益母草碱对绵羊精液冷冻保存的影响

doi:10.3969/j.issn.1674-7968.2023.10.011

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摘要精液冷冻保存是有效提升绵羊(Ovis aries)人工授精效率的有效手段之一,在精液保存稀释液中添加抗氧化剂是精液保存过程的关键,益母草碱作为一种抗氧化剂在精液保存的研究中鲜有报告。本研究在多浪羊精液冷冻保存稀释液中分别添加20、40、60、80和100 μmol/mL的益母草碱,通过检测精液品质、抗氧化能力、相关凋亡基因mRNA相对表达量及受胎率等指标,综合评价益母草碱对绵羊精液冷冻保存效果的影响。研究结果表明,与对照组(0 μmol/mL)相比,在冷冻保存稀释液中添加益母草碱后,能显著提高多浪羊冷冻保存精液的活率、顶体完整率和质膜完整率(P<0.05);并显著提升精液超氧化物歧化酶(superoxide dismutase, SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-PX)和过氧化氢酶(catalase, CAT)的活性,提高总抗氧化能力(total antioxidant eapacity, T-AOC)水平并降低丙二醛(malondialdehyde, MDA)的含量(P<0.05);其中,冷冻保存液中益母草碱添加浓度为60 μmol/mL时保存效果最佳(P<0.05);此外,与对照组相比,在冷冻保存过程中,添加最佳浓度益母草碱组的精子相关凋亡基因肿瘤坏死因子受体超族成员6 (TNF receptor superfamily, member 6, Fas)、半胱天冬酶家族-3 (caspase family omega-3, Caspase-3)和B淋巴细胞瘤-2关联X蛋白(Bcl2-associated X, Bax)的mRNA相对表达量显著降低(P<0.05),抗凋亡基因B淋巴细胞瘤-2 (B-cell lymphoma-2, Bcl-2)相对表达量显著升高(P<0.05);同时冷冻保存液中添加浓度为60 μmol/mL组精子的受胎率为60%,高于0 μmol/mL组,达到鲜精阴道输精的最低水平。本研究结果表明,在精液稀释液中添加浓度为60 μmol/mL的益母草碱后可显著改善多浪羊冷冻保存精液品质,提升其抗氧化能力,同时起到抗凋亡作用,使受胎率达到一个良好的水平。本研究为探索益母草碱对提高绵羊精液保存效果提供了有益借鉴。

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	李智星
	卿育洪
	李志远
	曹少奇
	高新梅
	曾维斌
	王静
	胡广东

关键词 ：精液保存, 益母草碱, 精液品质, 抗氧化能力, 抗凋亡能力, 受胎率

Abstract：Cryopreservation of semen is one of the effective means to effectively improve the efficiency of artificial insemination in Ovis aries. Adding antioxidants to semen preservation diluent is the key to semen preservation process. Leonurine alkaloid as an antioxidant, has rarely been reported in semen preservation studies. In this study, 20, 40, 60, 80 and 100 μmol/mL leonurine alkaloid were added to the cryopreserved diluent of Duolang sheep semen, respectively. The effect of leonurine alkaloid on the cryopreservation of sheep semen was comprehensively evaluated by detecting semen quality, antioxidant capacity, relative expression of apoptotic related gene mRNA, and conception rate. The results showed that compared with the control group (0 μmol/mL), the supplementation of leonurine alkaloid in the cryoppreservation diluent could significantly increase the viability rate, acrosome integrity rate and plasma membrane integrity rate of the cryoppreservation semen of Duolang sheep (P<0.05); It significantly increased the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and catalase (CAT), increased the level of total antioxidant eapacity (T-AOC) and decreased the content of malondialdehyde (MDA)(P<0.05); The best preservation effect of leonurine alkaloid in cryopreservation was 60 μmol/mL (P<0.05); In addition, compared with the 0 μmol/mL group, the relative mRNA expressions of spermatozoa-related apoptotic genes TNF receptor superfamily, member 6 (Fas), caspase family omega-3 (Caspase-3) and BCL2-associated X (Bax) were significantly decreased (P<0.05), and the relative expression of anti-apoptotic gene B-cell lymphoma-2 (Bcl-2) was significantly increased in the optimal concentration of leonurine alkaloid group during cryopreserved (P<0.05); At the same time, the conception rate of 60 μmol/mL group was higher than that of 0 μmol/mL group, which reached the lowest level of fresh spermatozoa vaginal instillation. This study showed that adding 60 μmol /mL leonurine alkaloid to semen diluent could significantly improve the quality of frozen preserved semen of Duolang sheep, enhance its antioxidant capacity, and play an anti-apoptotic role, so that the conception rate could reach a good level. This study provides a useful reference for exploring the effect of leonurine alkaloid on sheep semen preservation and improving the effect of sheep semen preservation.

Key words： Semen preservation Leonurine alkaloid Semen quality Antioxidant capacity Antiapoptotic ability Conception rate

收稿日期: 2022-10-06

ZTFLH:

S814.3

基金资助:兵团重点领域科技攻关计划项目(2021AB014); 石河子大学创新发展专项(CXFZ202112); 自治区多浪羊品种选育推广技术体系专项(2021DLY-23)

引用本文:

李智星, 卿育洪, 李志远, 曹少奇, 高新梅, 曾维斌, 王静, 胡广东. 益母草碱对绵羊精液冷冻保存的影响[J]. 农业生物技术学报, 2023, 31(10): 2115-2122.
LI Zhi-Xing, QING Yu-Hong, LI Zhi-Yuan, CAO Shao-Qi, GAO Xin-Mei, ZENG Wei-Bin, WANG Jing, HU Guang-Dong. The Effect of Leonurine on Cryopreservation of Sheep (Ovis aries) Semen. 农业生物技术学报, 2023, 31(10): 2115-2122.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2023.10.011 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2023/V31/I10/2115

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