Arachidonic acid alleviates the detrimental effects of acetylsalicylic acid on human granulosa cells performance in vitro

Masoumeh Khajeh, Mohammad Nouri, Aalie Ghasemzadeh, Amir Mehdizadeh, Dariush Shanehbandi, Soudabe Yousefi, Masoud Darabi*, Reza Rahbarghazi

*Corresponding author for this work
3 Citations (Scopus)

Abstract

Here, we investigated the biological effects of arachidonic acid (AA) in human cumulus granulosa cells (CGCs) after exposure to ASA. Cells were isolated from the follicular fluid and incubated with 0.5 mM acetylsalicylic acid (ASA) and 50 µM AA. Cell viability was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. E2 and P4 levels were measured by chemiluminescence assay. Expression of genes including CYP19A1, FACN, and SCD1 was measured by real-time polymerase chain reaction assay. Oxidative status was analyzed by monitoring glutathione peroxidase activity. The fatty acid profile was analyzed by the gas chromatography technique. Enzyme-linked immunosorbent assay was used to measure prostaglandin E2 (PGE2) in CGCs after exposure to ASA and AA. Protein levels of the estrogen receptor were studied by immunofluorescence staining. Ultrastructural changes were evaluated by transmission electron microscopy imaging. ASA treatment reduced E2 production, Cyp19a1 expression, glutathione peroxidase (GPx) activity, and estradiol receptor expression in CGCs. The addition of AA prevented the ASA-induced E2 reduction (p <.05) and expression of Cyp19a1. Moreover, AA increased the antioxidant capacity of CGCs exposed to ASA by promoting GPx activity (p <.05). AA increased monounsaturated fatty acid/saturated fatty acid ratio compared with the ASA group (p <.05). AA supplementation triggered the synthesis and secretion of PGE2 in ASA-treated CGCS (p <.05). Cytoplasmic vacuolation observed in the ASA group and treatment with AA intensified vacuolation rate. The expression of the estrogen receptor was increased after AA supplementation. Data demonstrated that AA decreased the detrimental effects of ASA on human CGCs after 72 hr.

Original languageEnglish
JournalMolecular Reproduction and Development
Volume87
Issue number5
Pages (from-to)607-619
Number of pages13
ISSN1040-452X
DOIs
Publication statusPublished - 01.05.2020

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