Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nitra, Slovakia.
Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nitra, Slovakia.
Toxicol In Vitro. 2024 Aug;99:105855. doi: 10.1016/j.tiv.2024.105855. Epub 2024 May 28.
Epigenetic methods to prevent the reproductive toxicity of oil-related environmental contaminants are currently unavailable. The present study aimed to examine the ability of the microRNA miR-152 to mitigate the effects of benzene on ovarian cells. Porcine ovarian granulosa cells transfected or not transfected with miR-152 mimics were cultured with or without benzene (0, 10 and 100 ng/ml). The expression of miR-152; viability; proliferation (cell proliferation and expression of mRNAs and accumulation of PCNA and cyclin B1); apoptosis (expression of mRNAs and accumulation of bax and caspase 3; and the proportion of cells with fragmented DNA); and release of progesterone, estradiol and IGF-I were analyzed via RT-qPCR; the Trypan blue exclusion test; quantitative immunocytochemistry; BrdU; XTT; TUNEL assays; and ELISA. Administration of benzene promoted the expression of apoptosis markers and reduced cell viability, all measured markers of proliferation, the release of steroid hormones and IGF-I. Overexpression of miR-152 was associated with increased cell viability, proliferation, progesterone and IGF-I release and reduced apoptosis and estradiol output. Moreover, miR-152 mitigated or prevented the effects of benzene on all the measured parameters in addition to estradiol release. The present observations suggest the toxic effect of benzene and the stimulatory influence of miR-152 on ovarian cell functions. Moreover, this is the first demonstration of the ability of miRNAs to mitigate and prevent the reproductive toxicity of benzene.
目前尚无预防与油相关的环境污染物的生殖毒性的表观遗传方法。本研究旨在研究 microRNA miR-152 减轻苯对卵巢细胞影响的能力。用或不用 miR-152 模拟物转染的猪卵巢颗粒细胞在有或没有苯(0、10 和 100ng/ml)的情况下培养。分析 miR-152 的表达;活力;增殖(细胞增殖和 mRNAs 的表达以及 PCNA 和细胞周期蛋白 B1 的积累);凋亡(mRNAs 的表达以及 bax 和 caspase 3 的积累和具有片段化 DNA 的细胞的比例);以及孕激素、雌二醇和 IGF-I 的释放通过 RT-qPCR;台盼蓝排除试验;定量免疫细胞化学;BrdU;XTT;TUNEL 测定;和 ELISA。苯的给药促进了凋亡标志物的表达并降低了细胞活力,所有测量的增殖标志物、类固醇激素和 IGF-I 的释放。miR-152 的过表达与细胞活力、增殖、孕激素和 IGF-I 释放的增加以及凋亡和雌二醇产量的减少有关。此外,miR-152 减轻或预防了苯对所有测量参数的影响,除了雌二醇的释放。这些观察结果表明了苯的毒性作用以及 miR-152 对卵巢细胞功能的刺激作用。此外,这是首次证明 miRNA 能够减轻和预防苯的生殖毒性。