精子冷冻保存可能会导致精子DNA受损,膜,和冻融过程引起的整体运动。白细胞介素-6(IL-6)是一种在生殖过程中具有多种作用的多功能细胞因子。然而,补充IL-6对冷冻保存的公羊精子的影响尚未得到彻底研究.因此,本研究旨在评估IL-6对冷冻保存的公羊精子质量的影响。收集了公羊精液,池化,并用补充有0、50、100和200ng/mLIL-6的tris-柠檬酸大豆卵磷脂补充剂进行扩展。样本经历了标准的冷冻方案,精子质量,运动学参数,超微结构,并对冷冻保存的公羊精子进行了分子对接评估。结果表明,精子运动学,生存能力,渐进运动,通过添加100或200ng的IL-6/mL(p<0.05),膜的完整性显着增强。补充有100或200ng/mLIL-6的精液也表现出更高的精子运动学百分比,包括DAP,DCL,DSL,VSL,VAP,VCL,ALH,与其他组相比(p<0.05)。IL-6补充增强顶体完整性,与未处理组相比,解冻后的公羊精子中的caspase-3活性降低(p<0.05)。补充IL-6(200ng/mL)显着降低氧化生物标志物(NO,MDA,和H2O2)(p<0.001),并提高了总抗氧化能力(p<0.05)。精子损伤的百分比(尾巴,头部,和中片)通过补充IL-6显着降低(p<0.05)。电子显微照片显示,补充100或200ng/mLIL-6保护顶体稳定性,质膜完整性,并维持了冷冻保存的公羊精子的超微结构完整性。对接探索表明与精子功能生物标志物的结合亲和力更高,包括caspase3,BCL2和PSMA6,结合能为-52.30kcal/mol,-56.04kcal/mol,和-57.06千卡/摩尔,分别。总之,在冷冻延长剂中添加IL-6可以提高冷冻保存的公羊精子的解冻后质量。
Sperm cryopreservation can lead to subfertility due to potential damage to sperm DNA, membranes, and overall motility caused by the freeze-thaw process. Interleukin-6 (IL-6) is a versatile cytokine with various roles in reproductive processes. However, the impacts of IL-6 supplementation on cryopreserved
ram sperm have not been thoroughly investigated. Therefore, this study aims to assess the influence of IL-6 on the sperm quality of cryopreserved
ram sperm.
Ram semen was collected, pooled, and extended with tris-citrate soybean lecithin extender supplemented with 0, 50, 100, and 200 ng/mL of IL-6. The samples experienced a standard freezing protocol, and sperm quality, kinematic parameters, ultrastructure, and molecular docking of cryopreserved
ram spermatozoa were evaluated. The results showed that sperm kinematics, viability, progressive motility, and membrane integrity were significantly enhanced by the addition of 100 or 200 ng of IL-6/mL (p < 0.05). Semen supplemented with 100 or 200 ng/mL of IL-6 also exhibited higher percentages of sperm kinematics, including DAP, DCL, DSL, VSL, VAP, VCL, and ALH, compared to other groups (p < 0.05). IL-6 supplementation enhanced acrosome integrity, and reduced caspase-3 activity in post-thawed
ram spermatozoa (p < 0.05) compared to untreated group. Supplementation with IL-6 (200 ng/mL) significantly decreased oxidative biomarkers (NO, MDA, and H2O2) (p < 0.001) and improved total antioxidant capacity (p < 0.05). The percentage of sperm damage (tail, head, and midpiece) was significantly reduced by IL-6 supplementation (p < 0.05). Electron micrographs showed that supplementation with 100 or 200 ng/mL IL-6 protected acrosome stability, plasma membrane integrity, and sustained the ultrastructure integrity of cryopreserved ram spermatozoa. The docking exploration indicates a higher binding affinity with sperm function biomarkers, including caspase 3, BCL2, and PSMA6, with binding energies of - 52.30 kcal/mol, - 56.04 kcal/mol, and - 57.06 kcal/mol, respectively. In conclusion, the addition of IL-6 to the freezing extender can enhance the post-thaw quality of cryopreserved ram spermatozoa.