干细胞表达炎症因子的影响[J]. 上海口腔医学, 2020, 29(5): 462-465.
[6]	王震, 孙扬, 陈佳露, 龚逸明. 姜黄素对环孢素A诱导激活的大鼠牙龈成纤维细胞TGF-β1/Smad3通路的抑制作用[J]. 上海口腔医学, 2020, 29(4): 359-364.
[7]	张佳丽, 姚军, 诸葛继娜, 张琰君. 赤藓糖醇对牙周致病菌抗菌性能的影响[J]. 上海口腔医学, 2019, 28(4): 362-367.
[8]	赖欣添, 夏一如, 谢玉峰, 束蓉. 外源性ATP对牙龈成纤维细胞NLRP3炎症小体活化的影响[J]. 上海口腔医学, 2018, 27(4): 342-348.
[9]	张长源, 牛刚, 靳祎祎, 于皓, 程辉. 3种树脂水门汀对人牙龈成纤维细胞的体外细胞毒性[J]. 上海口腔医学, 2017, 26(6): 594-598.
[10]	邵珺雅, 汪建中, 徐梦蓉, 周兰芳. 烟草对人牙龈成纤维细胞增殖及纤维连接蛋白表达的影响[J]. 上海口腔医学, 2017, 26(1): 7-11.
[11]	韦晓玲, 刘道杨, 潘杰, 陆珮珺, 赵君. 冻干对BMP-2基因重组慢病毒载体生物学效应的影响[J]. 上海口腔医学, 2016, 25(5): 522-526.
[12]	呼海燕, 高志彪. Egr-1对机械力诱导人牙周膜细胞炎症反应的作用及机制研究[J]. 上海口腔医学, 2016, 25(5): 553-559.
[13]	李欣, 洪弘, 张延清, 韦曦. 大鼠牙乳头细胞旁分泌效应对巨噬细胞分泌炎症因子的调控作用[J]. 上海口腔医学, 2016, 25(4): 396-402.
[14]	孟令娇，仇丽鸿，于雅琼，郭佳杰，詹福良，张凌，张晓芳. 氢氧化钙对根尖周成骨细胞IL-6、TNF-α表达的影响[J]. 上海口腔医学, 2016, 25(1): 32-37.
[15]	孙志新,张云涛. IL-1β对人牙龈成纤维细胞在不同钛板表面表达炎性因子的影响[J]. 上海口腔医学, 2015, 24(2): 151-156.