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白藜芦醇苷通过上调SIRT1抑制氧化低密度脂蛋白诱导的THP-1巨噬细胞增殖和炎性因子表达 被引量:2

Polydatin inhibits cell proliferation and expressions of inflammatory cytokines in THP-1 cells induced by ox-LDL via up-regulating SIRTI
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摘要 目的 观察白藜芦醇苷对氧化低密度脂蛋白(ox-LDL)诱导THP-1巨噬细胞(MDM)炎性增殖与细胞因子表达的影响及机制。方法 培养THP-1巨噬细胞,分为对照组(仅加入普通培养基)、ox-LDL组(80μmol/L ox-LDL处理细胞24h)、白藜芦醇苷处理组(100 μmol/L 白藜芦醇苷预处理2 h后,再加入80μmol/L ox-LDL处理24 h)、EX-527组(10 μmol/L EX-527预处理细胞2h)。CCK-8法检测各组细胞增殖率, 分光光度计检测细胞内超氧化物歧化酶(SOD)以及丙二醛(MDA)含量, 2′, 7′-二氯二氢荧光素二乙酸酯(DCFH-DA)负载法检测活性氧(ROS)的含量; 实时定量PCR法检测白藜芦醇苷对组蛋白去乙酰化酶沉默信息调节因子1(SIRT1)、单核细胞趋化因子1(MCP-1)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6) mRNA水平的影响,Western blot法检测SIRT1、MCP-1、TNF-α、IL-6的蛋白水平。结果 ox-LDL促进THP-1巨噬细胞增殖。100μmol/L白藜芦醇苷组明显抑制ox-LDL诱导的细胞增殖, 同时明显增加SOD含量, 降低胞内MDA与ROS含量。ox-LDL抑制THP-1巨噬细胞中SIRT1 mRNA与蛋白表达, 促进MCP-1、TNF-α、IL-6mRNA与蛋白表达。白藜芦醇苷显著增加ox-LDL诱导后SIRT1 mRNA与蛋白表达, 抑制MCP-1、TNF-α、IL-6 mRNA与蛋白表达, EX-527能抑制白藜芦醇苷上述保护作用。结论白藜芦醇苷可通过上调SIRT1表达提高抗巨噬细胞增殖的能力, 减少ox-LDL诱导细胞内活性氧生成、抑制炎性因子表达。 Objective To investigate the effects of polydatin on cell proliferation and cytokine expression in THP-1 monocyte-derived macrophages (MDMs) induced by oxidized low-density lipoprotein (ox-LDL), and the possible mechanisms. Methods MDMs were divided into the control group ( only treated with ordinary culture medium), ox-LDL group (treated wtih 80 μmol/L ox-LDL for 24 hours), polydatin treatment group (treated with 100 μmol/L polydatin for 2 hours prior to the treatment with 80μmol/LL ox-LDL for 24 hours) and EX-527 treatment group (treated with 10 μmol/L SIRT1 inhibitor EX-527 for 2 hours prior to the treatment with ox-LDL and polydatin ). The effects of polydatin on ox-LDL-induced oxidative proliferation and cytokine expression in MDMs were evaluated by CCK-8 assay. Spectrofluorometry was used to determine the intracellular level of superoxide dismutase (SOD) and malondialdehyde (MDA): DCFH-DA loading was used to detect the content of reactive oxidative species (ROS). The levels of silent mating type information regulator 1 ( SIRT1 ), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were examined by real-time quantitative PCR and Western blotting. Results Polydatin (100 μmol/L) inhibited the proliferation of MDMs induced by ox-LDL, decreased the levels of MDA and ROS, whereas the level of SOD increased. The mRNA and protein levels of SIRT1 in MDMs were inhibited by ox-LDL, whereas the expressions of MCP-1, TNF-α and IL-6 were promoted. Pre-treatment with EX-527 attenuated the inhibitory effects of polydatin on the proliferation of MDMs, inhibited the expressions of SIRT1, promoted the expressions of MCP-1, TNF-α and IL-6. Conclusion Polydatin up-regulates the expression of SIRT1 to increase the ability of anti-macrophage proliferation, reduce the level of the intracellular ROS induced by ox-LDL, and inhibit the expression of inflammatory cytokines.
作者 马懿 阮云军 王玉筵 吴赛珠 MA Yi1, RUAN Yunjun2, WANG Yuyan3, WU Saizhu3(1Department of Cardiology, Affiliated Hospital, Zunyi Medical University, Zunyi 563000; 2Department of Cardiology, Guangzhou Military General Hospital, Guangzhou 510010; 3Deartment of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China)
出处 《细胞与分子免疫学杂志》 CSCD 北大核心 2018年第3期193-198,共6页 Journal of Cellular and Molecular Immunology
基金 国家“973”项目子课题(2007CB507404) 贵州省科技计划项目(黔科合LH字[2017]7109号) 贵州省中医药管理局科学技术研究项目(QZYY2017-096) 遵医附院博士科研基金(院字2015-02)
关键词 白藜芦醇苷 氧化低密度脂蛋白(ox—LDL) 单核细胞趋化因子1(MCP-1) 活性氧(ROS) 沉默信息调节因子1(SIRT1) polydatin oxidized low-density lipoprotein (ox-LDL) monocyte chemoattractant protein-1 MCP-1) reactiveoxygen species (ROS) silent mating type information regulator 1 (SIRT1)
作者简介 马懿(1980-),男,贵州遵义人,副主任医师,博士Tel:13638528953;E—mail:mayimayizmc@126.com。通讯作者,马懿
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