摘要
目的:探讨骨形态发生蛋白4(BMP4)/Smad信号通路在小鼠原始卵泡发育中的作用及作用机制。方法:取3日龄昆明雌性小鼠卵巢体外培养,添加重组BMP4,并以不添加BMP4为对照,采用HE染色和TUNEL法,检测BMP4在原始卵泡存活和发育中的作用;采用免疫组织化学、Westernblotting与RT-PCR检测BMP4对原始卵泡中p-Smad1/5/8、sohlh2及c-kit表达的影响。结果:①BMP4组卵巢直径明显变大(1.17±0.24 mm vs 0.65±0.13 mm,P<0.05),原始卵泡数(45.3±4.2枚)明显低于对照组(55.0±5.0枚)(P<0.05),而初级卵泡的数(25.3±2.5枚)明显多于对照组(19.0±1.0枚)(P<0.05)。②TUNEL结果显示BMP4组凋亡卵母细胞比例明显低于对照组(P<0.05)。③免疫组织化学、Western blotting与RT-PCR结果显示,与对照组相比,BMP4组原始卵泡卵母细胞中p-Smad 1/5/8、Sohlh2及C-kit的表达均显著升高(P<0.01)。结论:BMP4/Smad信号通路可促进小鼠原始卵泡向初级卵泡转变,并抑制卵母细胞的凋亡;其可能通过上调Sohlh2及C-kit基因的表达而发挥作用。
Objective： To explore the effects and mechanism of bone morphologenetic protein 4 （BMP4）/ Smad signaling pathway in mouse primordial follicle development. Methods： Ovaries from 3-day-old Kunming mouse were cultured in vitro, and randomly divided into two groups, BMP4 （final concentration 100 ng/ml） group and the control （without BMP4）. HE staining and TUNEL were used to examine the effects of BMP4 on the primordial follicle survival and development. The mRNA and protein expression of p-Smadl/5/8, sohlh2 and c-kit were investigated by immunohistochemistry, Western blotting and RT-PCR. Results： 1） Ovaries in BMP4 group were much bigger than those in the control （1.17 ± 0.24 mm vs 0.65 ± 0.13 mm, P〈0.05）; there were more primary follicles and less primordial follicles in BMP4 group （19.0 ± 1.0, 55.0 ±5.0） than those in the control （25.3 ± 2.5, 45.3 ± 4.2） （P〈0.05）; 2） TUNEL results demonstrated that the ratio of apoptotic oocyte in BMP4 group was significantly lower than that in the control （P〈0.05）. 3） Immunohistochemistry, RT-PCR and Western blotting results revealed that BMP4 increased the expression of p-Smadl/5/8, Sohlh2 and C-kit in the oocytes of primorial follicles. Conclusion： BMP4/Smad signaling pathway could promote the primordial-to-primary folicle transition and inhibit oocyte apoptosis in mouse, by up-regulating the expression of Sohlh2 and c-kit.
出处
《生殖与避孕》
CAS
CSCD
2012年第10期649-655,680,共8页
Reproduction and Contraception
基金
国家自然科学基金课题(No.31071269)
山东大学杰出青年培育基金课题(2010JQ003)
山东省人口和计划生育科研基金课题(200902)
