抗冷冻蛋白Ⅲ减少家兔卵巢组织冷冻损伤的效果观察  

Effect of Antifreeze Protein Ⅲ on Reducing Freeze Damage of Rabbit Ovarian Tissue in Cryopreservation

作　　者：曾琴[1] 刘伟信[1,2] 罗孟军[3] 王凯[4] 徐红[1] 

机构地区：[1]成都中医药大学第二临床医学院,四川成都610075 [2]四川省妇幼保健院,四川成都610045 [3]成都市计划生育指导所,四川成都610031 [4]川北医学院,四川南充637000

出　　处：《实用妇产科杂志》2016年第5期383-387,共5页Journal of Practical Obstetrics and Gynecology

基　　金：国家自然科学基金项目(编号:81072834)

摘　　要：目的:评价抗冷冻蛋白Ⅲ(AFPⅢ)对玻璃化冷冻家兔卵巢组织的影响。方法:收集家兔卵巢30只,随机分为新鲜卵巢组、添加AFPⅢ玻璃化冷冻组(AFPⅢ终浓度为500 ng/ml)和常规玻璃化冷冻组,各组10只,解冻后分析各组卵巢的组织学结构、卵泡形态正常率、卵巢组织超微结构、卵母细胞凋亡率及卵泡存活率。结果:新鲜卵巢组的卵泡形态正常率(91.6%)、卵泡存活率(81.75%)显著高于两冷冻组(P<0.01),卵母细胞凋亡率(12.0%)显著低于两冷冻组(P<0.01);添加AFPⅢ玻璃化冷冻组卵泡形态正常率(77.5%)、卵泡存活率(45.31%)显著高于常规玻璃化冷冻组(分别为62.1%、37.25%)(P<0.01),卵母细胞凋亡率(25.8%)显著低于常规玻璃化冷冻组(41.2%)(P<0.01)。结论:家兔冷冻卵巢组织的卵泡形态正常率、卵泡存活率显著低于新鲜组织,冷冻保护剂中加入AFPⅢ可减少家兔卵巢组织冷冻损伤。Objective：To observe the effects of antifreeze protein III （AFPIII） on rabbit ovarian tissue cryopr- eservation. Methods：30 ovaries were randomly divided into three groups： fresh control group, AFP III treatment group（AFPIII 500 ng/ml） and vitrification control group. Ovarian tissue ultrastrcture,the rate of normal morpho- logical follicles, oocyte apoptosis rate, and the live follicles rate were evaluated by using electron microscope, his- tology,TUNEL assays, and Live/Dead-ViabUity/Cytotoxicity Kit. Results： The normal morphological follicles rate and the live follicles rate of fresh ovarian tissue group were higher than that of two vitrified-warmed ovarian tissue groups（ P 〈0. 01 ）, and these rates of AFP III treatment group were significantly higher than that of vitrification control group（ P〈0. 01 ）. The oocyte apoptosis rate of the fresh control group was significantly lower than that of the other two vitrified-warmed ovarian tissue groups（ P 〈 0. 01 ）, moreover the AFP III treatment group was signifi- cantly lower than that of the vitrification control group（ P 〈0. 01 ）. Conclusions.The follicle survival rate of two vit- rified-warmed ovarian tissue groups was lower than that of the fresh ovarian tissue group, but supplementation of AFPIII in vitrification medium can significantly improve the follicle viability of cryopreserved rabbit ovarian tissue, which provided theoretical basis for the application of AFPIII to ovarian tissue cryopreservation.

关 键 词：抗冷冻蛋白Ⅲ 卵巢组织 玻璃化冷冻 家兔 

分 类 号：R321-33[医药卫生—人体解剖和组织胚胎学]