Kumar Pradeep, Kumar Dharmendra, Sikka P, Singh P
ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana, India.
ICAR-Central Institute for Research on Buffaloes, Hisar, Haryana, India.
Anim Reprod Sci. 2015 Jan;152:26-31. doi: 10.1016/j.anireprosci.2014.11.015. Epub 2014 Dec 2.
The variety of mammalian cells has been successfully cryopreserved by use of the silk protein sericin due to its strong free-radical-scavenging and potent antioxidant activity. The present study was conducted to examine the protective role of sericin on buffalo spermatozoa during cryopreservation. Semen of four breeding bulls was collected twice a week using artificial vagina technique. The ejaculates of four bulls were pooled, divided into five equal fractions, diluted with the extender supplemented with different concentrations of sericin (0, 0.25, 0.5, 1.5 and 2%) and then cryopreserved. Post-thawed motility was objectively assessed by computer assisted sperm analyzer. Sperm plasma membrane integrity was assessed by hypo-osmotic swelling test (HOST). Malondialdehyde (MDA) concentration, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were determined in frozen-thawed extended seminal plasma by spectrophotometry. The extender supplemented with 0.25, 0.5 and 1% sericin resulted in the higher sperm motility and GPx acivity. Furthermore, plasma membrane integrity and SOD activity were found to be higher (P<0.05) in group supplemented with 0.25 and 0.5% sericin (P<0.05). The MDA concentration was found to be significantly lower (P<0.05) in 0.25 and 0.5% sericin treated groups than control and other treated groups. In conclusion, the supplementation of 0.25-0.5% sericin in semen extender improves frozen-thawed semen quality through protecting sperm from oxidative stress.
由于丝蛋白丝胶具有强大的自由基清除能力和有效的抗氧化活性,多种哺乳动物细胞已成功通过使用丝胶进行冷冻保存。本研究旨在探讨丝胶在水牛精子冷冻保存过程中的保护作用。使用人工阴道技术,每周两次采集四头种公牛的精液。将四头公牛的射精混合,分成五个相等的部分,用添加不同浓度丝胶(0、0.25、0.5、1.5和2%)的稀释液稀释,然后进行冷冻保存。解冻后的活力通过计算机辅助精子分析仪进行客观评估。精子质膜完整性通过低渗肿胀试验(HOST)进行评估。通过分光光度法测定冻融后精液稀释液中的丙二醛(MDA)浓度、谷胱甘肽过氧化物酶(GPx)和超氧化物歧化酶(SOD)活性。添加0.25、0.5和1%丝胶的稀释液可使精子活力和GPx活性更高。此外,添加0.25和0.5%丝胶的组中质膜完整性和SOD活性更高(P<0.05)。在0.25和0.5%丝胶处理组中,MDA浓度显著低于对照组和其他处理组(P<0.05)。总之,在精液稀释液中添加0.25 - 0.5%的丝胶可通过保护精子免受氧化应激来提高冻融精液质量。