白藜芦醇抑制人牙龈成纤维细胞炎症和氧化应激的机制研究  

Resveratrol prevents inflammation and oxidative stress response in HGFs

作　　者：李丽华[1] 王玉娇 李俊雄 李思玉 唐婉容[1] 邱亚[1] LI Li-hua;WANG Yu-jiao;LI Jun-xiong;LI Si-yu;TANG Wan-rong;QIU Ya(Department of Stomatology,the Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院口腔科,637000

出　　处：《天津医药》2021年第12期1245-1249,共5页Tianjin Medical Journal

基　　金：南充市校战略合作科技专项资金(18SXHZ0092);南充市校战略合作科技专项资金(19SXHZ0078);南充市科学技术局科技计划项目(20YFZJ0090)。

摘　　要：目的探讨白藜芦醇(RSV)对牙龈卟啉单胞菌脂多糖(LPS)诱导的人牙龈成纤维细胞(HGF)的炎症和氧化应激的调节作用。方法原代培养HGF,将细胞分为实验1和实验2:实验1细胞分为对照组、LPS组、RSV 20μmol/L组、RSV 40μmol/L组、RSV 80μmol/L组、LPS+RSV 20μmol/L组、LPS+RSV 40μmol/L组和LPS+RSV 80μmol/L组;实验2细胞分为对照组、LPS组、LPS+RSV 40μmol/L组、LPS+RSV 40μmol/L+E5564组和LPS+E5564组。通过CCK-8法评估细胞活力。利用酶联免疫吸附试验测定白细胞介素(IL)-1β、IL-6、IL-8、肿瘤坏死因子(TNF)-α、超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)水平。通过Western blot分析测量蛋白的表达水平。结果20、40和80μmol/L RSV对HGF均没有明显的细胞毒性作用。在LPS诱导的HGF细胞中,40和80μmol/L RSV通过下调IL-1β、IL-6、IL-8和TNF-α的表达减弱炎症反应,降低了MDA的含量,并显著提高了SOD的水平,80μmol/L RSV显著提高了GSH-Px水平(P<0.05)。此外,20、40和80μmol/L RSV可诱导Toll样受体4(TLR4)/MyD88/NF-κB信号通路的失活,其均可降低TLR4、MyD88和p-p65蛋白的表达水平(P<0.05)。TLR4抑制剂(E5564)通过下调IL-1β、IL-6、IL-8和TNF-α的产生以及上调GSH-Px水平进一步增强了RSV对炎症和氧化应激损伤的缓解作用(P<0.05)。结论 RSV可通过诱导TLR4/MyD88/NF-κB信号通路失活,减轻LPS导致的HGF炎症和氧化应激损伤。Objective To elucidate the anti-inflammatory and anti-oxidative properties of resveratrol(RSV) on Porphyromonas gingivalis(P.gingivalis) lipopoly saccharide-(LPS-) induced human gingival fibroblasts(HGFs).Methods HGF cells were divided into experiment 1 and experiment 2.Cells in experiment 1 were divided into the control group,the LPS group,the RSV 20 μmol/L group,the RSV 40 μmol/L group,the RSV 80 μmol/L group and the LPS+RSV 20 μmol/L group.The cells in experiment 2 were divided into the control group,the LPS group,the LPS+RSV 40 μmol/L group,the LPS+RSV40 μmol/L+E5564 group and the LPS+E5564 group.Cell viability was evaluated by cell-counting kit-8 assay.IL-1β,IL-6,IL-8,TNF-α,SOD,MDA,and GSH-Px levels were measured by ELISA.Protein expressions were measured by Western blot analysis.Results It was found that 20,40 and 80 μmol/L RSV had no significant effects on the viability of HGF cells.In LPSinduced HGFs,40 and 80 μmol/L RSV significantly reduced inflammation by the down-regulation of IL-1β,IL-6,IL-8,and TNF-α expression.And 40 and 80 μmol/L RSV also decreased MDA expression,accompanied by an increase of SOD production(P<0.05).Meanwhile,80 μmol/L RSV significantly increased GSH-Px levels(P<0.05).Additionally,20,40 and 80 μmol/L RSV induced deactivation of TLR4/MyD88/NF-κB signaling pathway(P<0.05).It was found that TLR4 inhibitor(E5564) could further strengthen RSV-reduced inflammation and OS injury by the down-regulation of IL-1β,IL-6,IL-8,and TNF-α production and up-regulation of GSH-Px in LPS-induced HGFs(P<0.05).Conclusion Resveratrol attenuates the inflammation and OS injury of P.gingivalis LPS-treated HGFs by deactivating TLR4/MyD88/NF-κB signaling pathway.

关 键 词：藜芦 醇类 信号传导 NF-κB 慢性牙周炎 成纤维细胞 白藜芦醇 

分 类 号：R781.42[医药卫生—口腔医学]