Arachidonic acid alleviates the detrimental effects of acetylsalicylic acid on human granulosa cells performance in vitro.,Molecular Reproduction and Development

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Arachidonic acid alleviates the detrimental effects of acetylsalicylic acid on human granulosa cells performance in vitro.
Molecular Reproduction and Development ( IF 2.7 ) Pub Date : 2020-04-08 , DOI: 10.1002/mrd.23343
Masoumeh Khajeh _{1,

2} , Mohammad Nouri ₁ , Aalie Ghasemzadeh ₃ , Amir Mehdizadeh ₄ , Dariush Shanehbandi ₅ , Soudabe Yousefi ₁ , Masoud Darabi _{1,

2} , Reza Rahbarghazi _{2,

6}

Affiliation

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.

中文翻译：

花生四烯酸减轻了乙酰水杨酸对人颗粒细胞体外性能的有害影响。

在这里，我们研究了花生四烯酸（AA）在暴露于ASA后对人类卵丘颗粒细胞（CGC）的生物学作用。从卵泡液中分离细胞，并与0.5 mM乙酰水杨酸（ASA）和50 µM AA孵育。细胞存活力通过3-（4,5-二甲基噻唑-2-基）-2-5-二苯基溴化四氮唑测定进行分析。通过化学发光测定法测量E ₂和P ₄水平。CYP19A1，FACN和SCD1等基因的表达通过实时聚合酶链反应测定法测定。通过监测谷胱甘肽过氧化物酶活性来分析氧化状态。通过气相色谱技术分析脂肪酸谱。酶联免疫吸附法用于检测ASA和AA后CGC中的前列腺素E ₂（PGE ₂）。通过免疫荧光染色研究雌激素受体的蛋白质水平。超微结构变化通过透射电子显微镜成像进行评估。ASA处理可降低CGC中的E ₂产生，Cyp19a1表达，谷胱甘肽过氧化物酶（GPx）活性和雌二醇受体表达。添加AA阻止了ASA诱导的E ₂降低（p <.05）和Cyp19a1的表达。此外，AA通过促进GPx活性提高了暴露于ASA的CGC的抗氧化能力（p <.05）。与ASA组相比，AA增加了单不饱和脂肪酸/饱和脂肪酸的比例（p <.05）。AA的添加触发了ASA处理的CGCS中PGE ₂的合成和分泌（p <.05）。在ASA组中观察到细胞质空泡化，并用AA治疗增加了空泡化率。补充AA后雌激素受体的表达增加。数据表明，AA降低了72小时后ASA对人CGC的有害作用。

更新日期：2020-04-08

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