CN108849860A - 一种抗氧化应激的精液冷冻保护剂 - Google Patents

一种抗氧化应激的精液冷冻保护剂 Download PDF

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Publication number
CN108849860A
CN108849860A CN201810871030.8A CN201810871030A CN108849860A CN 108849860 A CN108849860 A CN 108849860A CN 201810871030 A CN201810871030 A CN 201810871030A CN 108849860 A CN108849860 A CN 108849860A
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sperm
semen
added
freezing
cryoprotectant
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王红燕
马良宏
颜贝
王燕蓉
裴秀英
周岳
裴承斌
田稼
徐焱焱
王红红
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Ningxia Medical University
General Hospital of Ningxia Medical University
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Ningxia Medical University
General Hospital of Ningxia Medical University
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0205—Chemical aspects
    • A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N1/00—Preservation of bodies of humans or animals, or parts thereof
    • A01N1/02—Preservation of living parts
    • A01N1/0205—Chemical aspects
    • A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
    • A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Dentistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Environmental Sciences (AREA)
  • Biophysics (AREA)
  • Physiology (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

本发明公开了一种抗氧化应激的精液冷冻保护剂，该精液冷冻保护剂内添加有400ug/ml的枸杞多糖，通过以下方法制备所得：称取40mg枸杞多糖,1.5g葡萄糖，1.72g甘氨酸，1.72g枸橼酸钠，用52ml超纯水溶解后，加入28ml的甘油，滤过，加入20ml的蛋黄，拌匀，即得100ml抗氧化应激的精液冷冻保护剂。本发明的精液冷冻保护剂可增强冻后精子的前向运动百分率、复苏率、线粒体膜电位正常百分率、顶体反应正常百分率，降低氧化应激性DNA损伤(核碎裂指数DFI)、冻后精子ROS和丙二醛降低从而提高了精液的抗氧化能力，改善了冻后精液的质量。

Description

一种抗氧化应激的精液冷冻保护剂
技术领域
本发明涉及生殖医学领域，具体涉及一种抗氧化应激的精液冷冻保护剂。
背景技术
目前人类精液冷冻保存技术已经相对成熟并在临床得到广泛应用，成为辅助生殖技术(ART)领域中的重要技术。研究表明，精液冷冻过程中，冷冻刺激导致过量活性氧(ROS)的产生，从而使精子处于氧化应激状态，导致冷冻复苏后引起精子氧化应激性DNA损伤、精子活力和精子受精等精子功能的损伤，所以冷冻可以明显通过降低精子的抗氧化能力影响精子的功能，如何改进精液保护剂，降低复苏后精子DNA损伤，提高精子活力成为精液冷冻技术的发展趋势。
因为精液冷冻技术不可避免的氧化应激等体外干扰因素可能通过表观遗传修饰影响配子内印记基因的表达，导致配子的表观遗传信息改变。目前众多的试管婴儿来自于冻精或冻胚后移植，随着人们对生殖质量尤其遗传问题的日愈重视，评估精液冷冻技术的表观遗传安全性，改良冷冻保护剂，降低保护剂毒性提高其遗传安全性，改善妊娠结局势在必行。
研究已证实，精液冷冻保护剂加入超氧化物歧化酶(SOD)、维生素E等抗氧化剂，通过清除氧自由基，抵抗各种氧化因子对生物组织的氧化性破坏，减少冷冻对精液刺激产生的过量ROS，保护精子线粒体功能和精子生物膜结构的完整性，从而改善冷冻复苏后精子质量。
发明内容
为解决上述问题，本发明提供了一种抗氧化应激的精液冷冻保护剂，可以提高精液的抗氧化效应，降低冷冻刺激过程中的精子的氧化应激反应。
为实现上述目的，本发明采取的技术方案为：
一种抗氧化应激的精液冷冻保护剂，该精液冷冻保护剂内添加有
400ug/ml的枸杞多糖，通过以下方法制备所得：
称取40mg枸杞多糖,1.5g葡萄糖，1.72g甘氨酸，1.72g枸橼酸钠，用52ml超纯水溶解后，加入28ml的甘油，滤过，加入20ml的蛋黄，拌匀，即得100ml抗氧化应激的精液冷冻保护剂。
本发明具有以下有益效果：
本发明的精液冷冻保护剂可增强冻后精子的前向运动百分率、复苏率、线粒体膜电位正常百分率、顶体反应正常百分率，降低氧化应激性DNA损伤(核碎裂指数DFI)、冻后精子ROS和丙二醛降低从而提高了精液的抗氧化能力，改善了冻后精液的质量。
具体实施方式
为了使本发明的目的及优点更加清楚明白，以下结合实施例对本发明进行进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。
本发明实施例的一种抗氧化应激的精液冷冻保护剂以超纯水为基液，枸橼酸钠、甘氨酸和葡萄糖为缓冲液，甘油和蛋黄作为渗透性和非渗透性保护剂成分，添加具有抗氧化和对生殖细胞具有保护性作用的枸杞多糖形成，通过以下方法制备所得：
称取40mg枸杞多糖,1.5g葡萄糖，1.72g甘氨酸，1.72g枸橼酸钠，用52ml超纯水溶解后，加入28ml的甘油，滤过，加入20ml的蛋黄，拌匀，即得100ml抗氧化应激的精液冷冻保护剂。
本具体实施通过改良保护剂，加入抗氧化剂枸杞多糖，通过清除氧自由基，抵抗各种氧化因子对生物组织的氧化性破坏，降低冷冻刺激过程中的精子的氧化应激反应；保护精子线粒体功能和精子生物膜结构的完整性；经验证，其可增强冻后精子的前向运动百分率、复苏率、线粒体膜电位正常百分率、顶体反应正常百分率，降低氧化应激性DNA损伤(核碎裂指数DFI)、冻后精子的凋亡率和过量活性氧(ROS)和丙二醛含量降低从而提高了精液的抗氧化能力，改善了冻后精液的质量。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以作出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (2)

