白藜芦醇对类风湿性关节炎患者成纤维样滑膜细胞cGAS-STING信号通路的影响  

Effects of resveratrol on cGAS-STING signaling pathway in fibroblast-like synoviocytes of patients with rheumatoid arthritis

作　　者：汪陶荣 邵玉宝 刘楠楠 李文昊 李梦 陈晓宇[1,2] Wang Taorong;Shao Yubao;Liu Nannan;Li Wenhao;Li Meng;Chen Xiaoyu(Microscopic Morphological Center Laboratory,Anhui Medical University,Hefei 230032;Dept of Histology and Embryology,Anhui Medical University,Hefei 230032;Dept of Respiratory and Critical Care Medicine,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Pharmacy,Bozhou Vocational and Technical College,Bozhou 236800)

机构地区：[1]安徽医科大学形态学中心实验室,合肥230032 [2]安徽医科大学组织与胚胎学教研室,合肥230032 [3]安徽医科大学第一附属医院呼吸与危重症医学科,合肥230022 [4]亳州职业技术学院药学系,亳州236800

出　　处：《安徽医科大学学报》2025年第1期73-78,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高校科研计划项目(编号:2023AH050607);安徽医科大学校科研基金(编号:2021xkj004)。

摘　　要：目的探究白藜芦醇(Res)对类风湿性关节炎(RA)患者成纤维样滑膜细胞(FLS)的影响,并初步分析Res抑制FLS释放炎症因子的可能机制。方法体外培养RA患者FLS细胞,采用不同浓度的Res(0、20、40、80、160、320μmol/L)进行处理,12、24 h后CCK-8法分别检测FLS细胞活力;采用不同浓度的Res(0、40、80、160μmol/L)进行处理FLS细胞24 h后,ELISA法分析FLS细胞分泌的炎症因子白细胞介素(IL)-6和肿瘤坏死因子-α(TNF-α)水平,Western blot测定细胞中环磷酸鸟苷-腺苷酸合成酶(cGAS)、干扰素刺激因子(STING)蛋白的表达水平;慢病毒感染FLS使其过表达cGAS,将细胞分为Control(对照)组、cGAS组(cGAS过表达)、Res+cGAS组(Res 160μmol/L+cGAS过表达)、Res组(Res 160μmol/L)。Western blot测定各组细胞中STING蛋白的表达水平,CCK-8法检测各组FLS细胞活力,ELISA法检测各组细胞上清液中炎症因子IL-6和TNF-α含量。结果CCK-8实验结果显示,与0μmol/L Res组比较,使用40、80、160μmol/L的Res处理24 h后的三组FLS细胞活力下降(P<0.01);ELISA结果也表明与0μmol/L Res组比较,使用40、80、160μmol/L的Res处理后的三组FLS细胞上清液中IL-6和TNF-α含量也降低(P<0.01)。同时,Western blot结果显示,与0μnmol/L的Res组相比,经过40、80、160μnmol/L的Res处理后的三组FLS细胞中STING和cGAS蛋白表达量降低,差异有统计学意义(P<0.05,P<0.01)。与Control组相比,过表达cGAS后,CGAS组中FLS中STING蛋白表达水平增加(P<0.05),与Res组相比,Res+cGAS组中FLS细胞上清液中炎症因子含量及FLS中STING蛋白表达水平显著增加(P<0.01,P<0.05)。结论适当剂量的Res能有效减少FLS细胞分泌炎症因子,而这一抑制作用机制可能源于其对cGAS-STING信号传导途径的阻断作用。Objective To investigate the effects of resveratrol(Res)on fibroblast-like synoviocytes(FLS)in patients with rheumatoid arthritis(RA),and to explore the possible mechanism of Res inhibiting the release of inflammatory factors from FLS.Methods FLS from RA patients were cultured in vitro and treated with different concentrations of Res(0,20,40,80,160,320μmol/L).The viability of FLS cells was detected by CCK-8 assay after 12 and 24 h.The contents of inflammatory factor interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in cell supernatant were detected by ELISA.The expression levels of cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)and stimulator of interferon gene(STING)were measured by Western blot;After lentivirus infection with FLS caused the cells to overexpress cGAS,the cells were divided into Control group(blank control),cGAS group(cGAS overexpression),Res+cGAS group(Res 160μmol/L+cGAS overexpression)and Res group(Res 160μmol/L).The expression level of STING protein in cells of each group was determined by Western blot,the viability of FLS cells in each group was detected by CCK-8,and the contents of inflammatory factor IL-6 and TNF-αin the supernatant of cells of each group were detected by ELISA method.Results The results of CCK-8 experiment showed that under 40,80,160μmol/L Res treatment,FLS viability decreased significantly after 24 h compared with blank control group(P<0.01).ELISA results showed that the contents of IL-6 and TNF-αin cell supernatant were also significantly decreased after treatment with Res of 40,80 and 160μmol/L(P<0.01).Meanwhile,Western blot results showed that Res could significantly decrease the protein expression levels of STING and cGAS in FLS cells after treatment of 40,80 and 160μmol/L(P<0.05,P<0.01).Compared with the Control group,the expression level of STING protein in FLS increased after overexpression of cGAS(P<0.05);compared with the Res group,the content of inflammatory factors in the supernatant of FLS and the expression level of STING pr

关 键 词：类风湿性关节炎 成纤维样滑膜细胞 白藜芦醇 cGAS-STING信号通路 白介素-6 肿瘤坏死因子-α 

分 类 号：R285[医药卫生—中药学]