Effect of Rosemarinic Acid on the Cryopreservation of Dairy Goats (Capra hircus) Semen
ZHANG Pei1, XU Gang-Gang1, YUAN Yuan2, ZHANG Jian-Yong3, YAN Tian-Rong4, PENG Zhong-Hong5, LI Zong-Ru6, HOU Jin-Xing7, AN Xiao-Peng1*
1 College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China; 2 School of Basic Medical Sciences, Air Force Military Medical University, Xi'an 710032, China; 3 Animal Disease Prevention and Control Center of Qian County, Xianyang 713300, China; 4 Heshui County Animal Disease Prevention and Control Center, Heshui 745400, China; 5 Shaanxi Ankang Animal Husbandry and Veterinary Medical Center, Ankang 725000, China; 6 Gansu Heshui County Agricultural Industry Development and Investment Co., Heshui 745400, China; 7 College of Animal Engineering, Yangling Vocational & Technical College, Yangling 712100, China
Abstract：Sperm cells are highly susceptible to damage during semen cryopreservation, in which excessive free radicals cause oxidative stress that negatively affects the cellular system by increasing the production of reactive oxygen species. Therefore, the use of antioxidants in semen cryopreservation is crucial. Rosmarinic acid (RA) is a natural polyphenolic antioxidant with strong antioxidant activity. In this study, 0, 20, 40, 80 and 120 μmol/L RA was added to the semen cryopreservation diluent of dairy goats (Capra hircus) to test the sperm quality and sperm antioxidant capacity, and sperm plasma metabolic profiles of the spermatozoa of dairy goats were analyzed by LC-MS non-targeted metabolomics after freezing. The results showed that RA improved the quality of cryopreserved dairy goat spermatozoa and sperm antioxidant capacity, and 309 differential metabolites were detected between the control group and RA-treated group. Moreover, RA regulated the energy metabolism of spermatozoa through AMP-activated protein kinase (AMPK) signaling pathway, and activated or inhibited apoptosis-related proteins, such as B-cell lymphoma-2 (Bcl-2), through PI3K-Akt signaling pathway. The present study demonstrated that the addition of 40 μmol/L RA could improve the effect of semen cryopreservation in dairy goats. This study provides a new idea and theoretical basis for the optimization of semen cryopreservation technology in dairy goats.
[1] 陈美杰. 2019. 丹参素和迷迭香酸对猪精液常温保存效果的研究[D]. 硕士学位论文, 西北农林科技大学, 导师: 胡建宏, pp. 8-76.(Chen M J.2011. Study on the effect of tanshin and rosemarinic acid on the preservation of porcine semen at room temperature[D]. Thesis for M.S., Northwest Agriculture and Forestry University, Supervisor: Hu J H, pp. 8-76.). [2] 霍浩然, 秦瑞峰, 薛佳栋, 等. 2022. 基于PI3K/Akt/mTOR通路探讨川芎嗪诱导人胃癌细胞凋亡和自噬作用的机制研究[J]. 西部中医药, 35(10): 12-16.(Huo H R, Qin R F, Xue J D, et al.2022. Mechanism of apoptosis and autophagy induced by chuanxiongxizine in human gastric cancer cells based on PI3K/Akt/mTOR pathway[J]. Western Traditional Chinese Medicine, 35(10): 12-16.) [3] 张星, 任发, 冯天雨, 等. 2023. 黄芪多糖对奶山羊精液冷冻保存效果的影响[J]. 家畜生态学报, (04): 66-73.(Zhang X, Ren F, Feng T Y, et al. 2023. Effects of astragalus polysaccharide on the cryopreservation of semen in dairy goats[J]. Journal of Livestock Ecology, (04): 66-73.) [4] Al Danaf N, Melhem R A, Assaf K I, et al.2016. Photophysical properties of neutral and dissociated forms of rosmarinic acid[J]. Journal of Luminescence, 175: 50-56. [5] Barri T,Dragsted L O.2013. UPLC-ESI-QTOF/MS and multivariate data analysis for blood plasma and serum metabolomics: Effect of experimental artefacts and anticoagulant[J]. Analytica Chimica Acta, 768: 118-128. [6] Bezerra I M A, Souza J A T, Sousa Junior A, et al.2022. Effect of fortification of extender with alpha-terpineol and rosemary essential oil on post-thaw ovine semsen quality[J]. Reproduction in Domestic Animals, 57(9): 1056-1062. [7] Evans E P P, Scholten J T M, Mzyk A, et al.2021. Male subfertility and oxidative stress[J]. Redox Biology, 46: 102071. [8] Feng T Y, Lv D L, Zhang X, et al.2020. Rosmarinic acid improves boar sperm quality, antioxidant capacity and energy metabolism at 17 degrees C via AMPK activation[J]. Reproduction in Domestic Animals, 55(12): 1714-1724. [9] Fernandez M C, Yu A, Moawad A R, et al.2019. Peroxiredoxin 6 regulates the phosphoinositide 3-kinase/AKT pathway to maintain human