1.一种抗氧化应激的精液冷冻保护剂，其特征在于：该精液冷冻保护剂内添加有400ug/ml的枸杞多糖。
2.如权利要求1所述的一种抗氧化应激的精液冷冻保护剂，其特征在于：所述精液冷冻保护剂通过以下方法制备所得：
称取40mg枸杞多糖,1.5g葡萄糖，1.72g甘氨酸，1.72g枸橼酸钠，用52ml超纯水溶解后，加入28ml的甘油，滤过，加入20ml的蛋黄，拌匀，即得100ml抗氧化应激的精液冷冻保护剂。
CN201810871030.8A 2018-08-02 2018-08-02 一种抗氧化应激的精液冷冻保护剂 Pending CN108849860A (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112931483A (zh) * 2021-01-29 2021-06-11 海南晨海水产有限公司 一种保持高存活的浪人鲹精子冷冻液及其制备方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103262837A (zh) * 2013-05-20 2013-08-28 北京市农林科学院 一种家畜精液冷冻保护剂及其应用
CN107535481A (zh) * 2017-09-29 2018-01-05 界首市国启家庭农场 一种猪精液冷冻保护剂

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103262837A (zh) * 2013-05-20 2013-08-28 北京市农林科学院 一种家畜精液冷冻保护剂及其应用
CN107535481A (zh) * 2017-09-29 2018-01-05 界首市国启家庭农场 一种猪精液冷冻保护剂

Non-Patent Citations (2)

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何涛 等: "枸杞多糖对猪精液冷冻效果影响的研究", 《陕西农业科学》 *
李方舟 等: "海带多糖和枸杞多糖对山羊精液冷冻保存效果的影响", 《家畜生态学报》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112931483A (zh) * 2021-01-29 2021-06-11 海南晨海水产有限公司 一种保持高存活的浪人鲹精子冷冻液及其制备方法
CN112931483B (zh) * 2021-01-29 2022-05-24 海南晨海水产有限公司 一种保持高存活的浪人鲹精子冷冻液及其制备方法

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