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Effect of zinc chloride and sodium selenite supplementation on in vitro maturation, oxidative biomarkers, and gene expression in buffalo (Bubalus bubalis) oocytes
Zygote ( IF 1.7 ) Pub Date : 2021-03-26 , DOI: 10.1017/s0967199421000162
Wael A Khalil 1 , Chun-Yan Yang 2 , Mostafa M El-Moghazy 3 , Mohamed S El-Rais 3 , Jiang-Hua Shang 2 , Ashraf El-Sayed 4, 5
Affiliation  

SummaryThis study examined the effects of zinc chloride (ZnCl2) and sodium selenite (Na2SeO3) supplementation in maturation medium on in vitro maturation (IVM) rate, oxidative biomarkers and gene expression in buffalo oocytes. Ovaries from a slaughterhouse were aspirated and good quality cumulus–oocyte complexes (COCs) with at least four layers of compact cumulus cells and evenly granulated dark ooplasm were selected. COCs were randomly allocated during IVM (22 h) to one of four treatment groups: (1) control maturation medium (basic medium), or basic medium supplemented with (2) ZnCl2 (1.5 µg/ml), (3) Na2SeO3 (5 µg/l), or (4) ZnCl2 + Na2SeO3 (1.5 µg/ml + 5 µg/l, respectively). Oocytes were denuded after 22 h of IVM in the first four replicates. Specimens were fixed and stained to evaluate the stage of nuclear maturation. The spent medium was collected for biochemical assays of total antioxidant capacity (TAC), malondialdehyde (MDA) and hydrogen peroxide concentrations. A second four replicates were used for COCs for RNA extraction. The expression levels of antioxidant (SOD1, GPX4, CAT and PRDX1), antiapoptotic (BCL2 and BCL-XL) and proapoptotic (BAX and BID) genes were measured. Supplementation with ZnCl2 and Na2SeO3 during IVM increased the ratio of oocytes reaching metaphase II at 22 h, increased TAC and decreased MDA and H2O2 concentrations in the maturation medium (P < 0.05). Moreover, beneficial effects were associated with complementary changes in expression patterns of antioxidative, antiapoptotic and proapoptotic genes, suggesting lower oxidative stress and apoptosis. Supplementation medium with zinc chloride and sodium selenite improves the maturation rate, reduces oxidative stress and increases expression levels of antioxidative and antiapoptotic genes.

中文翻译:

补充氯化锌和亚硒酸钠对水牛(Bubalus bubalis)卵母细胞体外成熟、氧化生物标志物和基因表达的影响

摘要本研究检验了氯化锌(ZnCl2) 和亚硒酸钠 (Na2硒化3) 在成熟培养基中补充体外水牛卵母细胞的成熟 (IVM) 率、氧化生物标志物和基因表达。抽取来自屠宰场的卵巢并选择具有至少四层致密卵丘细胞和均匀颗粒状深色卵质的优质卵丘-卵母细胞复合物 (COC)。在 IVM(22 小时)期间将 COC 随机分配到四个治疗组之一:(1)对照成熟培养基(基础培养基),或补充有(2)氯化锌的基础培养基2(1.5 µg/ml), (3) 钠2硒化3(5 µg/l),或 (4) 氯化锌2+ 钠2硒化3(分别为 1.5 µg/ml + 5 µg/l)。在前四次重复中,IVM 22 小时后卵母细胞被剥光。标本被固定和染色以评估核成熟的阶段。收集用过的培养基用于总抗氧化能力 (TAC)、丙二醛 (MDA) 和过氧化氢浓度的生化测定。第二个四次重复用于 COC,用于 RNA 提取。抗氧化剂的表达水平(SOD1,GPX4,PRDX1), 抗凋亡 (BCL2BCL-XL) 和促凋亡 (BAX出价) 基因进行了测量。补充氯化锌2和钠2硒化3在 IVM 期间,卵母细胞在 22 小时达到中期 II 的比例增加,TAC 增加,MDA 和 H 降低22成熟培养基中的浓度(磷 <0.05)。此外,有益作用与抗氧化、抗凋亡和促凋亡基因表达模式的互补变化有关,表明氧化应激和细胞凋亡较低。含有氯化锌和亚硒酸钠的补充培养基提高了成熟率,减少了氧化应激并增加了抗氧化和抗凋亡基因的表达水平。
更新日期:2021-03-26
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