&nbsp;Effects of resveratrol pretreatment on activation and inflammation of astrocytes after oxygen-glucose deprivation/reoxygenation injury in vitro

LIU Jing; LIU Jie; LIAO Hong-Yan; CHEN Yue; XIANG Qin; YANG Qin

doi:10.16098/j.issn.0529-1356.2020.03.001

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Acta Anatomica Sinica ›› 2020, Vol. 51 ›› Issue (3) : 313-319. DOI: 10.16098/j.issn.0529-1356.2020.03.001

Effects of resveratrol pretreatment on activation and inflammation of astrocytes after oxygen-glucose deprivation/reoxygenation injury in vitro

LIU Jing¹ LIU Jie¹LIAO Hong-yan¹ CHEN Yue¹ XIANG Qin^2* YANG Qin^1*

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Abstract

Objective To investigate the effects of resveratrol pretreatment on activation and inflammation of astrocytes after oxygen-glucose deprivation/reoxygenation (OGD/R) injury in vitro. Methods Twenty newborn SD rats within 24 hours were recruited,and their primary cultured astrocytes were deprived of oxygen glucose fpr 150 minutes and reoxygenation for 24 hours.The experment was divided into the normal (Nor),the control (Ctrl) and resveratrol pretreatment (Res) groups.Cell viability was detected with CCK-8 assay.Cell proliferation was measured with 5-ethynyl-2,-deoxyuridine (EdU).The expressions of glial fibrillary acidic protein (GFAP) and S100βproteins were determined with immunofluorescence.Theexpressions of GFAP,S100β,vimentin,interleukin (IL)-10,β-interferon (IFN-β),tumor necrosis factor α(TNF-α)and IL-1 β proteins were detected with Western blotting.The levels of IL-10,IFN-β,TNF-αand IL-1βproteins in the supernatant were measured by ELISA. Results CCK-8 assay showed that with the increase of resveratrol concentration (5, 20 and 40 μmol/L), the cell viability increased gradually, which was significantly lower than that of the control group (P<0.01). After that, with the increase of resveratrol concentration (80 and 100 μmol/L), the cell viability decreased significantly, and the cell viability of 100 μmol/L resveratrol group was significantly lower than in the control group (P<0.02). EdU testing showed that after OGD/R injury, the percentage of EdU positive cells in the control and resveratrol group was significantly higher than that of in the normal group (P<0.01), while it was significantly lower in the resveratrol pretreatment group than that of the control group (P<0.01).Immunofluorescence, Western blotting and ELISA method showed that the expressions or levels of GFAP, S100β, vimentin, IL-10, IFN-β, TNF-α, IL-1β proteins in the control and resveratrol group were significantly higher than that of the normal group (P<0.05), but the expresions or levels of IL-10 and IFN-β proteins in the resveratrol group were higher than those of the control group, and the others were lower than the control group (P<0.05).However，the expressions or levels of IL-10 and IFN-β proteins in the resveratrol group were higher than those of the control group, and the others in the resveratrol group were lower than those of the control group (P<0.05). Conclusion Resveratrol pretreatment can inhibit the activation of astrocytes and reduce the inflammation after OGD/R injury in vitro.

Key words

Resveratrol

/ Astrocyte / Activation / Inflammation / Western blotting / Enzyme-linked immunosobent assay / Immunofluorescence / Rat

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LIU Jing LIU Jie LIAO Hong-yan CHEN Yue XIANG Qin YANG Qin. Effects of resveratrol pretreatment on activation and inflammation of astrocytes after oxygen-glucose deprivation/reoxygenation injury in vitro[J]. Acta Anatomica Sinica. 2020, 51(3): 313-319 https://doi.org/10.16098/j.issn.0529-1356.2020.03.001

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