sperm viability[J]. Molecular Human Reproduction, 25(12): 787-796. [10] Fisher A B, Dodia C, Sorokina E M, et al.2016. A novel lysophosphatidylcholine acyl transferase activity is expressed by peroxiredoxin 6[J]. Journal of Lipid Research, 57(4): 587-596. [11] He Y L, Li D, Zhang W Y, et al.2020. Boar sperm quality and oxidative status as affected by rosmarinic acid at 17 ℃[J]. Tropical Animal Health and Production, 52(4): 2169-2177. [12] Hu S G, Liang A J, Yao G X, et al.2018. The dynamic metabolomic changes throughout mouse epididymal lumen fluid potentially contribute to sperm maturation[J]. Andrology, 6(1): 247-255. [13] Huang Q, Liu L, Wu Y, et al.2019. Seminal plasma metabolites mediate the associations of multiple environmental pollutants with semen quality in Chinese men[J]. Environment International, 132: 105066. [14] Jang H J, Yang K E, Oh W K, et al.2019. Nectandrin B-mediated activation of the AMPK pathway prevents cellular senescence in human diploid fibroblasts by reducing intracellular ROS levels[J]. Aging, 11(11): 3731-3749. [15] Kim G D, Park Y S, Jin Y H, et al.2015. Production and applications of rosmarinic acid and structurally related compounds[J]. Applied Microbiology and Biotechnology, 99(5): 2083-92. [16] Kutluyer F, Aksu O, Kocabas M.2019. Effect of L-Tryptophan on sperm quality of tigris scraper (Capoeta umbla) (Pisces: Cyprinidae) after cryopreservation[J]. Cryo Letters, 40(2): 77-82. [17] Li Z, Song Y, Hou W, et al.2022. Atractylodin induces oxidative stress-mediated apoptosis and autophagy in human breast cancer MCF-7 cells through inhibition of the P13K/Akt/mTOR pathway[J]. Journal of Biochemical and Molecular Toxicology, 36(8): e23081. [18] Lv M G, Chen W Q, Weng S Q, et al.2018. Rosmarinic acid compromises human sperm functions by an intracellular Ca2+ concentration-related mechanism[J]. Reproductive Toxicology, 81: 58-63. [19] Mercati F, Domingo P, Pasquariello R, et al.2020. Effect of chelating and antioxidant agents on morphology and DNA methylation in freeze-drying rabbit (Oryctolagus cuniculus) spermatozoa[J]. Reproduction in Domestic Animals, 55(1): 29-37. [20] Salmon V M, Leclerc P,Bailey J L.2016. Cholesterol-loaded cyclodextrin increases the cholesterol content of goat sperm to improve cold and osmotic resistance and maintain sperm function after cryopreservation[J]. Biology of Reproduction, 94(4): 85. [21] Shan S, Xu F, Hirschfeld M, et al.2021. Sperm lipid markers of male fertility in mammals[J]. International Journal of Molecular Sciences, 22(16): 8767. [22] Silvestre M A, Yaniz J L, Pena F J, et al.2021. Role of antioxidants in cooled liquid storage of mammal spermatozoa[J]. Antioxidants (Basel), 10(7): 1096. [23] Sun L, Fan X, Zeng Y, et al.2020. Resveratrol protects boar sperm in vitro via its antioxidant capacity[J]. Zygote(Cambridge, England): 1-8. [24] Touazi L, Aberkane B, Bellik Y, et al.2018. Effect of the essential oil of Rosmarinus officinalis (L.) on rooster sperm motility during 4 degrees C short-term storage[J]. Veterinary World, 11(5): 590-597. [25] Wang Y, Meng J, Men L, et al.2020. Rosmarinic acid protects mice from concanavalin a-induced hepatic injury through AMPK signaling[J]. Biological & Pharmaceutical Bulletin, 43(11): 1749-1759. [26] Want E J, Masson P, Michopoulos F, et al.2013. Global metabolic profiling of animal and human tissues via UPLC-MS[J]. Nature Protocols, 8(1): 17-32. [27] Yeni D, Inanc M E, Avdatek F, et al.2018. Supplementation of rosmarinic acid has reduced oxidative stress on bull spermatozoa following the freeze thawing process[J]. Cryo Letters, 39(2): 156-165. [28] Zhang Q, Song X, Chen X, et al.2021. Antiosteoporotic effect of hesperidin against ovariectomy-induced osteoporosis in rats via reduction of oxidative stress and inflammation[J]. Journal of Biochemical and Molecular Toxicology, 35(8): e22832. [29] Zidni I, Lee H B, Yoon J H, et al.2022. Effect of antioxidants in cryopreservation media on spotted halibut (Verasfer variegatus) sperm quality during cryopreservation[J]. Aquaculture, 557: 738351. [30] Zou J H, Wei L X, Li D X, et al.2021. Effect of glutathione on sperm quality in guanzhong dairy goat sperm during cryopreservation[J]. Frontiers in Veterinary Science, 8: 771440.
