白藜芦醇对氧糖剥夺/再复氧损伤后体外小胶质细胞极化的影响

doi:10.16098/j.issn.0529-1356.2020.03.002

解剖学报 ›› 2020, Vol. 51 ›› Issue (3): 320-325.doi: 10.16098/j.issn.0529-1356.2020.03.002

白藜芦醇对氧糖剥夺/再复氧损伤后体外小胶质细胞极化的影响

刘杰¹任瑜¹ 陈月¹ 刘菁¹ 向勤^2*杨琴^1*

1. 重庆医科大学附属第一医院神经内科，重庆 400016； 2. 重庆市长寿区人民医院神经内科，重庆 401220

收稿日期:2019-11-08 修回日期:2020-02-11 出版日期:2020-06-06 发布日期:2020-06-06
通讯作者: 向勤;杨琴 E-mail:xyqh200@126.com
基金资助:
国家自然科学基金面上项目;西藏自治区自然科学基金项目

Effects of resveratrol on microglia polarization after oxygen glucose deprivation/reoxygenation injury in vitro

LIU Jie¹ REN Yu¹ CHEN Yue¹ LIU Jing¹ XIANG Qin^2* YANG Qin^1*

1.Department of Neurology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China;2.Department of Neurology, Longevity District People's Hospital of Chongqing, Chongqing 401220, China

Received:2019-11-08 Revised:2020-02-11 Online:2020-06-06 Published:2020-06-06
Contact: XIANG Qin;YANG Qin E-mail:xyqh200@126.com
Supported by:
the National Natural Science Foundation of China

摘要/Abstract

摘要：

目的探讨白藜芦醇对氧糖剥夺/再复氧损伤（OGD/R）后小胶质细胞极化的影响。方法对体外培养的N9小胶质细胞进行氧糖剥夺150 min，复氧培养24 h。实验分为正常组、对照组和白藜芦醇预处理24 h组。细胞计数盒-8(CCK-8)法检测细胞活力，硫代巴比妥酸（TBA）和水溶性四唑盐（WST-1）法分别检测细胞上清液中丙二醛（MDA）含量和总超氧化物歧化酶（SOD）活性，免疫荧光法检测核因子-E2-相关因子2（Nrf2）的核移位，Western blotting和Real-time PCR法检测CD206、诱导型一氧化氮合酶（iNOS）、Nrf2、血红素加氧酶1（HO-1）和苯醌还原酶（NQO1）蛋白和mRNA表达水平。结果 OGD/R损伤后，白藜芦醇组细胞活力、SOD活力、CD206、Nrf2、HO-1、NQO1蛋白或mRNA表达均显著高于对照组（P<0.05,n=3），而MDA含量和iNOS蛋白或mRNA表达均显著低于对照组（P<0.05,n=3），白藜芦醇组Nrf2 蛋白较对照组明显移位到细胞核。结论白藜芦醇预处理可能通过增强Nrf2的激活，调控M1/M2型小胶质细胞极化，从而减轻OGD/R后小胶质细胞的氧化应激损伤。

关键词: 白藜芦醇, 小胶质细胞, 极化, 核因子-E2-相关因子2, 氧化应激, 免疫印迹法, 实时定量聚合酶链反应

Abstract:

Objective To investigate the effect of resveratrol on microglia polarization after oxygen glucose deprivation/reoxygenation (OGD/R) in vitro. Methods N9 microglias were cultured under oxygen-glucose deprivation for 150 minutes and reoxygenation for 24 hours.The experiment was divided into normal, control and resveratrol pretreatment groups. Cell viability was measured by cell counting kit-8(CCK-8) assay. Malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity in the supernatant were detected by thiobarbituric acid(TBA) and water soluble terazolium salt(WST-1) respectively. Nuclear factor E2-related factor 2 (Nrf2) nuclear translocation was detected by immunofluorescence. The levels of CD206, inducible nitric oxide synthase (iNOS), Nrf2, heme oxygenase-1(HO-1), NAD(P)H: quinone oxidoreductase 1(NQO1) protein were measured by Western blotting. The mRNA levels of CD206, iNOS were detected by Real-time PCR. Results After OGD/R injury, cell viability, SOD activity, CD206, Nrf2, HO-1, NQO1 protein or mRNA expression in resveratrol groups were significantly higher than those in the control group (P<0.05, n=3), while MDA content and iNOS protein and mRNA expression were significantly lower than those in the control group (P<0.05, n=3). Nrf2 protein in resveratrol group was significantly transferred to the nucleus compared with the control group. Conclusion Resveratrol pretreatment may regulate M1/M2 polarization of microglia and reduce oxidative stress after OGD/R via strengthening activation of Nrf2.

Key words: Resveratrol, Microglia, Polarization, Nuclear factor E2-related factor 2, Oxidative stress, Western blotting, Real-time PCR

中图分类号:

刘杰任瑜陈月刘菁向勤杨琴. 白藜芦醇对氧糖剥夺/再复氧损伤后体外小胶质细胞极化的影响[J]. 解剖学报, 2020, 51(3): 320-325.

LIU Jie REN Yu CHEN Yue LIU Jing XIANG Qin YANG Qin. Effects of resveratrol on microglia polarization after oxygen glucose deprivation/reoxygenation injury in vitro[J]. Acta Anatomica Sinica, 2020, 51(3): 320-325.

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白藜芦醇对氧糖剥夺/再复氧损伤后体外小胶质细胞极化的影响

Effects of resveratrol on microglia polarization after oxygen glucose deprivation/reoxygenation injury in vitro

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