卵巢组织移植, 卵巢组织冷冻, 卵泡计数, 细胞凋亡, 血管内皮生长因子," /> 卵巢组织移植, 卵巢组织冷冻, 卵泡计数, 细胞凋亡, 血管内皮生长因子,"/> Ovarian tissue transplantation, Ovarian tissne cryofireservation, Follicle count , Cell apoptosis , Vascular endothelial growth factor,"/> <span style="line-height:2;font-size:14px;">大鼠卵巢组织玻璃化冷冻及自体移植</span><span style="line-height:2;font-size:14px;">对卵泡的影响</span>
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发育医学电子杂志  2019, Vol. 7 Issue (2): 130-135    DOI: 10.3969/j.issn.2095-5340.2019.02.011
  生殖胚胎   论著 |
大鼠卵巢组织玻璃化冷冻及自体移植对卵泡的影响
邢彦彦  赵玮  李文
1. 海军军医大学附属第二医院上海长征医院 生殖中心,上海 200433;2. 复旦大学附属金山医院 妇产科,上海 200540
Effects of follicle for vitrification and autotransplantation on loss of ovarian tissue in rats
XING Yan-yan  ZHAO Wei  LI Wen
1.Reproductive Medicine Center , Shanghai Changzheng Hospital, the Second Military Medical University, Shanghai 200433, China; 2.Department of Gynaecology and Obstetrics, Jinshan Hospital, Fudan University, Shanghai 200540, China
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摘要 【摘要】 目的 探讨大鼠卵巢组织冷冻及自体移植对卵泡形态和数量的影响。 方法 8 周龄 SD 雌性
大鼠随机分为对照组、移植组、冷冻组,每组 3 只,6 个卵巢组织。麻醉后切除双侧卵巢组织。对照组
直接固定并进行 HE 和免疫组化染色。移植组进行肾被膜下自体移植,4 周后取移植卵巢组织。冷冻
组采用玻璃化冷冻方法,4 周后取出复苏。观察各组卵巢组织形态学,并对卵泡计数。采用 TUNEL 荧光检测卵巢细胞凋亡水平。采用免疫组织化学法检测各组 VEGF 表达情况。统计学方法采用单因素方差分析和独立样本 t 检验。 结果 对照组、冷冻组、移植组每张切片的卵泡均数分别为(32.3±4.2)、(27.2±7.0)、(22.5±1.5)个,冷冻组及移植组均低于对照组,移植组低于冷冻组,差异有统计学意义(F=41.746,P<0.05)。对照组、冷冻组、移植组的细胞凋亡指数分别为(2.8±0.3)、(5.5±1.0)与(14.2±2.8)(F=60.788,P<0.05),冷冻组及移植组的凋亡指数均高于对照组,移植组高于冷冻组,差异有统计学意义(P<0.05)。对照组、冷冻组和移植组免疫组织化学染色 VEGF 阳性细胞百分比分别为(79.5±5.3)%、(63.2±6.5)% 与(48.5±2.8)%(F=45.871,P<0.05),冷冻组和移植组的 VEGF 表达低于对照组,移植组VEGF 表达低于冷冻组,差异均有统计学意义(P<0.05)。 结论 卵巢组织冷冻或自体移植过程中均伴有卵泡丢失、卵巢组织 VEGF 表达下降。提高移植卵巢组织的 VEGF 表达,可能对促进移植物血管形成、提高卵巢移植效率有积极作用。
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邢彦彦 赵玮 李文
关键词:  卵巢组织移植')" href="#">卵巢组织移植  卵巢组织冷冻  卵泡计数  细胞凋亡  血管内皮生长因子    
Abstract: 【Abstract】 Objective To study the effect of follicular morphology and amount for cyryopreservation
and autotransplantation on ovarian tissue in rats. Methods Female SD rats aged 8 weeks were randomly divided into 3 groups: control group, autotransplantation group and vitrification group, each group had 3 SD rats and 6 ovarian tissues. The bilateral ovarian tissues of cases were removed after anesthesia. HE and immunohistochemical staining were performed and fixed directly for control group. The autotransplantation under renal capsule was underwent and ovarian tissue was taken after 4 weeks in autotransplantation group. Vitrification was used in the vitrification group and that was taken and resuscitated after 4 weeks. Ovarian histomorphology was observed for all of the groups and the follicles were counted. TUNEL ?uorescence assay was used to detect the level of apoptotic cells in ovarian tissue and VEGF expression in each groupwas detected by immunohistochemistry. The statistical methods were carried out by single factor analysis of variance and independent sample t test. Results The follicle counts of control group, vitrification group and transplantation group were (32.3±4.2), (27.2±7.0) and (22.5±1.5) respectively. The vitrification group and transplantation group were lower than that in the control group, and the transplantation group was lower than that in the vitrification group with statistical significance (F=41.746, P<0.05). The cell apoptotic indexs of control group, vitrification group and transplantation group were (2.8±0.3), (5.5±1.0) and (14.2±2.8) respectively (F=60.788,P<0.05). The vitrification group and transplantation group were higher than the control group in cell apoptotic index, and the transplantation group was higher than that in the vitrification group with statistical significance (P<0.05). The percentages of positive cells for VEGF immunohistochemical staining in control group, vitrification group and transplantation group were (79.5±5.3)%, (63.2±6.5)% and (48.5±2.8)% respectively (F=45.871, P<0.05). The VEGF expression of vitrification group and transplantation group was lower than that in the control group, and the transplantation group was lower than that in the vitrification group, there were statistical significance (P<0.05).  Conclusions  Ovarian tissue cryopreservation or ovarian autotransplantation are all accompanied by follicular loss and decrease of VEGF expression. Increasing VEGF expression of ovarian tissue has positive effect for promoting graft angiogenesis and improving the efficiency of ovarian transplantation.
Key words:  Ovarian tissue transplantation')" href="#">Ovarian tissue transplantation    Ovarian tissne cryofireservation    Follicle count    apoptosis ')" href="#"> Cell apoptosis    Vascular endothelial growth factor
收稿日期:  2019-02-13                出版日期:  2019-04-30      发布日期:  2019-05-09      期的出版日期:  2019-04-30
基金资助: 军事医学创新工程(16JS007);上海市科委医学引导项目(16411963500);上海市科委标准项目
(16DZ0500402)
通讯作者:  李文    E-mail:  773791143@qq.com
引用本文:    
邢彦彦 赵玮 李文. 大鼠卵巢组织玻璃化冷冻及自体移植对卵泡的影响[J]. 发育医学电子杂志, 2019, 7(2): 130-135.
XING Yan-yan ZHAO Wei LI Wen. Effects of follicle for vitrification and autotransplantation on loss of ovarian tissue in rats. Journal of Developmental Medicine(Electronic Version), 2019, 7(2): 130-135.
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http://www.fyyxzz.com/CN/10.3969/j.issn.2095-5340.2019.02.011  或          http://www.fyyxzz.com/CN/Y2019/V7/I2/130
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