摘要
目的:探讨激活素-抑制素-卵泡抑素(ACT-INH-FS)系统在小鼠生发泡(GV)期卵母细胞体外成熟及早期胚胎发育中的的影响机制。方法:取小鼠GV期卵母细胞,随机分成8组,A组:100ng/ml激活素A(ACTA);B组:100ng/mlACTA+100ng/ml卵泡抑素(FS);C组:100ng/ml抑制素A(INHA);D组:100ng/mlINHA+100ng/mlFS;E组:100ng/mlACTA+INHA;F组:100ng/mlACTA+100ng/mlINHA+100ng/mlFS;G组:100ng/mlFS;H组(对照组):人输卵管液(HTF)+10%血清蛋白代用品(SPS)。体外培养16-18h后,MⅡ期卵母细胞进行体外受精,观察ACTA-INHA-FS对GV期卵母细胞生发泡破裂(GVBD)、第一极体(PB1)排出及受精率、卵裂率、囊胚形成率的影响。使用MitoTrackerRed荧光探针对A组及E组MⅡ期卵母细胞内活性线粒体数目及分布进行检测。结果:与H组比,A、E、G组GVBD发生率、A、C、E组的MⅡ期卵母细胞发生率和4-细胞、8-细胞及囊胚形成率均显著增加,C组受精率下降,P均<0.05。A、E组卵母细胞内线粒体形态半周型及弥散型比例明显高于H组,整体分布存在显著性差异(P<0.01)。结论:①ACTA、INHA对GV期卵母细胞体外成熟及胚胎发育潜能具有显著的促进作用,并可以被FS所拮抗。②在ACTA/INHA缺乏或不足的条件下,FS可出现ACTA样作用,促进卵母细胞成熟。③ACTA-INHA对体外成熟卵母细胞核质成熟同步化具促进作用。
Objective： To investigate the effect of ACT-INH-FS on in vitro maturation and development potential of GV （germinal vesicle） stage mouse oocyte. Methods： 1） Mice oocytes at GV stage were matured for 16- 18 h in control media （HTF＋ 10%SPS, group H） and 7 treatment media （group A： 100 ng/rnl ACTA, group B： 100 ng/ml ACTA＋ 100 ng/ml FS, group C： 100 ng/ml ACTA, 100 ng/ml INHA＋100 ng/ml FS, group D： 100 ng/ml ACTA＋100 ng/ml INHA, group E： 100 ng/ml ACTA＋100 ng/ml INHA＋100 ng/ml FS, group F： 100 ng/ml FS, respectively）. Oocytes at MII stage were fertilized and cultured using standard procedures. The germinal vesicle break down（GVBD）, polar body l（PB1）exclusion rate, fertilization rate and the frequency of blastocyst formation were compared. 2） Oocytes at MII stage matured in either group A or group E were stained with Mito-Tracker Red. Active mitochondria number and relocation were analyzed. Results： Compared with group H, the GVBD rates of group A, E, G were improved obvious.ly; Oocytes at MII stage and the 4-cell, 8-cell and blastocyst formation rate of group A, C, E were significantly higher; the fertilization rate of group C was lower; P all 〈0.05. In group A and E, much more oocytes presented a semiperipheral pattern or a diffused distribution of active mitochondria （P〈0.05）. Conclusions： 1） INHA and ACTA exert development-enhancing effects during in vitro maturation and early embryo development. 2） When ACTA is deficient or absent, FS may play an ACT-like role. 3） ACTA and INHA could improve the synchronism in the progression of nuclear and cytoplasmic maturation.
出处
《生殖与避孕》
CAS
CSCD
北大核心
2008年第3期134-139,共6页
Reproduction and Contraception
