Incubating frozen-thawed buffalo sperm with olive fruit extracts counteracts thawing-induced oxidative stress and improves semen quality

IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY
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Abstract

Freezing-thawing procedures and semen manipulation for in vitro fertilization induce oxidative stress, which in turn leads to impaired sperm quality. The aim of this study was to evaluate whether incubation of frozen-thawed buffalo semen with olive fruit extracts (OFE), known to contain a high concentration of phenolic antioxidants, would improve semen quality by reducing oxidative stress.

Frozen sperm (4 ejaculates/4 bulls/3 replicates) were thawed and diluted to 30 × 106/mL in IVF medium with 0, 72, 143, and 214 μL/mL of OFE, corresponding to 0 (D0-control), 50 (D50), 100 (D100), and 150 (D150) μM hydroxytyrosol. Sperm viability, acrosome integrity, membrane functionality, motility, and sperm kinetics were evaluated immediately after thawing (T0) and after 1 (T1) and 2 h (T2) of incubation at 38.7 °C. Based on the results, sperm biological antioxidant potential (BAP) and ROS levels (ROMs) were assessed in D0 and D100 groups at T1 and T2. To assess the effect of OFE on fertilizing ability, heterologous penetration rates were also evaluated, using bovine abattoir-derived oocytes.

The treatment with OFE at all concentrations tested increased (P < 0.05) the percentage of acrosome intact spermatozoa compared to the D0-control at T1, but the effect was more evident (P < 0.01) with D100 (54.5 ± 3.0, 60.5 ± 1.5, 65.2 ± 3.3, and 62.5 ± 1.7, with D0, D50, D100, and D150 OFE, respectively). Total motility, progressive motility, rapid velocity, and progressive velocity decreased (P < 0.05) at T2 only in the D0-control group. The percentage of rapidly progressive sperm and the progressive motility tended to increase (P < 0.10) at T1 and T2, respectively, in D100 compared to D0 (24.7 ± 4.1 vs 16.4 ± 1.6 and 22.8 ± 2.7 vs 17.0 ± 1.2, respectively). The treatment with D100 OFE of frozen-thawed sperm increased (P < 0.05) some kinetic parameters (VAP and WOB). Spermatozoa incubated with D100 OFE exhibited higher (P < 0.01) total and normospermic oocyte penetration rates compared to D0 (86.5 ± 1.4 vs 78.5 ± 0.7, and 70.6 ± 1.5 vs 63.8 ± 1.1, respectively). Additionally, D100 OFE increased sperm BAP concentrations at both T1 and T2, while ROS levels were unaffected. These results suggest that incubating frozen-thawed buffalo semen with OFE is an effective strategy for preserving semen quality and in vitro fertilization ability by enhancing sperm antioxidant capacity.

用橄榄果提取物培养冷冻解冻的水牛精子,可抵消解冻引起的氧化应激,提高精液质量
用于体外受精的冷冻-解冻程序和精液处理会诱发氧化应激,进而导致精子质量受损。橄榄果提取物含有高浓度的酚类抗氧化剂,本研究旨在评估用橄榄果提取物培养冷冻解冻的水牛精液是否能通过降低氧化应激改善精液质量。将冷冻精子(4个射精/4头公牛/3个重复)解冻并在IVF培养基中稀释至30×106/毫升,分别加入0、72、143和214微升/毫升的橄榄果提取物,相当于0(D0-对照)、50(D50)、100(D100)和150(D150)微摩尔羟基酪醇。在解冻后立即(T0)以及在 38.7 °C 下培养 1 小时(T1)和 2 小时(T2)后,对精子的存活率、顶体完整性、膜功能、运动能力和精子动力学进行了评估。根据结果,在 T1 和 T2 时评估了 D0 和 D100 组精子的生物抗氧化潜能(BAP)和 ROS 水平(ROMs)。为了评估 OFE 对受精能力的影响,还使用牛屠宰场卵母细胞评估了异源穿透率。与 D0 对照组相比,用所有测试浓度的 OFE 处理 T1 时,顶体完好精子的百分比都会增加(P <0.05),但 D100 的影响更明显(P <0.01)(D0、D50、D100 和 D150 OFE 的影响分别为 54.5 ± 3.0、60.5 ± 1.5、65.2 ± 3.3 和 62.5 ± 1.7)。只有D0对照组的总运动能力、渐进运动能力、快速速度和渐进速度在T2下降(P <0.05)。与D0对照组相比,D100对照组在T1和T2的快速进展精子百分比和进展运动能力分别趋于增加(P < 0.10)(分别为24.7 ± 4.1 vs 16.4 ± 1.6和22.8 ± 2.7 vs 17.0 ± 1.2)。用 D100 OFE 处理冷冻解冻的精子会增加一些动力学参数(VAP 和 WOB)(P < 0.05)。与 D0 相比,用 D100 OFE 培养的精子表现出更高的(P < 0.01)总穿透率和正常精子卵母细胞穿透率(分别为 86.5 ± 1.4 vs 78.5 ± 0.7 和 70.6 ± 1.5 vs 63.8 ± 1.1)。此外,D100 OFE 还增加了精子在 T1 和 T2 的 BAP 浓度,而 ROS 水平则不受影响。这些结果表明,用 OFE 培养冷冻解冻的水牛精液是通过提高精子抗氧化能力来保持精液质量和体外受精能力的有效策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Theriogenology
Theriogenology 农林科学-生殖生物学
CiteScore
5.50
自引率
14.30%
发文量
387
审稿时长
72 days
期刊介绍: Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.
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