Nrf2通路对日本脑炎病毒感染小鼠神经瘤细胞引起氧化应激反应的影响

doi:10.11843/j.issn.0366-6964.2023.01.029

Abstract

Abstract: To investigate the role of the Nrf2/HO-1 pathway in neuronal damage caused by Japanese encephalitis virus (JEV) infection, an in vitro infection model was developed in which Nrf2 agonist tert-butylhydroquinone (TBHQ) was used to treat JEV-infected mouse neuroblastoma (N2a) cells. N2a cells were divided into following four groups: DMEM control group (Control), JEV-infected group (JEV), TBHQ-treated group (TBHQ), and JEV-infected +TBHQ -treated group (JEV+TBHQ). The cytopathic lesions were observed after different treatments and the changes in ROS levels and MDA, SOD and CAT contents were detected. The mRNA and protein expression of Nrf2 and HO-1 were detected by qPCR and WB techniques, and Nrf2 protein expression in each group of cells was detected by immunofluorescence, and inflammatory factors and viral content were measured. The results showed that the level of ROS gradually increased with time after JEV infection of N2a cells and was reached the highest at 36 h. The mRNA of Nrf2 and HO-1 increased at 36 h after JEV infection, and the level of inflammatory factors increased. JEV infection of N2a cells resulted in cell crumpling, cell membrane rupture, increased ROS and MDA levels and decreased SOD and CAT levels; treatment with 40 μmol·L^-1 TBHQ for 6 h reduced cell damage, decreased ROS and MDA levels and increased SOD and CAT activities. Both JEV and TBHQ activated the mRNA and protein expression of Nrf2 and HO-1, and the expression levels of proteins related to the Nrf2/HO-1 pathway were significantly increased by combined treatment with JEV+TBHQ, and the nucleation of Nrf2 in the cells by immunofluorescence was evident, and the mRNA levels of TNF-α and IL-6 and the levels of viruses decreased. The above results suggest that TBHQ is able to reduce cellular oxidative stress and inflammation levels induced by JEV through the Nrf2/HO-1 pathway.

Key words: Japanese encephalitis virus, oxidative stress, TBHQ, N2a cells, Nrf2/HO-1 signalling pathway

CLC Number:

S852.3

LIU Zelin, GUO Xiaoyan, LI Jiahuan, GAO Mingxing, CHENG Guofu, HU Xueying, ZHANG Wanpo, GU Changqin. Effect of the Nrf2 Pathway on Oxidative Stress Induced by Japanese Encephalitis Virus Infection of Mouse Neuroblastoma Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 317-327.

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Effect of the Nrf2 Pathway on Oxidative Stress Induced by Japanese Encephalitis Virus Infection of Mouse Neuroblastoma Cells

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