比较证据支持靶向线粒体的（Mitoquinone）抗氧化剂比细胞溶质的（Resveratrol）抗氧化剂在改善冷冻水牛（Bubalus bubalis）精液体外精子功能方面具有更好的抗氧化功效。
Comparative evidence support better antioxidant efficacy of mitochondrial-targeted (Mitoquinone) than cytosolic (Resveratrol) antioxidant in improving in-vitro sperm functions of cryopreserved buffalo (Bubalus bubalis) semen.
机构信息
Artificial Breeding Research Centre, LPM Division, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
Animal Reproduction, Gynaecology and Obstetrics, LPM Division, ICAR-National Dairy Research Institute, Karnal, Haryana, India.
出版信息
Cryobiology. 2021 Aug;101:125-134. doi: 10.1016/j.cryobiol.2021.04.007. Epub 2021 Apr 30.
The present study compared the effect of mitochondria-targeted (Mitoquinone, MitoQ) and untargeted cytosolic antioxidant (Resveratrol, RESV) supplementation on lipid peroxidation (LPO) and in-vitro sperm functions of cryopreserved buffalo bull semen. To optimize additive's concentration, sperm pellet obtained from twenty-four ejaculates was supplemented with different concentrations of MitoQ (20 nM, 100 nM, 200 nM); and RESV (10 μM, 25 μM, 50 μM) against control in the extender. The post-thaw sperm motility, livability, and membrane integrity were higher (P < 0.05) in 200 nM MitoQ and 50 μM RESV than other concentrations used. In another experiment, sperm pellet from thirty-two ejaculates was supplemented with 200 nM MitoQ and 50 μM RESV in the extender. Pre-freeze and post-thaw progressive motility and livability were higher (P < 0.05) in MitoQ (200 nM) than RESV (50 μM) treatment. MitoQ supplementation improved post-thaw membrane integrity (CFDA-PI) higher (P < 0.05) than RESV, however, hypo-osmotic swelling response observed no improvement with RESV treatment. Post-thaw LPO rate was lower (P < 0.05) and Bovine cervical mucus penetration was higher (P < 0.05) in MitoQ than RESV treatment. In post-thaw semen, MitoQ showed higher (P < 0.05) proportion of acrosome intact (FITC-PNA), live non-apoptotic (P < 0.01) sperm with a higher reduction (P < 0.05) in membrane scrambling. MitoQ improved (P < 0.01) proportion of sperm with high Mitochondrial Membrane Potential and low LPO (P < 0.01) than RESV treatment. In conclusion, improvement in post-thaw in-vitro sperm functions and cryo-tolerance was more evident in MitoQ than RESV supplemented buffalo bull semen. Our study provides a better strategy to mitigate oxidative stress by enhancing mitochondrial antioxidant system with targeted antioxidants than cytosolic antioxidant supplementation.
本研究比较了靶向线粒体（Mitoquinone，MitoQ）和非靶向胞质抗氧化剂（Resveratrol，RESV）对冷冻水牛公牛精液脂质过氧化（LPO）和体外精子功能的影响。为了优化添加剂的浓度，从 24 个精液样本中获得的精子沉淀分别用不同浓度的 MitoQ（20 nM、100 nM、200 nM）和 RESV（10 μM、25 μM、50 μM）在扩展剂中与对照组进行补充。与其他使用浓度相比，200 nM MitoQ 和 50 μM RESV 处理后的解冻后精子活力、存活率和膜完整性更高（P<0.05）。在另一个实验中，从 32 个精液样本中获得的精子沉淀在扩展剂中添加 200 nM MitoQ 和 50 μM RESV。预冷冻和解冻后，运动能力和存活率较高（P<0.05）MitoQ（200 nM）处理组高于 RESV（50 μM）处理组。与 RESV 处理相比，MitoQ 处理可提高解冻后膜完整性（CFDA-PI）（P<0.05），但 RESV 处理后低渗肿胀反应未见改善。解冻后 LPO 率较低（P<0.05），牛宫颈黏液穿透率较高（P<0.05）MitoQ 处理组高于 RESV 处理组。在解冻后的精液中，MitoQ 处理组显示出更高的（P<0.05）顶体完整（FITC-PNA）比例、活非凋亡（P<0.01）精子比例，并降低了（P<0.05）膜重排。与 RESV 处理相比，MitoQ 处理可提高（P<0.01）具有高线粒体膜电位和低 LPO（P<0.01）的精子比例。总之，与 RESV 处理相比，MitoQ 处理可明显改善解冻后体外精子功能和耐冷冻性。我们的研究提供了一种更好的策略，通过增强靶向抗氧化剂的线粒体抗氧化系统来减轻氧化应激，而不是通过胞质抗氧化剂补充。
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