Increased PUFA levels in kidney epithelial cells in the course of diclofenac toxicity. (August 2020)

• Record Type: Journal Article
• Authors: Aslan, Mutay; Kırımlıoğlu, Esma; Afşar, Ebru; Çeker, Tuğçe; Yılmaz, Çağatay
• Is Part Of: Toxicology in vitro. Volume 66(2020)
• Publication Date: 2020-08
• Subjects: Diclofenac -- Kidney cells -- Thymoquinone -- Polyunsaturated fatty acids; AA arachidonic acid -- COX cyclooxygenase -- cPLA2 cytosolic phospholipase A2 -- DCL diclofenac -- DGLA dihomo-gamma-linolenic acid -- DHA docosahexaenoic acid -- LOX lipoxygenase -- DMEM Dulbecco's Modified Eagle's Medium -- DTNB 5, 5′-dithiobis-2-nitrobenzoic acid -- EPA eicosapentaenoic acid -- MRM multiple reaction monitoring -- MS/MS tandem mass spectrometry -- MTT 3-(4, 5dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide -- NSAID nonsteroidal anti-inflammatory drug -- PGE2 prostaglandin E2 -- PUFAs polyunsaturated fatty acids -- TQ thymoquinone -- UFLC ultrafast liquid chromatography; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2020.104836
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: This study evaluated polyunsaturated fatty acids (PUFAs) in human kidney epithelial cells exposed to diclofenac (DCL) toxicity. Kidney cells were treated with DCL to induce cytotoxicity and thymoquinone (TQ) was administered to decrease cytotoxic effects. Levels of arachidonic acid (AA, C20:4n-6), dihomo-gamma-linolenic acid (DGLA, C20:3n-6), eicosapentaenoic acid (EPA, C20:5n-3) and docosahexaenoic acid (DHA, C22:6n-3) were determined by liquid chromatography coupled with tandem mass spectrometry. Cytosolic phospholipase A2 (cPLA2 ), cyclooxygenase 1 (COX-1) and prostaglandin E2 (PGE2 ) were measured to evaluate changes in enzyme activity. Immunofluorescence staining and western blot analysis was performed to determine protein levels of COX- 1. Renal cell toxicity was accomplished by DCL and was alleviated by TQ treatment. Diclofenac significantly increased all measured PUFAs while pretreatment with TQ decreased PUFA levels in DCL treated cells. Cytosolic PLA2 and total COX activity was significantly decreased in DCL treated cells. Immunofluorescence staining and western blot analysis confirmed significantly decreased COX-1 levels in DCL and DCL + TQ treated groups. The results of this study reveal that DCL treatment is associated with accumulation of PUFAs in kidney cells. We suggest that PUFA accumulation in DCL toxicity might be a consequence of both cPLA2 and COX-1 inhibition. Thymoquinone administration, along with DCL treatment alleviated the buildup of PUFAs and DCL-induced cell death in kidney cells. Highlights: Diclofenac (DCL) toxicity causes polyunsaturated fatty acid (PUFA) accumulation in kidney cells. PUFA accumulation in DCL toxicity is associated with cytosolic phospholipase A2 inhibition PUFA accumulation in DCL toxicity is associated with cyclooxygenase 1 inhibition Thymoquinone attenuates diclofenac-induced PUFA accumulation.