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

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 20 μmol/L 组、RSV 40 μmol/L组、RSV 80 μmol/L组、LPS+RSV 20 μmol/L组、LPS+RSV 40 μmol/L组和LPS+RSV 80 μmol/L组；实 验2细胞分为对照组、LPS组、LPS+RSV 40 μmol/L 组、LPS+RSV 40 μmol/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80 μmol/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80 μmol/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，80 μmol/L RSV显著提高了GSH-Px水平（P＜0.05）。此外，20、40和80 μmol/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。

Resveratrol prevents inflammation and oxidative stress response in HGFs

Objective To elucidate the anti-inflammatory and anti-oxidative properties of resveratrol (RSV) on Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide- (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+RSV 40 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.