摘要
目的探索二十二碳六烯酸(docosahexaenoic acid, DHA)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作用。方法分别采用活死细胞染色法、荧光染色法、流式细胞术观察DHA对细胞活性、细胞形态、细胞周期的影响;DHA预处理HGFs后,利用牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)脂多糖(lipopolysaccharides, LPS)或热灭活P.gingivalis刺激细胞,定量PCR和ELISA观察白介素-6(interleukin-6,IL-6)、IL-8、IL-1β基因和蛋白表达的变化;通过定量PCR和Western blot检测DHA对血红素氧合酶-1(heme oxygenase-1,HO-1)基因和蛋白表达的作用。结果 200μmol/L DHA可降低HGFs的细胞活性,但100μmol/L DHA不影响细胞活性及形态,对细胞周期也无明显影响(P>0.05);DHA可抑制P.gingivalis LPS或热灭活P.gingivalis诱导HGFs表达的IL-6、IL-8、IL-1βmRNA以及IL-6、IL-8蛋白(P<0.05),并可显著上调HGFs表达的HO-1 mRNA及蛋白(P<0.05)。结论 DHA可显著抑制HGFs的炎症因子表达,并且不影响其生物学活性,其抗炎作用可能与HO-1上调相关。
Objective To explore the effects of docosahexaenoic acid(DHA) on the biological activity and inflammatory response of human gingival fibroblasts(HGFs). Methods The effects of DHA on cell viability, cell morphology and cell cycle were observed by living and dead cell staining, fluorescent staining and flow cytometry, respectively. After pretreated with DHA, HGFs were stimulated with lipopolysaccharide(LPS) from Porphyromonas gingivalis(P.gingivalis) or with heat-inactivated P.gingivalis. Then the gene and protein expressions of interleukin-6(IL-6), IL-8 and IL-1β were detected by quantitative PCR and ELISA, respectively. The effects of DHA on heme oxygenase-1(HO-1) gene and protein expression were also observed by quantitative PCR and Western blot, respectively. Results The 200 μmol/L DHA reduced the cell activity of HGFs, but 100 μmol/L DHA had no effect on cell viability and morphology, and had no significant effect on cell cycle(P>0.05). DHA inhibited the expression of IL-6, IL-8, IL-1β mRNA and IL-6, IL-8 protein induced by P.gingivalis LPS or heat-inactivated P.gingivalis(P<0.05). DHA significantly induced HO-1 mRNA and protein expression(P<0.05). Conclusion DHA can significantly inhibit the inflammatory response of HGFs without affecting the biological activity. The anti-inflammatory effect may be related to the induction of HO-1.
作者
孙梦君
周可聪
夏一如
谢玉峰
束蓉
SUN Mengjun;ZHOU Kecong;XIA Yiru;XIE Yufeng;SHU Rong(Depurment of Periodontology,Shanghai Ninth Peoples Hospial,Shanghai Jiao Tong University School of Medicine,College of Stomatology,Shanghai Jiao Tong University,National Center for Stomalology,National Clinical Research Center for Oral Diseases,Shanghai Key Laboratory of Stomatology,Shanghai 200011,China)
出处
《口腔医学》
CAS
2021年第5期385-390,共6页
Stomatology
基金
国家自然科学基金重大项目(81991500,81991503)
口腔医学重点核心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)
九院基础研究助推计划(JYZZ108)。
