William R. Pritchard Veterinary Medical Teaching Hospital, School of Veterinary Medicine, University of California, 1 Garrod Drive, Davis, CA, 95616, USA.
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, 1 Garrod Drive, Davis, CA, 95616, USA.
Theriogenology. 2019 Aug;134:83-89. doi: 10.1016/j.theriogenology.2019.05.009. Epub 2019 May 16.
In equids, it is common to inject lidocaine into the testicles at the time of routine castration to provide analgesia. The effects of lidocaine on equine sperm have not been evaluated in vitro or on epididymal sperm collected following castration. The aims of this study were to determine effects of clinically relevant doses of lidocaine on equine spermatozoa in vitro using freshly collected semen and to compare the characteristics of epididymal spermatozoa after routine castration with or without intra-testicular lidocaine administration. We hypothesized that increasing concentrations of lidocaine would decrease total motility (TM), progressive motility (PM), velocity of the average path (VAP), velocity of the curved line (VCL), linearity (LIN), normal morphology (M) and membrane integrity (MI). We also hypothesized that injection of intra-testicular lidocaine would decrease TM, PM, VAP, VCL, LIN, M, and MI following routine castration, epididymal flushing and cryopreservation. In experiment 1, sperm was collected from four stallions and mixed with lidocaine at concentrations of 1 μg/ml, 10 μg/ml, 100 μg/ml, 1,000 μg/ml and 10,000 μg/ml. M and MI were compared to the control sample at 0 and 48 h. Motility parameters were analyzed at 0, 2, 4, 6, 24, and 48 h. In experiment 2, 12 stallions were castrated under general anesthesia. One testicle was removed without the use of intra-testicular lidocaine and the other testicle was removed 10 min after injection of 10 ml of 2% lidocaine. Results: In experiment 1, fresh sperm showed no significant difference (p < 0.05) compared to control at either 1 μg/ml or 10 μg/ml concentrations of lidocaine. There were significant decreases in PM, VAP, VCL, and LIN at concentrations of 100μg/ml-10,000 μg/ml and for TM at lidocaine concentrations of 1,000-10,000 μg/ml compared to control. Morphology did not change at any lidocaine concentration. Membrane integrity decreased significantly at 10,000 μg/ml lidocaine. In the second experiment 1.03 ± 0.42 μg/ml lidocaine was detected in the epididymal flush of stallions treated with lidocaine. There were no significant differences in any measured parameters between the control and the lidocaine treated testicles. Intra-testicular lidocaine injection at the time of castration did not affect any measured parameters after epididymal flush. Lidocaine concentrations higher than 100 μg/ml in-vitro resulted in decreased motility parameters of the spermatozoa independent of exposure time.
在马属动物中,在常规去势时向睾丸内注射利多卡因以提供镇痛是很常见的。利多卡因对马属动物精子的影响尚未在体外或去势后采集的附睾精子中进行评估。本研究的目的是使用新鲜采集的精液,确定临床相关剂量的利多卡因对体外马属精子的影响,并比较常规去势时是否向睾丸内注射利多卡因对精子的特征的影响。我们假设,随着利多卡因浓度的增加,总活力(TM)、前向运动(PM)、平均路径速度(VAP)、曲线速度(VCL)、直线性(LIN)、正常形态(M)和膜完整性(MI)会降低。我们还假设,在常规去势、附睾冲洗和冷冻保存后,向睾丸内注射利多卡因会降低 TM、PM、VAP、VCL、LIN、M 和 MI。在实验 1 中,从四匹种马中采集精子,并将其与浓度为 1μg/ml、10μg/ml、100μg/ml、1000μg/ml 和 10000μg/ml 的利多卡因混合。在 0 和 48 小时时,将 M 和 MI 与对照样本进行比较。在 0、2、4、6、24 和 48 小时时分析运动参数。在实验 2 中,12 匹种马在全身麻醉下被去势。一只睾丸在不使用睾丸内利多卡因的情况下被切除,另一只睾丸在注射 2%利多卡因 10ml 10 分钟后被切除。结果:在实验 1 中,新鲜精子与对照相比,在 1μg/ml 或 10μg/ml 利多卡因浓度下均无显著差异(p<0.05)。在 100μg/ml-10000μg/ml 浓度下,PM、VAP、VCL 和 LIN 显著下降,在 1000-10000μg/ml 利多卡因浓度下 TM 显著下降,与对照相比。在任何利多卡因浓度下,形态均未发生变化。在 10000μg/ml 利多卡因时,膜完整性显著下降。在第二个实验中,在接受利多卡因治疗的种马的附睾冲洗液中检测到 1.03±0.42μg/ml 的利多卡因。在对照和利多卡因处理的睾丸之间,没有任何测量参数的差异。在去势时向睾丸内注射利多卡因不会影响附睾冲洗后的任何测量参数。体外浓度高于 100μg/ml 的利多卡因会导致精子运动参数下降,而与暴露时间无关。