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Omp31 of Brucella Inhibits NF-κB p65 Signaling Pathway by Inducing Autophagy in BV-2 Microglia

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Abstract

Neurobrucellosis is a serious central nervous system (CNS) inflammatory disorder caused by Brucella, and outer membrane protein-31 (Omp31) plays an important role in Brucella infection. This study aims to determine whether Omp31 can induce autophagy in BV-2 microglia. Another goal of the study is to further examine the effect of autophagy on the nuclear transcription factor κB (NF-κB) p65 signaling pathway. We observed that Omp31 stimulated autophagy by increasing microtubule-associated protein 1 light chain 3B (LC3B-II) levels and inducing autophagosome formation at 6 h and 12 h. Concomitantly, Omp31 induced tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) expression in a time-dependent manner but reduced the expression of TNF-α at 6 h. We utilized Omp31 with or without rapamycin or 3-methyladenine (3-MA) to treat BV-2 microglia, and it demonstrated further that Omp31 induced autophagy by promoting LC3B-II, Beclin-1 proteins expression and inhibiting the p62 protein levels. Furthermore, we explored the effects of autophagy on the NF-κB p65 pathway through western blot analysis, RT-qPCR assay, enzyme-linked immunosorbent assay (ELISA) and immunofluorescence. The data suggest that Omp31 as well as rapamycin, the autophagy inducer, can decrease TNF-α levels through the inhibition of the NF-κB p65 signaling pathway. Taken together, Omp31 can function as a catalyst in both autophagy induction and NF-κB p65 signal inhibition. Furthermore, Omp31-induced autophagy may inhibit the expression of TNF-α by negatively regulating NF-κB p65 signaling pathway.

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Funding

This study was funded by the National Natural Science Foundation of China (Grant No. 31660030), the Key Research and Development Project of Ningxia (Grant No. 2018BFG02017), Key scientific research projects of Ningxia Health and Family Planning Commission (Grant No. 2018-NW-009).

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Author notes

  1. Zhao Wang and Guowei Wang have contributed equally to this work.

Authors and Affiliations

  1. Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, China

    Zhao Wang

  2. Neurology Center, The General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, China

    Guowei Wang, Yanbai Wang, Qiang Liu, Haining Li & Zhenhai Wang

  3. The First Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Peng Xie

  4. Diagnosis and Treatment Engineering Technology Research Center of Nervous System Diseases of Ningxia Hui Autonomous Region, Yinchuan, Ningxia Hui Autonomous Region, China

    Zhenhai Wang

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  1. Zhao Wang
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Wang, Z., Wang, G., Wang, Y. et al. Omp31 of Brucella Inhibits NF-κB p65 Signaling Pathway by Inducing Autophagy in BV-2 Microglia. Neurochem Res 46, 3264–3272 (2021). https://doi.org/10.1007/s11064-021-03429-4

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