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

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。

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

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.