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; 任瑜; 陈月; 刘菁; 向勤; 杨琴

doi:10.16098/j.issn.0529-1356.2020.03.002

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

神经生物学

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*

作者信息 +

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*

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2020.03.002

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 https://doi.org/10.16098/j.issn.0529-1356.2020.03.002

中图分类号： R322.81 R743.3

参考文献

［1］ Sharma A, Liaw K, Sharma R, et al. Targeting mitochondrial dysfunction and oxidative stress in activated microglia using dendrimer-based therapeutics［J］. Theranostics, 2018, 8 (20):5529-5547.

［2］ Kanazawa M, Ninomiya Ⅰ, Hatakeyama M, et al. Microglia and monocytes/macrophages polarization reveal novel therapeutic mechanism against stroke ［J］. Int J Mol Sci, 2017,18(10): E2135.

［3］ Yang X, Xu S, Qian Y, et al. Resveratrol regulates microglia M1/M2 polarization via PGC-1α in conditions of neuroinflammatory injury ［J］. Brain Behav Immun, 2017, 64:162-172.

［4］ Lopez MS, Dempsey RJ, Vemuganti R. Resveratrol neuroprotection in stroke and traumatic CNS injury ［J］. Neurochem Int, 2015, 89:75-82.

［5］ Liao HY, Liu J, Liu J, et al. Effect of resveratrol on activation of microglia cell line N9 after oxygen-glucose deprivation/reoxygenation injury in vitro ［J］. Acta Anatomica Sinica, 2019,50(2): 137-143. (in Chinese)

廖鸿雁，刘杰，刘菁，等. 白藜芦醇对氧糖剥夺/再复氧损伤后小胶质细胞系N9 活化的影响［J］. 解剖学报，2019； 50(2)：137-143.

［6］ Ma S, Fan L, Li J, et al. Resveratrol promoted the M2 polarization of microglia and reduced neuroinflammation after cerebral ischemia by inhibiting miR-155［J］. Int J Neurosci, 2019, 20:1-13.

［7］ Zhang R, Xu M, Wang Y, et al. Nrf2-a promising therapeutic target for defensing against oxidative stress in stroke ［J］. Mol Neurobiol, 2017,54(8):6006-6017.

［8］ Ren J, Fan C, Chen N, et al. Resveratrol pretreatment attenuates cerebral ischemic injury by upregulating expression of transcription factor Nrf2 and HO-1 in rats ［J］. Neurochem Res, 2011, 36(12):2352-2362.

［9］ Shen C, Cheng W, Yu P, et al. Resveratrol pretreatment attenuates injury and promotes proliferation of neural stem cells after oxygen-glucose deprivation/reoxygenation by upregulating expressions of Nrf2, HO-1 and NQO1 in vitro ［J］. Mol Med Rep, 2016, 14(4): 3646-3654.

［10］ Wang N, Liang H, Zen K, et al. Molecular mechanisms that influence the macrophage m1-m2 polarization balance ［J］. Front Immunol, 2014, 5:614.

［11］ Hu X, Li P, Guo Y, et al. Microglia/macrophage polarization dynamics reveal novel mechanism of injury expansion after focal cerebral ischemia ［J］. Stroke, 2012, 43（11）: 3063-3070.

［12］ Yu PP，Wang L，Tang FR，et al.Effect of resveratrol on activation of astrocyte after cerebral ischemia/reperfusion injury in rats ［J］.Chinese Pharmacological Bulletin, 2015，31 (9): 1228-1233.(in Chinese)

余萍萍，王莉，唐凡人，等. 白藜芦醇对大鼠脑缺血/再灌注损伤后星形胶质细胞活化的影响［J］.中国药理学通报，2015，31(9): 1228-1233.

［13］ Waza AA, Hamid Z, Ali S, et al. A review on heme oxygenase-1 induction: is it a necessary evil ［J］? Inflamm Res, 2018, 67(7):579-588.

［14］ Liang M, Wang Z, Li H, et al. l-Arginine induces antioxidant response to prevent oxidative stress via stimulation of glutathione synthesis and activation of Nrf2 pathway ［J］. Food Chem Toxicol, 2018, 115:315-328.

［15］ Kim EN, Lim JH, Kim MY, et al. Resveratrol, a Nrf2 activator, ameliorates aging-related progressive renal injury ［J］. Aging (Albany NY), 2018,10(1):83-99.

［16］ Wang Y, Huang Y, Xu Y, et al. A dual AMPK/Nrf2 activator reduces brain inflammation after stroke by enhancing microglia M2 polarization ［J］. Antioxid Redox Signal, 2018,28(2):141-163.

［17］ Jin MM, Wang F, Qi D, et al. A Critical role of autophagy in regulaing microglia polarization in neurodegeneration［J］.Front Aging Neurosci, 2018,10:378.