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目的在小鼠卵巢玻璃化冻融过程中给予重组人卵泡雌激素(r FSH)干预,通过观察其对卵泡刺激素受体(follicle-stimulating hormone receptor,FSH-R)及与血管生成相关的Integrinαvβ3、Angpt-2、Kindlin-2蛋白表达的影响,提供FSH干预对卵巢血管生成相关因子影响的依据。方法将4周龄C57BL/6J小鼠卵巢随机分为新鲜对照组(FCG)、玻璃化冻存对照组(VCG)、0.3IU·m L-1FSH干预玻璃化冻存组(FSH-VG)。对3组卵巢内各级卵泡进行计数;通过免疫组化方法观察并分析各组FSH-R、Angpt-2、Kindlin-2在卵巢不同细胞中的定位;观察粘附分子Integrinαvβ3在各组卵巢细胞中表达,并进行亚基表达量的分析。结果FSH-VG组的原始卵泡、初级卵泡、正常卵泡及总卵泡数均高于VCG组,但低于FCG组(P均<0.05),闭锁卵泡数VCG组高于FCG及FSH-VG组(P均<0.05);FSH-R与Angpt-2阳性表达呈现FSH-VG与VCG组均低于FCG组(P<0.05),但FSH-VG组高于VCG组(P<0.05);Kindlin-2阳性表达在FSH-VG组与FCG组之间差异无统计学意义(P>0.05),但均高于VCG组(P<0.01或<0.05);Integrinβ3蛋白表达量FCG与FSH-VG组间差异无统计学意义(P>0.05),但均高于VCG组(P<0.05),Integrinαv的表达3组间差异无统计学意义(P>0.05)。结论玻璃化冻融过程中添加FSH可以上调FSH-R及粗血管生成的相关因子Integrinβ3、Angpt-2和Kindlin-2蛋白的表达,提高正常卵泡数。
Objective To observe the effect of r FSH on the follicle-stimulating hormone receptor (FSH-R) and angiogenesis-related Integrinαvβ3 , Angpt-2, Kindlin-2 protein expression, provide basis for FSH intervention on ovarian angiogenesis-related factors. Methods Four-week-old C57BL / 6J mice were randomly divided into three groups: fresh control group (FCG), vitrified control group (VCG) and 0.3IU · m L-1 FSH intervention group (FSH-VG). The ovarian follicles at all levels were counted in three groups. The expression of FSH-R, Angpt-2 and Kindlin-2 in different ovarian cells was observed by immunohistochemistry and the expression of integrinαvβ3 in ovarian cells In expression, and subunit expression analysis. Results The number of primary follicles, primary follicles, normal follicles and total follicles in FSH-VG group were higher than those in VCG group, but lower than those in FCG group (all P <0.05). The number of atresia follicles in VCG group was higher than that in FCG group and FSH-VG group P <0.05). The positive expression of FSH-R and Angpt-2 in FSH-VG and VCG groups was lower than that in FCG group (P <0.05), but higher in FSH-VG group than in VCG group (P <0.01 or <0.05). The expression of integrinβ3 in FCG group and FSH-VG group was significantly higher than that in VCG group (P <0.01 or <0.05). There was no significant difference between FSH-VG group and FCG group There was no significant difference between the two groups (P> 0.05), but both of them were higher than VCG group (P <0.05). There was no significant difference in the expression of Integrinαv between the three groups (P> 0.05). Conclusion FSH-R and the expression of integrinβ3, Angpt-2 and Kindlin-2, which are related factors of crude angiogenesis, can be increased by adding FSH during vitrification and freeze-thaw cycles to increase the number of normal follicles.