二苯乙烯苷抗氧化和抗炎作用的机制研究

doi:10.3969/j.issn.1004-616x.2020.02.005

癌变·畸变·突变 ›› 2020, Vol. 32 ›› Issue (2): 105-111,117.doi: 10.3969/j.issn.1004-616x.2020.02.005

二苯乙烯苷抗氧化和抗炎作用的机制研究

陈子卓¹, 徐宇航¹, 赵九洲¹, 朱敬朴¹, 郑义鹏¹, 李文丽², 吴浩芝¹, 海春旭², 于卫华²

1. 空军军医大学基础医学院, 陕西西安 710032;
2. 空军军医大学军事预防医学院军事毒理学与防化医学教研室, 陕西省自由基生物学与医学重点实验室, 特殊作业环境危害评估与防治教育部重点实验室. 陕西西安 710032

收稿日期:2019-12-18 修回日期:2020-02-11 出版日期:2020-03-30 发布日期:2020-04-10
通讯作者: 海春旭,E-mail:cx-hai@fmmu.edu.cn;于卫华,E-mail:yuweihua_fmmu@163.com E-mail:cx-hai@fmmu.edu.cn;yuweihua_fmmu@163.com
作者简介:陈子卓,E-mail:761795560@qq.com;徐宇航,E-mail:635427714@qq.com;赵九洲,E-mail:18802972515@163.com。
基金资助:
国家自然科学基金青年科学基金（31800706）；国家自然科学基金面上项目（21677176，81473010）

Antioxidant and anti-inflammatory effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in macrophages

CHEN Zizhuo¹, XU Yuhang¹, ZHAO Jiuzhou¹, ZHU Jingpu¹, ZHENG Yipeng¹, LI Wenli², WU Haozhi¹, HAI Chunxu², YU Weihua²

1. School of Basic Medical Sciences, Air Force Medical University, Xi'an 710032;
2. Department of Toxicology, Key Lab of Hazard Assessment and Control in Special Operational Environment of Ministry of Education, Shaanxi Provincial Key Lab of Free Radical Biology and Medicine, School of Public Health, Air Force Medical University, Xi'an 710032, Shaanxi, China

Received:2019-12-18 Revised:2020-02-11 Online:2020-03-30 Published:2020-04-10

摘要/Abstract

摘要： 目的： 以小鼠巨噬细胞RAW264.7细胞为研究对象，探讨二苯乙烯苷（TSG）介导的抗氧化和抗炎作用及其调控机制。方法： 30、60和120 μmol/L的二苯乙烯苷预处理4 h，然后以1 μg/mL的脂多糖（LPS）刺激小鼠巨噬细胞（RAW264.7）12 h，并设置空白对照组、地塞米松（100 nmol/L）阳性对照和Nrf2抑制剂寒鸦子酸（brusato，50 μmol/L）干预组，其中地塞米松和brusatol预处理RAW264.7细胞4 h。观察光镜下RAW264.7细胞形态学的改变并统计其被激活比例；ELISA法检测培养液中TNF-α和IL-6的水平；通过DCFH-DA和Mito-SOX^TM荧光探针染色，流式细胞仪检测细胞内和线粒体内的ROS水平变化；免疫荧光法检测Nrf2的表达水平变化；Western blot检测核内Nrf2蛋白水平变化；Real-time PCR测定Nrf2下游抗氧化酶HO-1、SOD₂、SOD₁、CAT和GPX-1表达。结果： 与空白对照组细胞相比，LPS处理后RAW264.7细胞活化，胞体增大，形成大量伪足，培养基中TNF-α和IL-6增多。二苯乙烯苷可抑制LPS诱导的RAW264.7细胞激活，降低培养基中TNF-α和IL-6，且效果优于阳性对照药物地塞米松。同时，二苯乙烯苷可促进Nrf2核转位，并促进下游抗氧化酶HO-1、SOD₂和CAT表达，减轻细胞内ROS生成。而brusatol可通过阻断Nrf2活性抑制二苯乙烯苷的抗炎作用。结论： 二苯乙烯苷可激活Nrf2及其下游抗氧化酶的表达，具有良好的抗炎和抗氧化作用，是一种炎症相关疾病的候选药物。

