冯佳, 黄侠, 李赫宇, 鞠单, 杨年安, 田瑞, 夏燕, 袁林. 白藜芦醇抑制单钠尿酸盐诱导RAW264.7巨噬细胞氧化损伤的机制J. 药学学报, 2020,55(10): 2368-2374. doi: 10.16438/j.0513-4870.2020-0065
引用本文: 冯佳, 黄侠, 李赫宇, 鞠单, 杨年安, 田瑞, 夏燕, 袁林. 白藜芦醇抑制单钠尿酸盐诱导RAW264.7巨噬细胞氧化损伤的机制J. 药学学报, 2020,55(10): 2368-2374. doi: 10.16438/j.0513-4870.2020-0065
FENG Jia, HUANG Xia, LI He-yu, JU Dan, YANG Nian-an, TIAN Rui, XIA Yan, YUAN Lin. Mechanism of resveratrol inhibiting monosodium urate induced oxidative damage of RAW264.7 macrophagesJ. Acta Pharmaceutica Sinica, 2020,55(10): 2368-2374. doi: 10.16438/j.0513-4870.2020-0065
Citation: FENG Jia, HUANG Xia, LI He-yu, JU Dan, YANG Nian-an, TIAN Rui, XIA Yan, YUAN Lin. Mechanism of resveratrol inhibiting monosodium urate induced oxidative damage of RAW264.7 macrophagesJ. Acta Pharmaceutica Sinica, 2020,55(10): 2368-2374. doi: 10.16438/j.0513-4870.2020-0065

白藜芦醇抑制单钠尿酸盐诱导RAW264.7巨噬细胞氧化损伤的机制

Mechanism of resveratrol inhibiting monosodium urate induced oxidative damage of RAW264.7 macrophages

  • 摘要: 探讨白藜芦醇对单钠尿酸盐(monosodium urate,MSU)晶体诱导小鼠RAW264.7巨噬细胞的炎症因子、氧化应激指标及核因子E2相关因子2(nuclear factor erythroid-2-related factor 2,Nrf2)/血红素氧合酶1(heme oxygenase 1,HO-1)信号通路相关基因的作用机制,为急性痛风性关节炎(acute gouty arthritis,AGA)的治疗提供理论依据。使用不同浓度白藜芦醇作用于RAW264.7细胞5 h后,再加入MSU刺激24 h。采用CCK-8法检测白藜芦醇对RAW264.7细胞增殖的作用;ELISA法检测细胞分泌肿瘤坏死因子α(tumour necrosis factor-α, TNF-α)水平;应用2',7'-二氯荧光素二乙酸酯(2',7'-dichlorodi-hydrofluorescein diacetate,DCFH-DA)探针法检测细胞内活性氧自由基(reaction oxygen species,ROS);测定细胞内超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malonaldehyde,MDA)含量;实时荧光定量PCR(real-time PCR)法检测Nrf2、Kelch样ECH相关蛋白1(Kelch-like ECH-associated protein 1,Keap1)、醌氧化还原酶1NAD(P)H quinine oxidoreductase 1,NQO1和HO-1 mRNA的表达。结果显示,白藜芦醇能够抑制RAW264.7细胞增殖;明显抑制MSU诱导RAW264.7细胞分泌的TNF-α水平;明显抑制MSU诱导RAW264.7细胞内的ROS、MDA表达,增加SOD表达;降低MSU诱导RAW264.7细胞内的Keap1 mRNA表达,增高Nrf2、NQO1和HO-1 mRNA的表达。综上,白藜芦醇能够抑制MSU诱导RAW264.7巨噬细胞产生的炎症反应,并通过调节Nrf2/HO-1信号通路提高巨噬细胞抗氧化能力。

     

    Abstract: The aim of this research is to investigate the effects of resveratrol on the inflammatory factors, oxidative stress indexes and the related genes of nuclear factor erythroid-2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in RAW264.7 macrophages induced by monosodium urate (MSU) and to provide a theoretical basis for the treatment of acute gouty arthritis (AGA). Different concentrations of resveratrol were used to treat RAW264.7 cells for 5 hours, then MSU was added to stimulate them for 24 hours. The proliferation of RAW264.7 cells were detected by CCK-8 method. The level of tumor necrosis factor α secreted by cells were detected by ELISA method. The content of reaction oxygen species (ROS) in cells were detected by 2',7'-dichlorodi-hydrofluorescein diacetate (DCFH-DA) probe method. The contents of superoxide dismutase (SOD) and malonaldehyde (MDA) in cells were detected by the kits. The expression of Nrf2, Kelch like ECH related protein 1 (Keap1), NAD(P)H quinone oxidoreductase 1 (NQO1), HO-1 mRNA were detected by real time PCR method. The results showed that resveratrol could inhibit the proliferation of RAW264.7 cells, significantly inhibit the TNF-α level of RAW264.7 cells induced by MSU, significantly inhibit the expression of ROS and MDA, and increase the expression of SOD in RAW264.7 cells induced by MSU. Resveratrol could reduce the expression of Keap1 mRNA in RAW264.7 cells induced by MSU, and increase the expression of Nrf2, NQO1, HO-1 mRNA. It is suggested that resveratrol can inhibit the inflammatory response of RAW264.7 macrophages induced by MSU, and improve the antioxidant capacity of macrophages by regulating Nrf2/HO-1 signal pathway.

     

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