Effect of resveratrol on activation of microglia cell line N9 after oxygen-glucose deprivation/reoxygenation injury in vitro

LIAO Hong-yan LIU Jie LIU Jing GUO Shuang YANG Qin*

doi:10.16098/j.issn.0529-1356.2019.02.001

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Acta Anatomica Sinica ›› 2019, Vol. 50 ›› Issue (2) : 137-144. DOI: 10.16098/j.issn.0529-1356.2019.02.001

Effect of resveratrol on activation of microglia cell line N9 after oxygen-glucose deprivation/reoxygenation injury in vitro

LIAO Hong-yan LIU Jie LIU Jing GUO Shuang YANG Qin*

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Abstract

Objective To investigate the effect of resveratrol on the activation of microglia cell line N9 after oxygen-glucose deprivation/reoxygenation (OGD/R) injury in vitro.Methods N9 microglias were cultured under oxygen-glucose deprivation for 150 minutes and reoxygenation for 24 hours. The experiments were divided into normal (Nor), control (Ctrl) and resveratrol pretreatment group (Res). Cell viability was determined by cell counting kit-8(CCK-8) assay. The percentage of apoptosis was measured by flow cytometry. Expression of ionized calcium binding adaptor molecule-1(Iba1), CD11b, Caspase-3, Bax，interleukin-10 (IL-10), tumor necrosis factor α(TNF-α) and IL-1β proteins were detected by immunofluorescence. The levels of IL-10, TNF-α and IL-1βprotein in the supernatant were measured by ELISA. Results CCK-8 assay showed that the viability of microglia in the control and 1,5,20,40,80 μmol/L resveratrol groups significantly decreased compared with the normal group after OGD/R injury (P＜0.05，n=3), and significantly enhanced in the 5, 20, and 40 μmol/L resveratrol group than those in the control group, the peak was 20 μmol/L resveratrol group (P＜0.05,n=3). Immunofluorescence, Western blotting and ELISA or flow cytometry method showed that expressions or levels of CD11b, Iba1, Caspase3, Bax, IL-10, TNF-α, IL-1β proteins and the percent of apoptosis significantly increased in the control and the resveratrol group after OGD/R injury (P<0.05, n=3). However, except the expression or level of IL-10 protein was higher, the rests were lower in the resveratrol group than those of the control group (P<0.05, n=3). Conclusion Resveratrol pretreatment can inhibit the activation and apoptosis of microglia, reduce inflammation after OGD/R.

Key words

Resveratrol / Microglia / Activation / Inflammation / Western blotting

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LIAO Hong-yan LIU Jie LIU Jing GUO Shuang YANG Qin. 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): 137-144 https://doi.org/10.16098/j.issn.0529-1356.2019.02.001

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