关键词: 二苯乙烯苷, 抗氧化, 抗炎, 核因子E2相关因子2, RAW264.7细胞

Abstract: OBJECTIVE: To explore the induction of antioxidant and anti-inflammation effects of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) in LPS-stimulated RAW264.7 macrophages. METHODS: RAW264.7 cells were pretreated with TSG (30,60,120 μmol/L) for 4 hours,and then exposed to 1 μg/mL LPS for 12 hours. In addition,a blank control and two positive control groups were set up. For the positive controls,cells were treated with dexamethasone (100 nmol/L) or brusatol (50 μmol/L) at 4 h prior to LPS stimulation. Morphological changes were captured under a microscope,and the activated cell rate was analyzed. Concentrations of TNF-α and IL-6 in the culture medium were assessed using specific ELISA kits. Levels of superoxide anions and reactive oxygen species within the cells were detected using flow-cytometry after staining with MitoSOX^TM and DCFH-DA,respectively. Moreover,immunofluorescence staining and western-blot methods were used to investigate protein expression and sub-cellular localization of Nrf2 in the cells. Real-time PCR was used to detect expression of Nrf2-related antioxidant enzymes,including HO-1,SOD₂,SOD₁,CAT and GPX-1. RESULTS: Compared with the control group,LPS activated macrophages exhibited elongated cell morphology with pseudopodium-like protrusions. LPS also strongly promoted the level of pro-inflammatory cytokines,including TNF-α and IL-6. However,pretreatment with TSG inhibited LPS-induced ROS production and inflammatory cytokines secretion. Interestingly,the anti-inflammatory effect of TSG was better than Dex. TSG administration promoted Nrf2 protein expression and its translocated into the nucleus. Moreover,TSG alleviated LPS-induced oxidative stress through regulating Nrf2-related antioxidant enzymes,including HO-1,SOD₂ and CAT. Brusatol,a specific inhibitor of Nrf2,decreased TNF-α and IL-6 production in LPS-activated macrophages. CONCLUSION: TSG attenuated LPS-induced oxidative stress and inflammation in RAW264.7 macrophages through activation of Nrf2 signal pathway. Therefore,TSG may be useful as a drug to prevent or to treat inflammation-associated diseases.

Key words: 2, 3, 5, 4'-tetrahydroxystilbene 2-O-β-D-glucoside, antioxidant, anti-inflammation, Nrf2, RAW264.7 cells

中图分类号:

R114

陈子卓, 徐宇航, 赵九洲, 朱敬朴, 郑义鹏, 李文丽, 吴浩芝, 海春旭, 于卫华. 二苯乙烯苷抗氧化和抗炎作用的机制研究[J]. 癌变·畸变·突变, 2020, 32(2): 105-111,117.

CHEN Zizhuo, XU Yuhang, ZHAO Jiuzhou, ZHU Jingpu, ZHENG Yipeng, LI Wenli, WU Haozhi, HAI Chunxu, YU Weihua. Antioxidant and anti-inflammatory effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in macrophages[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(2): 105-111,117.

