Jawad Ali, Oh Dongjin, Choi Hyerin, Kim Mirae, Ham Jaehyung, Oh Byoung Chol, Lee Joohyeong, Hyun Sang-Hwan
Veterinary Medical Center and College of Veterinary Medicine, Laboratory of Veterinary Embryology and Biotechnology (VETEMBIO), Chungbuk National University, Cheongju, Republic of Korea.
Institute of Stem Cell and Regenerative Medicine (ISCRM), Chungbuk National University, Cheongju, Republic of Korea.
Front Vet Sci. 2024 Dec 13;11:1475329. doi: 10.3389/fvets.2024.1475329. eCollection 2024.
Myo-inositol (Myo-Ins), the most abundant form of inositol, is an antioxidant and plays a crucial role in the development and reproduction of mammals and humans. However, information elucidating the role of Myo-Ins in porcine embryonic development after parthenogenetic activation (PA) is still lacking. Therefore, we investigated the effect of Myo-Ins on porcine embryos and its underlying mechanisms.
In this study, various concentrations of Myo-Ins (0, 5, 10, and 20 mM) were added to the porcine zygotic medium (PZM3) during the culture (IVC) of porcine embryos. Several characteristics were evaluated, including cleavage rate, blastocyst formation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels in 4-5 cell stage embryos, total cell number, apoptotic rate in blastocysts, mitochondrial membrane potential (MMP), mitochondrial quantity, mitochondrial stress in the blastocysts, and gene expression for antioxidant and mitochondrial function markers. Additionally, the immunofluorescence of was assessed.
The results showed that Myo-Ins at concentrations of 10 and 20 mM significantly increased the blastocyst formation rate compared to the control group. Embryos supplemented with 20 mM Myo-Ins exhibited higher GSH levels and lower ROS levels than those in the control group. Myo-Ins supplementation also decreased the rate of apoptosis and the apoptotic index in the treatment groups. Additionally, embryos supplemented with 20 mM Myo-Ins showed increased mitochondrial membrane potential (MMP), greater mitochondrial quantity, and reduced oxidative stress in the mitochondria. Interestingly, the expression levels of genes related to mitochondrial function and the nuclear erythroid factor 2-related factor () pathway were elevated in the Myo-Ins treated groups. Furthermore, immunofluorescence results indicated that 20 mM Myo-Ins significantly increased expression in blastocysts compared to the control group.
In conclusion, 20 mM Myo-Ins supplementation enhanced blastocyst development and improved mitochondrial function by regulating apoptosis, reducing oxidative stress, and activating the pathway.
肌醇(Myo-Ins)是最丰富的肌醇形式,是一种抗氧化剂,在哺乳动物和人类的发育与繁殖中起着关键作用。然而,关于Myo-Ins在孤雌激活(PA)后猪胚胎发育中作用的信息仍然缺乏。因此,我们研究了Myo-Ins对猪胚胎的影响及其潜在机制。
在本研究中,在猪胚胎体外培养(IVC)期间,向猪合子培养基(PZM3)中添加不同浓度的Myo-Ins(0、5、10和20 mM)。评估了几个特征,包括4-5细胞期胚胎的卵裂率、囊胚形成率、细胞内谷胱甘肽(GSH)和活性氧(ROS)水平、总细胞数、囊胚中的凋亡率、线粒体膜电位(MMP)、线粒体数量、囊胚中的线粒体应激以及抗氧化和线粒体功能标志物的基因表达。此外,还评估了免疫荧光。
结果表明,与对照组相比,10和20 mM浓度的Myo-Ins显著提高了囊胚形成率。添加20 mM Myo-Ins的胚胎比对照组表现出更高的GSH水平和更低的ROS水平。补充Myo-Ins还降低了治疗组的凋亡率和凋亡指数。此外,添加20 mM Myo-Ins的胚胎显示出线粒体膜电位(MMP)增加、线粒体数量增多以及线粒体氧化应激降低。有趣的是,在Myo-Ins处理组中,与线粒体功能和核红细胞因子2相关因子()途径相关的基因表达水平升高。此外,免疫荧光结果表明,与对照组相比,20 mM Myo-Ins显著增加了囊胚中的表达。
总之,补充20 mM Myo-Ins通过调节凋亡、降低氧化应激和激活途径,增强了囊胚发育并改善了线粒体功能。