白藜芦醇对痛风患者外周血单个核细胞Sirtuin 1表达的影响

Effect of Sirtuin 1 expression in peripheral blood mononuclear cells from gout patients with resveratrol treatment

目的:白藜芦醇具有抗炎止痛、降尿酸等多种药理活性,但其机理尚未完全明确,白藜芦醇作为Sirtuin 1(SIRT1)的天然激动剂,本研究通过检测原发性痛风性关节炎患者外周血单个核细胞(PBMCs)中SIRT1 mRNA及蛋白表达水平,同时检测白藜芦醇体外干预尿酸盐晶体(MSU)刺激间歇期痛风患者PBMCs的SIRT1及IL-1β蛋白水平,探讨白藜芦醇干预痛风炎症的机理及意义。方法:收集原发性痛风性关节炎(GA)患者包括急性期(AG),间歇期(IG)和健康体检者(HC)的PBMCs,同时进行IG患者外周全血功能研究,予以不同浓度白藜芦醇干预MSU刺激IG患者6 h后分离PBMCs,采用实时荧光定量聚合酶链反应(RT-qPCR)检测SIRT1、NF-κB p65 mRNA表达水平和Western blot法检测SIRT1及IL-1β蛋白水平。结果:SIRT1 mRNA和蛋白水平在AG、IG组均低于HC(P分别<0. 01,0. 05);体外功能研究发现,200μmol/L和400μmol/L白藜芦醇干预MSU组的SIRT1蛋白水平与MSU组比较显著升高(P均<0. 01),而100μmol/L白藜芦醇干预MSU组SIRT1蛋白水平较MSU组有所升高,但无统计学意义; NF-κB p65 mRNA及IL-1β蛋白水平在200μmol/L和400μmol/L白藜芦醇干预MSU组较MSU组显著降低(P均<0. 01),100μmol/L白藜芦醇干预MSU组较MSU组无显著差异(P> 0. 05)。结论:痛风患者SIRT1表达水平降低,白藜芦醇通过浓度依赖性上调痛风患者SIRT1表达水平抑制致炎因子IL-1β产生,提示SIRT1是白藜芦醇抑制痛风炎症反应的作用靶点。

Objective： Resveratrol,which serves as the natural agonist of Sirtuin 1 （SIRT1）,has multiple pharmacological activity such as anti-inflammation,lowering uric acid,howerver,the mechanism of which is still elusive.SIRT1 mRNA and protein in peripheral blood mononuclear cells （PBMCs） from patients with primary gouty arthritis,SIRT1 and IL-1β proteins in PBMCs from monosodium urate（MSU）-stimulated intercritical gout patients with resveratrol treatment in vitro were studied to explore the pathogenesis of resveratrol involved in gouty inflammation. Methods： PBMCs from primary gouty arthritis patients including acute gout （AG）,intercritical gout （IG） as well as healthy control （HC） were obtained.In addition,IG patients were used to functional research in vitro,the PBMCs were obtained from MSU-stimulated IG patients with different doses of resveratrol for 6 h,SIRT1 and NF-κB p65 mRNA expression were detected by real-time quantification polymerase chain reaction （RT-qPCR） ,SIRT1 and IL-1β proteins were also measured using Western blot. Results： Both mRNA and proteins levels of SIRT1 were lower in patients with AG and IG groups than HC group （ P 〈0.01,0.05）.In the functional research in vitro,the SIRT1 protein increased in MSU-stimulated IG patients treated with resveratrol （200 μmol/L or 400 μmol/L） compared with MSU stimulation （ P 〈0.01）.However,the SIRT1 protein level in MSU treated with 100 μmol/L resveratrol was higher than that in MSU,but the difference was not statistically significant.What’s more,both NF-κB p65 mRNA and IL-1β protein levels decreased in MSU-stimulated IG patients treated with resveratrol （200 μmol/L or 400 μmol/L） compared with MSU stimulation （ P 〈0.01）,while there were similar IL-1β levels between resveratrol （100 μmol/L） and MSU stimulation （ P 〉0.05）. Conclusion： The expression of SIRT1 decreased in gout patients,and resveratrol inhibited pro-inflammatory cytokine IL-1β release in gout patients via up-regulation of SIRT1 dependent on concentration of resveratrol.It suggests that SIRT1 serves as a target of resveratrol reducing gouty inflammation.