Role of NF-κB in cytochrome P450 epoxygenases down-regulation during an inflammatory process in astrocytes

• Published in: Neurochemistry international 2019-10, Vol.129, p.104499-104499, Article 104499
• Main Authors: Navarro-Mabarak, Cynthia, Mitre-Aguilar, Irma Beatriz, Camacho-Carranza, Rafael, Arias, Clorinda, Zentella-Dehesa, Alejandro, Espinosa-Aguirre, Jesús Javier
• Format: Article
• Language: English
• Subjects: Animals; Aryl Hydrocarbon Hydroxylases; Astrocytes; Astrocytes - metabolism; Benzamides - pharmacology; Cells, Cultured; Cerebral Cortex - cytology; Cytochrome P-450 Enzyme System; Cytochrome P450; Cytochrome P450 Family 2; Down-Regulation - drug effects; Eicosanoids - biosynthesis; Endotoxins - pharmacology; Epoxygenase; Gene Expression Regulation; Inflammation; Inflammation - chemically induced; Inflammation - genetics; Inflammation - metabolism; Male; NF-kappa B - antagonists & inhibitors; NF-kappa B - physiology; NF-κB; Primary Cell Culture; Promoter Regions, Genetic; Rats; Rats, Wistar; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Steroid 16-alpha-Hydroxylase; Tumor Necrosis Factor-alpha - biosynthesis; Tumor Necrosis Factor-alpha - genetics
• ISSN: 0197-0186; 1872-9754
• DOI: 10.1016/j.neuint.2019.104499

Cytochrome P450 (CYP) epoxygenases and their metabolic products, epoxyeicosatrienoic acids (EETs), have been proposed as important therapeutic targets in the brain. However, CYP expression can be modified by the presence of diverse pro-inflammatory cytokines and the subsequent activation of the NF-κB pathway. It has been indicated that CYP epoxygenases are down-regulated by inflammation in the heart, kidney and liver. However, up to this point, there has been no evidence regarding regulation of CYP epoxygenases during inflammation in the brain. Therefore, in order to explore the effects of inflammation and NF-κB activation in CYP2J3 and CYP2C11 regulation, rat primary astrocytes cultures were treated with LPS with and without IMD-0354 (selective NF-κB inhibitor). Cyp2j3 and Cyp2c11 mRNA expression was determined by qRT-PCR; protein expression was determined by immunofluorescence and by Western Blot and total epoxygenase activity was determined by the quantification of EETs by ELISA. NF-κB binding sites in Cyp2j3 and Cyp2c11 promoter regions were bioinformatically predicted and Electrophoretic Mobility Shift Assays (EMSA) were performed to determine if each hypothetic response element was able to bind NF-κB complexes. Results shown that LPS treatment is able to down-regulate astrocyte CYP2J3 and CYP2C11 mRNA, protein and activity. Additionally, we have identified NK-κB as the transcription factor involved in this regulation. [Display omitted] •Inflammation in astrocytes could down-regulate CYP2J3 and CYP2C11 mRNA, protein and activity.•Inhibition of NF-κB prevented the down-regulation of CYP2J3 mRNA and protein.•Inhibition of NF-κB partially prevented the down-regulation of CYP2C11 mRNA and protein.•Inhibition of NF-κB completely prevented the down-regulation of EETs production.•NF-κB is able to bind to at least two response elements in the promoter of Cyp2j3 and Cyp2c11.