益母草碱对绵羊精液低温保存效果的影响

doi:10.16431/j.cnki.1671-7236.2023.07.020

Abstract

Abstract: 【Objective】 This study was aimed to explore the effect of leonurine on the cryopreservation effect of sheep semen, and provide theoretical basis for the preparation of appropriate diluent for cryopreservation of sheep semen in practical production.【Method】 Semen of three Duolang sheep were collected by pseudoginosis method.0 (control), 20, 40, 60, 80 and 100 μmol/mL leonurine was added to semen cryopreserved diluent, respectively.Semen quality indexes such as sperm viability, acrosome integrity and plasma membrane integrity, total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) activities, and malondialdehyde (MDA) content were detected during semen cryoppreservation.The optimal concentration of leonuri during semen preservation was screened.After the optimal concentration of leonuri was added, the expression of TNF receptor superfamily member 6 (Fas), Caspase-3, B lymphoblastoma-2 (Bcl-2) and B lymphoblastoma-2-associated X protein (Bax) during cryopreserved semen were detected by Real-time quantitative PCR.The conception rate of ewe was calculated using the method of two intraventricular insemination.【Result】 Compared with control group, the sperm viability, plasma membrane integrity, acrosome integrity, T-AOC level, SOD, GSH-Px and CAT activities of 40 μmol/mL leonurine group were significantly increased (P<0.05), while the content of MDA was significantly decreased (P<0.05). Cryopreservation for 144 h, the indexes of 40 μmol/mL leonurine group were significantly different from those of other leonurine groups (P<0.05).Real-time quantitative PCR results showed that compared with control group, the expressions of Fas, Caspase-3 and Bax genes in 40 μmol/mL leonurine group were significantly decreased (P <0.05), while the expression of Bcl-2 gene was significantly increased (P<0.05).【Conclusion】 In this study, adding 40 μmol/mL leonurine to cryopreservation semen diluent could significantly improve the quality of cryopreservation of Duolang sheep semen, enhance the antioxidant capacity, and played an anti-apoptotic role.The conception rate of ewes with 40 μmol/mL leonurine cryopreservation for 72 h was 75%, which reached the average level of vaginal transfusion of fresh semen.

Key words: leonurine; cryopreservation; semen quality; antioxidant capacity; conception rate

CLC Number:

S814.3

LI Zhixing, QING Yuhong, LI Zhiyuan, CAO Shaoqi, GAO Xinmei, ZENG Weibin, WANG Jing, HU Guangdong. Effects of Leonurine on Cryopreservation of Sheep Semen[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(7): 2801-2810.

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Effects of Leonurine on Cryopreservation of Sheep Semen

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