新生大鼠卵巢体外培养中NT3与LIF对始基卵泡发育的影响  

Effects of NT3 and LIF on Development of Primordial Follicles During In Vitro Culture of Neonatal Rat Ovary

作　　者：冯月枝[1] 邱绮[1] 杨冬梓[1] 张清学[1] 

机构地区：[1]中山大学孙逸仙纪念医院生殖医学中心,广东广州510120

出　　处：《中山大学学报（医学科学版）》2016年第4期535-542,共8页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金(S2011010004662)

摘　　要：【目的】研究NT3对早期卵泡存活、发育、激素合成的影响及其可能的作用机制,同时探讨NT3和LIF在早期卵泡启动发育方面是否存在协同或叠加效应。【方法】分离新生4d大鼠卵巢随机分配至新鲜组、基础培养组、NT3组、LIF组和NT3+LIF组,在37℃培养箱中培养14 d,经固定、包埋、切片后进行组织形态学、PCNA免疫组化、TUNEL凋亡分析,RTPCR检测KL m RNA表达及检测培养液中激素水平。【结果】相比新鲜组和基础培养组,NT3组、LIF组、NT3+LIF组中的始基卵泡比例下降,生长卵泡比例升高,同时PCNA蛋白表达显著增加;TUNEL分析显示NT3组、LIF组、NT3+LIF组的凋亡指数均显著降低;NT3、LIF组、NT3+LIF组卵巢中KL m RNA的表达量分别是对照组的(3.38±0.43)倍、(2.02±0.20)倍、(2.07±0.21)倍。【结论】NT3在早期卵泡存活、发育中有促进作用,上调KL基因表达可能是NT3的作用机制之一。NT3和LIF单独或相加均能促进卵泡发育,但两者之间无明显协同或叠加效应。[ Objective ] To explore the interacting effect of NT3 and LIF on the survival and initiation of primordial follicles, and investigate the possible mechanism underlying NT3 contributes to follicular development. [ Methods ] Ovaries from 4 day-old rats were cultured for 14 days in the basic medium supplemented with NT3, LIF alone or combined. Fresh （fresh control, FC） and basic medium cultured （cultured control, CC） ovaries were set as controls. The developmental stages and viability of follicles were assessed using histomorphology and TUNEL labeling. Immunohistochemistry for PCNA was performed to evaluate the mitotic activity of granulosa cells. Measurement of androstenedione and estradiol in the medium was performed and the expression of KL mRNA in the ovary was examined. [ Results ] After culture, follicles sustained healthy and less apoptotic cells were found in the presence of NT3 or LIF. Furthermore, the proportion of developmental follicles increased and the percentage of granulose cells that stained for PCNA was significantly higher. However, when combined NT3 with LIF, no obvious additional effect was found. The expression of KL mRNA in the NT3, LIF, NT3＋LIF group is 3.38±0.43, 2.02±0.20, 2.07±0.21 times higher than in the CC group. [Conclusion] NT3, LIF alone or combined had a stimulative effect on the survival and development of primordial follicles. However, there was no obvious synergistic or additional effect between LIF and NT3. One of the mechanisms of NT3 underlying the initiation of primordial follicles might activate c-kit/KL interactions.

关 键 词：NT3 LIF 卵泡发育 体外培养 大鼠 

分 类 号：R711.5[医药卫生—妇产科学]