参考文献

[1] 李文军, 李二红, 杨宗城. 巨噬细胞在炎症消散中的作用[J]. 免疫学杂志, 2000, 16(Sup 1):99-101.
[2] OISHI Y, MANABE I. Macrophages in inflammation, repair and regeneration[J]. Int Immunol, 2018, 30(11):511-528.
[3] HOU W, HU S Y, SU Z Z, et al. Myricetin attenuates LPS-induced inflammation in RAW 264.7 macrophages and mouse models[J]. Future Med Chem, 2018, 10(19):2253-2264.
[4] SINGLA B, HOLMDAHL R, CSANYI G. Editorial:oxidants and redox signaling in inflammation[J]. Front Immunol, 2019, 10:545.
[5] 于卫华, 周庆彪, 刘颖, 等. 活性氧调控炎症诱发肿瘤机制的研究进展[J]. 癌变·畸变·突变, 2016, 28(2):158-160.
[6] BISWAS S K. Does the interdependence between oxidative stress and inflammation explain the antioxidant paradox?[J]. Oxid Med Cell Longev, 2016, 2016:5698931.
[7] 李越, 沈磊, 罗和生. Nrf2/ARE通路在炎症性肠病中的研究进展[J]. 胃肠病学和肝病学杂志, 2016, 25(7):721-723.
[8] PASTOREK M, MÜLLER P, VOJTĚŠEK B. Nrf2:two faces of antioxidant system regulation[J]. Klin Onkol, 2015, 28(Suppl 2):2S26-2S31.
[9] 黄世琼, 张毅, 杨军宣. 何首乌主要成分二苯乙烯苷的研究进展[J]. 海峡药学, 2016, 28(6):37-39.
[10] WU J J, HU W F, GONG Y, et al. Current pharmacological developments in 2, 3, 4', 5-tetrahydroxystilbene 2-O-β-D-glucoside (TSG)[J]. Eur J Pharmacol, 2017, 811:21-29.
[11] ZHANG W, CHEN X F, HUANG Y J, et al. 2, 3, 4', 5- Tetrahydroxystilbene-2-O-β-D-glucoside inhibits angiotensin Ⅱ-induced cardiac fibroblast proliferation via suppression of the reactive oxygen species-extracellular signal-regulated kinase 1/2 pathway[J]. Clin Exp Pharmacol Physiol, 2012, 39(5):429-437.
[12] YU F, XUE W, DONG L Y, et al. Tetrahydroxystilbene glucoside suppresses NAPDH oxidative stress to mitigate apoptosis and autophagy induced by cerebral ischemia/ reperfusion injury in mice[J]. Evid Based Complement Alternat Med, 2019, 2019:3913981.
[13] 雷娜, 张媛媛, 王齐. 二苯乙烯苷对脑缺血/再灌注损伤保护作用的研究进展[J]. 齐齐哈尔医学院学报, 2016, 37(32): 4075-4076.
[14] YU W H, ZHANG X D, WU H, et al. HO-1 is essential for tetrahydroxystilbene glucoside mediated mitochondrial biogenesis and anti-inflammation process in LPS-treated RAW264.7 macrophages[J]. Oxid Med Cell Longev, 2017, 2017:1818575.
[15] YAN B, XIE S B, LIU Z, et al. HDAC6 deacetylase activity is critical for lipopolysaccharide-induced activation of macrophages[J]. PLoS One, 2014, 9(10):e110718.
[16] LI Y H, WANG Y F, ZHU Y. Recent advances in anti-aging study of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucopyranoside:a main component of Polygonum multiflorum[J]. China J Chin Mater Med, 2016, 41(2):182-185.
[17] LIU Y, WANG Q, YANG J B, et al. Polygonum multiflorum thunb.:a review on chemical analysis, processing mechanism, quality evaluation, and hepatotoxicity[J]. Front Pharmacol, 2018, 9:364.
[18] 张雪. 何首乌二苯乙烯苷类成分研究进展[J]. 农业科技与装备, 2018(6):31-32.
[19] YU W H, ZHANG X D, LIU J Z, et al. Cyclosporine A suppressed glucose oxidase induced P53 mitochondrial translocation and hepatic cell apoptosis through blocking mitochondrial permeability transition[J]. Int J Biol Sci, 2016, 12(2):198-209.
[20] EL-KENAWI A, RUFFELL B. Inflammation, ROS, and mutagenesis[J]. Cancer Cell, 2017, 32(6):727-729.
[21] LEAVY O. Inflammation:regulating ROS[J]. Nat Rev Immunol, 2014, 14(6):357.
[22] MAEDA Y, INOGUCHI T. Oxidative stress and chronic inflammation[J]. Nippon Rinsho, 2016, 74(Suppl 2):73-76.
[23] SUZUKI T, YAMAMOTO M. Molecular basis of the Keap1- Nrf2 system[J]. Free Radic Biol Med, 2015, 88:93-100.
[24] CHEN D D, LI Z, BAO P Q, et al. Nrf2 deficiency aggravates Angiotensin Ⅱ-induced cardiac injury by increasing hypertrophy and enhancing IL-6/STAT3-dependent inflammation[J]. Biochim Biophys Acta Mol Basis Dis, 2019, 1865(6):1253-1264.
[25] LI S T, DAI Q, ZHANG S X, et al. Ulinastatin attenuates LPS-induced inflammation in mouse macrophage RAW264.7 cells by inhibiting the JNK/NF-κB signaling pathway and activating the PI3K/Akt/Nrf2 pathway[J]. Acta Pharmacol Sin, 2018, 39(8):1294-1304.
[26] QIAO Y Q, JIANG P F, GAO Y Z. Lutein prevents osteoarthritis through Nrf2 activation and downregulation of inflammation[J]. Arch Med Sci, 2018, 14(3):617-624.
[27] LEE I T, LUO S F, LEE C W, et al. Overexpression of HO-1 protects against TNF-alpha-mediated airway inflammation by down-regulation of TNFR1-dependent oxidative stress[J]. Am J Pathol, 2009, 175(2):519-532.
[28] XU X L, LI H Q, HOU X L, et al. Punicalagin induces Nrf2/ HO-1 expression via upregulation of PI3K/AKT pathway and inhibits LPS-induced oxidative stress in RAW264.7 macrophages[J]. Mediators Inflamm, 2015, 2015:380218.

二苯乙烯苷抗氧化和抗炎作用的机制研究

Antioxidant and anti-inflammatory effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in macrophages

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