白藜芦醇促进巨噬细胞M2极化并缓解小鼠急性痛风性关节炎

Resveratrol promotes macrophage M2 polarization and alleviates acute gouty arthritis in mice

目的观察白藜芦醇对小鼠急性痛风性关节炎的预防效果,并探讨巨噬细胞M2极化是否介导白藜芦醇的作用。方法18只C57BL/6小鼠随机分为3组,每组6只:假手术组、造模+溶剂对照组、造模+白藜芦醇组。假手术组用无菌生理盐水处理小鼠右侧后肢踝关节,造模+溶剂对照组小鼠右侧后肢踝关节给予单钠尿酸盐(MSU)晶体构建急性痛风性关节炎模型,造模+白藜芦醇组提前给予DMSO溶解的白藜芦醇干预再进行MSU晶体造模。测定各组小鼠双侧足掌厚度,采用H-E染色观察各组小鼠足掌关节滑膜组织炎症情况。分离、提取正常小鼠腹腔原代巨噬细胞,给予白藜芦醇干预后以MSU晶体刺激,用蛋白质印迹法检测巨噬细胞M1极化标志物诱导型一氧化氮合酶(iNOS)的表达,用实时荧光定量PCR检测巨噬细胞中炎症指标肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)mRNA的表达水平,用流式细胞术检测M2型巨噬细胞标志物F4/80、CD163的表达。结果成功构建小鼠急性痛风性关节炎模型。造模前给予白藜芦醇干预后,小鼠右侧后肢足掌厚度明显低于溶剂对照组[(1.98±0.02)mm vs(2.49±0.12)mm,P<0.01],小鼠足掌关节炎滑膜组织中中性粒细胞浸润面积减少。体外实验中,白藜芦醇显著抑制了小鼠腹腔原代巨噬细胞中iNOS蛋白和TNF-α、IL-1βmRNA的表达水平(P均<0.01),提高了F4/80^+CD163^+细胞比例(P<0.01)。结论白藜芦醇可能通过促进巨噬细胞M2极化,抑制炎性因子的产生,从而有效缓解小鼠急性痛风性关节炎。

Objective To investigate the preventive effect of resveratrol against acute gouty arthritis in mice and whether M2 polarization of macrophage mediates the effect.Methods Eighteen C57BL/6 mice were randomly divided into three groups(n=6):sham group,model+solvent control group,and model+resveratrol group.The right hind limb ankle joint of mice in the sham group were treated with sterile normal saline.The right hind limb ankle joint of mice in the model+solvent control group were treated with DMSO in advance and then with monosodium urate(MSU)crystals to establish acute gouty arthritis model.Mice in the model+resveratrol group were treated with resveratrol dissolved in DMSO in advance and then with MSU crystals to establish acute gouty arthritis model.The bilateral paw thickness of mice in each group was measured and H-E staining was used to observe the inflammation of synovial tissue of feet and metacarpal joints of mice in each group.The primary macrophages from abdominal cavity of normal mice were extracted,treated with resveratrol,and then stimulated with MSU crystals.The expression of M1-polarized macrophage markers inducible nitric oxide synthase(iNOS)protein and the inflammatory indexes tumor necrosis factorα(TNF-α)mRNA and interleukin 1β(IL-1β)mRNA were detected by Western blotting or qPCR.The expression of M2-polarized macrophage markers F4/80 and CD163 were detected by flow cytometry.Results Acute gouty arthritis model of mice was successfully established.The right hind limb thickness of mice in the model+resveratrol group was significantly lower than that in the model+solvent control group([1.98±0.02]mm vs[2.49±0.12]mm,P<0.01).The infiltration area of neutrophils in synovial tissue of feet and metacarpal joints in mice of model+resveratrol group were also significantly reduced.In vitro,resveratrol significantly inhibited the expression of iNOS protein,TNF-αmRNA and IL-1βmRNA in primary peritoneal macrophages(all P<0.01)and increased the percentage of F4/80^+CD163^+in macrophages(P<0.01).Conclusion Resveratrol may effectively alleviate acute gouty arthritis in mice by promoting M2 polarization of macrophages and inhibiting the expression of inflammatory factors.