Ahmed Hussain, Jahan Sarwat, Ijaz Muhammad Umar, Riaz Mehreen, Ullah Farhad, Saqib Najm Us
Department of Zoology, University of Buner, Khyber Pakhtunkhwa, KP, Pakistan; Reproductive Physiology Laboratory, Department of Animal Sciences, Quaid-i-Azam University QAU, Islamabad, Pakistan.
Reproductive Physiology Laboratory, Department of Animal Sciences, Quaid-i-Azam University QAU, Islamabad, Pakistan.
Cryobiology. 2022 Aug;107:42-47. doi: 10.1016/j.cryobiol.2022.05.004. Epub 2022 May 26.
Current study was conducted to appraise the cryoprotective influence of crocetin on quality, oxidative status, and fertility potential of bubaline spermatozoa. Collected semen from four bulls was diluted in five aliquots with (10 μM, 5 μM, 2 μM, 1 μM, and control [0 μM] supplementation of crocetin). After gentle dilution (37 °C), cooling (4 °C, in 2 h), equilibration (4 °C, for 4 h) and packaging of samples was done in straws (polyvinyl French, 0.5 ml), and then frozen (programmable cell freezer). This study established that crocetin supplementation significantly (p < 0.05) improves CASA (Computer Assisted Sperm motion Analyzer) total motility (%), rapid velocity (%), average-path, and curved-line velocities (μm/sec, 10 μM vs. control), and progressive motility (%), straight-line velocity (μm/sec), total antioxidant capacity (TAC, μMol/l), ATP concentrations (nmol/million), and fertility potential (%) (10 μM vs. control, and 1 μM), and mitochondrial potential (%) of buffalo spermatozoa (5, and 10 μM vs. control). Crocetin supplementation significantly (p < 0.05) alleviates DNA fragmentation, seminal plasma ROS (10 RLU/20/25 million, RLU = Relative light unit) levels, and lipid peroxidation (LPO, μMol/ml) in buffalo spermatozoa (10 μM vs. control). In a nutshell, crocetin supplementation improves post-thaw quality by means of motility parameters, motion kinematics, TAC, and ATP concentrations, and fertility potential, and abolished DNA fragmentation parameters, seminal plasma ROS, and LPO concentrations of buffalo spermatozoa. The exact mechanism by which crocetin acts are not fully elucidated; however, it is probable to speculate that the reduction in ROS, and LPO recorded in this study may be related to scavenging ability of this antioxidant during cryopreservation.
本研究旨在评估藏红花素对水牛精子质量、氧化状态和生育潜力的冷冻保护作用。从四头公牛采集的精液用藏红花素（10 μM、5 μM、2 μM、1 μM和对照[0 μM]）分成五等份进行稀释。轻轻稀释（37°C）后，冷却（4°C，2小时），平衡（4°C，4小时），然后将样品包装在吸管（聚乙烯法式，0.5毫升）中，接着进行冷冻（程序降温细胞冻存仪）。本研究表明，添加藏红花素显著（p < 0.05）提高了计算机辅助精子运动分析系统（CASA）的总活力（%）、快速运动速度（%）、平均路径速度和曲线速度（μm/秒，10 μM与对照相比），以及渐进性活力（%）、直线速度（μm/秒）、总抗氧化能力（TAC，μMol/l）、ATP浓度（nmol/百万）和生育潜力（%）（10 μM与对照相比，以及1 μM），还有水牛精子的线粒体膜电位（%）（5和10 μM与对照相比）。添加藏红花素显著（p < 0.05）减轻了水牛精子的DNA片段化、精浆活性氧（10 RLU/20/25百万，RLU = 相对光单位）水平和脂质过氧化（LPO，μMol/ml）（10 μM与对照相比）。简而言之，添加藏红花素通过运动参数、运动学、TAC和ATP浓度以及生育潜力提高了解冻后的质量，并消除了水牛精子的DNA片段化参数、精浆活性氧和LPO浓度。藏红花素发挥作用的确切机制尚未完全阐明；然而，可以推测本研究中记录的活性氧和LPO的减少可能与这种抗氧化剂在冷冻保存期间的清除能力有关。
