奶牛卵泡颗粒细胞在卵泡生长过程中的氧化过程

Yulia Bodnar, N. Kuzmina, D. Ostapiv, O. I. Chajkovska, R. Ostapiv
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引用次数: 0

摘要

本文介绍了在卵泡生长生理状态下,奶牛卵泡颗粒层细胞呼吸活性和氧化还原能力的研究结果。屠宰后选择卵泡生长生理状态的卵巢,评价卵泡大小。从小(< 4 mm)、中(4 - 7 mm)和大(> 7 mm)卵泡中抽取颗粒层细胞,极谱法测定其吸氧强度,电位法测定其氧化还原能力。K3[Fe(CN)6]作为胞外电子受体。采用氟化钠抑制剂、线粒体电子传递链的nadd依赖环节- amytal和末端(细胞色素氧化酶)-叠氮化钠来测定有氧糖酵解所消耗的氧份额。用Na2EDTA法测定了不饱和脂肪酸自由基氧化在氧生成过程中所占的份额。卵泡生长的卵巢颗粒细胞具有呼吸活动和再生能力,其强度取决于提取它们的卵泡的大小。所研究的指标的最高值是取自大卵泡的细胞的特征,较低的是取自小卵泡,最低的是取自中等卵泡。来自小卵泡的颗粒中的高强度氧化过程确保了卵母细胞的生长,而来自大卵泡的优势卵泡中的生殖细胞成熟。同时,小毛囊和显性毛囊细胞的耗氧强度增加是在细胞外环境中电子受体限制的背景下表现出来的。糖酵解过程中ATP再合成抑制剂的使用和线粒体呼吸链单个环节的使用证明了颗粒中所指示的代谢途径的活性,以及使用替代底物以确保细胞存在的可能性。小毛囊和大毛囊颗粒层细胞利用氧气的主要方式是有氧糖酵解,从培养基中也有供氧和利用底物的替代方式。由于糖酵解的抑制,中等大小的卵泡颗粒中还原性等效物流入细胞外空间的量最大,这表明进入卵泡腔的还原性等效物的来源是糖酵解,这表明细胞的存在依赖于所指示的代谢途径。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
OXIDATIVE PROCESSES IN GRANULOSA CELLS OF COW OVARIAN FOLLICLES AT FOLLICULAR GROWTH
Article presents the research results on the respiratory activity and redox capacity of granulose layer cells of cow ovarian follicles under the physiological state of follicular growth. After the slaughter of the cows, the ovaries in physiological state of follicular growth were selected and the size of the follicles was evaluated. Cells of the granulosa layer were aspirated from small (< 4 mm), medium (4–7 mm) and large (> 7 mm) follicles and determined: intensity of oxygen absorption – polarographically, redox capacity – potentiometrically. K3[Fe(CN)6] was used as an extracellular electron acceptor. The share of oxygen consumed by aerobic glycolysis was determined by using sodium fluoride inhibitor, the NAD-dependent link of the mitochondrial electron transport chain - amytal, and the terminal (cytochrome oxidase) – sodium azide. The share of free-radical oxidation of unsaturated fatty acids in the realization of oxygen was detected by Na2EDTA. Granulosa cells from the ovary of follicular growth are characterized by respiratory activity and regenerative capacity, the intensity of which depends on the size of the follicles from which they are extracted. The highest values of the investigated indicators are characteristic of cells extracted from large follicles, lower – from small follicles, and the lowest values are from medium follicles. The high intensity of oxidative processes in granulosa from small follicles ensures the growth of oocytes, and from large follicles – maturation of the germ cell in the dominant follicle. At the same time, the increased intensity of oxygen consumption by the cells of small and dominant follicles is manifested against the background of limitations in electron acceptors in the extracellular environment. The use of inhibitors of ATP resynthesis of both glycolysis and individual links of the mitochondrial respiratory chain testify the activity of the indicated metabolic pathways in granulosa and the possibility of using alternative substrates to ensure the existence of cells. The main way of utilization of oxygen in the granulosa layer cells of small and large follicles is aerobic glycolysis, and from the medium there are alternative ways of supplying and using substrates. The largest flow of reducing equivalents into the extracellular space from the granulosa of a medium-sized follicle due to inhibition of glycolysis indicates that the source of the supply of reducing equivalents into the follicle cavity is glycolysis and this characterizes the dependence of the existence of cells on the indicated metabolic pathway.
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