用丁基羟基甲苯和胆固醇负载的甲基-β-环糊精进行预冷冻处理可提高冷冻保存的公猪精液质量。

Pre-freezing Treatment with Butylated Hydroxytoluene and Cholesterol-loaded Methyl-β-cyclodextrin Improves Quality of Cryopreserved Boar Semen.

作者信息

Baishya S K, Biswas R K, Govindasamy K, Deka B C, Sinha S, Singh M

机构信息

Krishi Vigyan Kendra, ICAR Research Complex for NEH Region Nagaland Centre, Longsachung, Nagaland.

Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary Science, Assam Agricultural University, Khanapara, Assam.

出版信息

Cryo Letters. 2018 Sep/Oct;39(5):336-344.

PMID:30963166

Abstract

BACKGROUND

Pre-freezing treatment of boar sperm with additives improves the quality of post-thaw sperms.

OBJECTIVES

The study aimed to determine the efficacy of butylated hydroxy-toluene (BHT) and cholesterol-loaded methyl-β-cyclodextrin (CLC) for the improvement of the frozen-thawed boar sperm quality.

METHODS

Split samples of 30 ejaculates from six boars were cryopreserved in lactose-egg yolk-glycerol extender containing BHT (0.2 mM), CLC (5 mg/ 200-240 x 10 sperm) or BHT (0.2 mM) plus CLC (5 mg per 200-240 x 10 sperm). Semen samples were evaluated for motility, membrane integrity, acrosomal status, mitochondrial membrane potential (MMP), DNA integrity and lipid peroxidation after equilibration and after freezing.

RESULTS

The addition of BHT and CLC into the extender significantly improved (P<0.05) plasma membrane integrity and decreased (P<0.05) lipid peroxidation after freezing. Post-thaw motility and live intact acrosome were significantly (P<0.05) higher in the extenders with BHT or BHT plus CLC. The post-thaw MMP of viable spermatozoa and DNA integrity were not affected. BHT plus CLC showed a significant (P<0.05) improvement on motility as compared to BHT and CLC alone.

CONCLUSION

Treatment of boar spermatozoa with BHT and CLC improved post-thaw sperm quality.

摘要

背景

用添加剂对公猪精子进行预冷冻处理可提高解冻后精子的质量。

目的

本研究旨在确定丁基羟基甲苯（BHT）和胆固醇负载的甲基-β-环糊精（CLC）对改善冷冻解冻公猪精子质量的效果。

方法

将来自6头公猪的30份射精样本分成几份，保存在含有BHT（0.2 mM）、CLC（5 mg/200 - 240×10精子）或BHT（0.2 mM）加CLC（每200 - 240×10精子5 mg）的乳糖-蛋黄-甘油稀释液中。在平衡后和冷冻后，对精液样本进行活力、膜完整性、顶体状态、线粒体膜电位（MMP）、DNA完整性和脂质过氧化评估。

结果

在稀释液中添加BHT和CLC可显著提高（P<0.05）冷冻后的质膜完整性，并降低（P<0.05）脂质过氧化。在含有BHT或BHT加CLC的稀释液中，解冻后的活力和完整顶体活精子显著更高（P<0.05）。解冻后活精子的MMP和DNA完整性不受影响。与单独使用BHT和CLC相比，BHT加CLC在活力方面有显著（P<0.05）改善。