Abstract：
Semen cryopreservation causes extensive chemical and physical damage to sperm structure, which generates premature aging and reduces viability and fertility of spermatozoa. The addition of antioxidants to freezing extenders can reduce the oxidative damage caused by excessive generation of reactive oxygen species (ROS), and the premature aging could be reduced by adding an enzyme inhibitor that prevents an anticipated capacitation. The aim of this study was to evaluate the in vitro effect of quercetin (Q), L-ergothioneine (E) and H89 addition to cryopreserved equine spermatozoa. Six experimental groups were stablished: control, Q, E, H89, H89Q and H89E. The analyzed parameters were sperm motility and kinematic using computer assisted sperm analysis (CASA), plasma membrane functionality with the hypoosmotic swelling test (HOST) and fertilizing capability with in vitro heterologous fertilization. Quercetin reduced curvilinear velocity (VCL) and increased beat-cross frequency (BCF), while its combination with H89 (H89Q) reduced total motility, progressive motility, VCL and hyperactive sperm (HA). Likewise, H89 and its combination with E (H89E) decreased VCL and amplitude of lateral head displacement (ALH). No significant differences were observed among treatments for membrane functionality and fertilizing capacity of sperm. In conclusion H89 in combination with Q and E reduced sperm motility or some kinematic parameters. However, they did not influence plasma membrane functionality and in vitro fertilizing capacity of frozen-thawed equine semen.
摘要：
精液冷冻保存会对精子结构造成广泛的化学和物理损伤，这会导致精子过早衰老并降低精子的生存能力和生育能力。在冷冻填充剂中添加抗氧化剂可以减少由活性氧（ROS）过量产生引起的氧化损伤，并且通过添加防止预期获能的酶抑制剂可以减少过早老化。本研究的目的是评估槲皮素（Q）的体外作用，L-麦角硫因（E）和H89添加到冷冻保存的马精子中。建立了六个实验组：对照组，Q,E,H89、H89Q和H89E。使用计算机辅助精子分析（CASA）分析的参数为精子运动性和运动学，具有低渗肿胀试验（HOST）的质膜功能和体外异源受精的受精能力。槲皮素降低曲线速度（VCL）和增加拍频（BCF），虽然它与H89（H89Q）的组合降低了总运动性，渐进运动，VCL和多动精子（HA）。同样，H89及其与E（H89E）的组合降低了VCL和头部侧向位移（ALH）的幅度。在膜功能和精子受精能力的处理之间没有观察到显着差异。总而言之，H89与Q和E结合会降低精子运动能力或某些运动学参数。然而,它们不影响冻融马精液的质膜功能和体外受精能力。
