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辅酶 Q10 和姜黄素补充剂对人精子冷冻保存液的影响。

The effects of coenzyme Q10 and curcumin supplementation in freezing medium for human sperm cryopreservation.

机构信息

Ankara City Hospital, Center for Assisted Reproduction, Obstetrics and Gynecology Clinics, Ankara, Turkey.

University of Dundee, School of Medicine, Scotland, UK.

出版信息

Eur J Obstet Gynecol Reprod Biol. 2023 Aug;287:36-45. doi: 10.1016/j.ejogrb.2023.05.040. Epub 2023 Jun 2.

Abstract

BACKGROUND

Despite its routine and frequent application, cryopreservation of human sperm is far from the desired efficacy, as freezing and thawing impair motility, viability, acrosomal unity, and DNA integrity.

OBJECTIVES

In this study, the authors aimed to investigate whether adding antioxidants, coenzyme Q10, and curcumin into the freezing medium provide better efficacy in the cryopreservation of human sperm.

METHODS

The semen samples from 40 healthy men aged 18-45 were collected in sterile containers by masturbation. Samples within normal reference values for sperm concentration (≥15 million/mL) and motility (progressive motile ≥ 32% and total motility ≥ 40%) were included in the study. Semen samples were equally divided into five groups and evaluated; i) pre-freezing sperm suspension, ii) frozen-thawed control (Ctrl) without any supplementation in freezing medium, iii) frozen-thawed with curcumin supplementation of 0.25 mM (Cur), iv) frozen-thawed coenzyme Q10 supplementation of 25 µM (CoQ10) and v) frozen-thawed curcumin (0.25 mM) plus coenzyme Q10 (25 µM) supplementation (CurCoQ10) into the freezing medium. Liquid nitrogen vapour freezing and rapid thawing were performed in each group (ii-v). Sperm motility, viability, acrosome integrity, and DNA fragmentation rates were compared and ultrastructural evaluations by transmission electron microscopy were undertaken between the groups. Additionally, the total antioxidant capacity/total oxidant capacity values were measured.

RESULTS

According to CASA results, progressive motility was significantly higher in the CoQ10 group (9.4 ± 7.6) when compared with the Ctrl (7.1 ± 6.3), Cur (6.4 ± 4.8) and CurCoQ10 (8.1 ± 7.7) groups (p < 0.05). Flow cytometry results showed no difference in the viability and acrosome integrity values after thawing, but DNA fragmentation was significantly increased in the curcumin-added groups (p < 0.05). Acrosomal changes and sub-acrosomal defects were seen in all groups after thawing at the ultrastructural level. Mitochondrial membrane structure was preserved in CoQ10 and CurCoQ10 groups.

CONCLUSIONS

Our results suggested that sperm ultrastructural morphology and motility were better preserved in the CoQ10 group during cryopreservation. In curcumin groups, DNA fragmentation and head defects were increased.

摘要

背景

尽管冷冻保存人类精子是一种常规且频繁应用的技术,但冷冻和解冻过程会损害精子的运动能力、活力、顶体完整性和 DNA 完整性,因此其效果远非理想。

目的

本研究旨在探讨在冷冻液中添加抗氧化剂辅酶 Q10 和姜黄素是否能提高人类精子的冷冻效果。

方法

本研究采集了 40 名年龄在 18-45 岁之间的健康男性的精液样本,采集方法为自慰。纳入的样本精子浓度(≥1500 万/ml)和活力(前向运动精子≥32%和总活力≥40%)均在正常参考范围内。将精液样本等分为五组进行评估:i)冷冻前精子悬液,ii)未添加任何物质的冷冻-解冻对照组(Ctrl),iii)添加 0.25mM 姜黄素的冷冻-解冻组(Cur),iv)添加 25µM 辅酶 Q10 的冷冻-解冻组(CoQ10),v)添加 0.25mM 姜黄素和 25µM 辅酶 Q10 的冷冻-解冻组(CurCoQ10)。每组均进行液氮蒸气冷冻和快速解冻。比较各组精子运动能力、活力、顶体完整性和 DNA 碎片化率,并通过透射电子显微镜进行超微结构评估。此外,还测量了总抗氧化能力/总氧化能力值。

结果

根据 CASA 结果,与 Ctrl、Cur 和 CurCoQ10 组相比,CoQ10 组解冻后的前向运动精子活力(9.4±7.6)明显更高(p<0.05)。流式细胞术结果显示,解冻后精子的活力和顶体完整性值无差异,但姜黄素添加组的 DNA 碎片化明显增加(p<0.05)。超微结构水平显示,解冻后所有组均出现顶体改变和亚顶体缺陷。CoQ10 和 CurCoQ10 组保留了线粒体膜结构。

结论

本研究结果表明,在冷冻保存过程中,CoQ10 组精子的超微结构形态和运动能力得到更好的保存。在姜黄素组中,DNA 碎片化和头部缺陷增加。

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