NMDA receptor inhibitor MK801 alleviated pro-inflammatory polarization of BV-2 microglia cells

Chih-Cheng Wu; Chung-Yuh Tzeng; Cheng-Yi Chang; Jiaan-Der Wang; Yu-Fang Chen; Wen-Ying Chen; Yu-Hsiang Kuan; Su-Lan Liao; Wen-Yi Wang; Chun-Jung Chen

doi:10.1016/j.ejphar.2023.175927

NMDA receptor inhibitor MK801 alleviated pro-inflammatory polarization of BV-2 microglia cells

Eur J Pharmacol. 2023 Sep 15:955:175927. doi: 10.1016/j.ejphar.2023.175927. Epub 2023 Jul 20.

Authors

Chih-Cheng Wu¹, Chung-Yuh Tzeng², Cheng-Yi Chang³, Jiaan-Der Wang⁴, Yu-Fang Chen⁵, Wen-Ying Chen⁶, Yu-Hsiang Kuan⁷, Su-Lan Liao⁸, Wen-Yi Wang⁹, Chun-Jung Chen¹⁰

Affiliations

¹ Department of Anesthesiology, Taichung Veterans General Hospital, Taichung City, 407, Taiwan; Department of Financial Engineering, Providence University, Taichung City, 433, Taiwan; Department of Data Science and Big Data Analytics, Providence University, Taichung City, 433, Taiwan.
² Department of Orthopedics, Taichung Veterans General Hospital, Taichung City, 407, Taiwan; Department of Medicinal Botanicals and Health Applications, Da-Yeh University, Changhua, 515, Taiwan.
³ Department of Surgery, Feng Yuan Hospital, Taichung City, 420, Taiwan; Department of Veterinary Medicine, National Chung Hsing University, Taichung City, 402, Taiwan.
⁴ Children's Medical Center, Taichung Veterans General Hospital, Taichung City, 407, Taiwan; Department of Industrial Engineering and Enterprise Information, Tunghai University, Taichung City, 407, Taiwan.
⁵ Department of Microbiology & Immunology, National Cheng Kung University, Tainan City, 701, Taiwan.
⁶ Department of Veterinary Medicine, National Chung Hsing University, Taichung City, 402, Taiwan.
⁷ Department of Pharmacology, Chung Shan Medical University, Taichung City, 402, Taiwan.
⁸ Department of Medical Research, Taichung Veterans General Hospital, Taichung City, 407, Taiwan.
⁹ Department of Nursing, HungKuang University, Taichung City, 433, Taiwan.
¹⁰ Department of Medical Research, Taichung Veterans General Hospital, Taichung City, 407, Taiwan; Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung City, 404, Taiwan. Electronic address: cjchen@vghtc.gov.tw.

PMID: 37479018
DOI: 10.1016/j.ejphar.2023.175927

Abstract

Microglia have both protective and pathogenic properties, while polarization plays a decisive role in their functional diversity. Apart from being an energetic organelle, mitochondria possess biological capabilities of signaling and immunity involving mitochondrial dynamics. The N-methyl-D-aspartate (NMDA)-type glutamate receptor displays excitatory neurotransmission, excitatory neurotoxicity and pro-inflammatory properties in a membrane location- and cell context-dependent manner. In this study, we have provided experimental evidence showing that by acting on mitochondrial dynamics, NMDA receptors displayed pro-inflammatory properties, while its non-competitive inhibitor MK801 exhibited anti-inflammatory potential in Lipopolysaccharide (LPS)-challenged BV-2 microglia cells. LPS stimulation increased the protein phosphorylation of cells regarding their NMDA receptor component subunits and Calcium/Calmodulin-dependent Protein Kinase II (CaMKII), along with mobilizing intracellular calcium. Additionally, parallel changes occurred in the activation of Transforming Growth Factor-β (TGF-β)-Activated Kinase 1 (TAK1), NF-κB p65 and NF-κB DNA binding activity, acquisition of pro-inflammatory M1 polarization and expression of pro-inflammatory cytokines. LPS-treated cells further displayed signs of mitochondrial dysfunction with higher expressions of the active form of Dynamin-Related Protein 1 (Drp1), NADPH Oxidase-2 (NOX2) expression and the generation of DCFDA-/MitoSOX-sensitive Reactive Oxygen Species (ROS). NMDA receptor blockade by MK801, along with CaMKII inhibitor KN93, Drp1 inhibitor Mdivi-1 and antioxidant apocynin alleviated LPS-induced pro-inflammatory changes. Other than the reported CaMKII/TAK1/NF-κB axis, our in vitro study revealed the CaMKII/Drp1/ROS/NF-κB axis being an alternative cascade for shaping pro-inflammatory phenotypes of microglia upon LPS stimulation, and MK801 having the potential for inhibiting microglia activation and any associated inflammatory damages.

Keywords: Inflammation; Mitochondrial dynamics; NMDA receptor; Polarization.