原癌基因在介导细胞因子影响卵母细胞成熟中的作用  

THE EFFECTS OF PROTOONCOGENE ON OOCYTE MATURATION MEDIATED BY CYTOKINES

作　　者：郑莉萍[1] 王晶磊[1] 郑月慧[1] 吴磊[1] 肖秋香[2] 李芳[1] 

机构地区：[1]南昌大学医学院生殖研究室,江西南昌330006 [2]江西省赣南医学院,江西赣州341000

出　　处：《中国应用生理学杂志》2009年第1期74-79,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(30260035);江西省自然科学基金资助项目(2007GCY1001)

摘　　要：目的:探讨原癌基因c-erbB_2、c-myb在介导表皮生长因子、肿瘤坏死因子α、内皮素-1及一氧化氮影响卵母细胞成熟中的作用。方法:建立小鼠卵母细胞体外培养模型,观察EGF、TNFα、ET-1及NO供体小剂量硝普钠(SNP)对卵细胞成熟的影响,用RT-PCR和Western blotting方法检测在上述细胞因子作用下卵母细胞c-erbB2和c-myb表达及丝裂原蛋白激酶(MAPK)和成熟促进因子(MPF)含量的变化。结果:EGF(10μg/L)能促进卵母细胞的成熟,使c-erbB_2 mRNA、c-myb mRNA表达上升,伴随MAPK磷酸化水平和cyclinB1含量升高;TNFα(10μg/L)和ET-1(10^(-7)mol/ L)均使卵母细胞中c-erb_2 mRNA和c-myb mRNA表达下降,伴随MAPK磷酸化水平和cyclinB1含量下降,抑制卵母细胞的成熟。小剂量SNP(10^(-5)mol/L)对卵母细胞成熟无明显影响,但能反转二丁酰环磷酸腺苷(dbcAMP)对卵母细胞成熟的抑制作用,使c-erbB_2mRNA和c-myb mRNA表达升高,MAPK磷酸化水平及cyclinB1含量升高,但仍低于对照组。结论:细胞因子对卵母细胞成熟的影响与原癌基因c-erbB_2和c-myb有关,c-erbB_2和c-myb可能是介导细胞因子和MAPK、MPF间调控卵母细胞成熟的上游激活物。Aim： The mechanisms of cytokines in regulating oocyte maturation is still little known. The present study attempt to investigate whether the protooncogene of c-erbB2, c-myb are invoked in introducing of cytokines to regulate oocyte maturation. Methods： This research used mouse GV stage oocyte culture model in vitro and RT-PCR, Western blotting method to explore the effect of EGF, TNFα, ET-1 and NO on oocyte maturation; to analyze the c-erbB2 mRNA and c-myb mRNA expression and the phosphorylation of MAPK and cyclinB1 expression in oocytes affected by above cytokines. Results： EGF（10 μg/L） stimulated meiosis of oocytes significantly, the level of c-erbB2 mRNA, c-myb mRNA were increased, and promoted the phosphorylation of MAPK and cyclinB1 expression; TNFα（ 1 μg/L） and ET-1 （（ 10^-1 mol/L） had the contrary results to EGF. Low dose of SNP（（10^-5mol/L） had no effect on oocyte maturation, but could significantly reverse the suppression of dbcAMP on oocyte maturation. Conclusion： c-erbB2 and c-myb were involved in introducing of cytokines to regulate oocyte maturation, might be the middle link in connection of the cytokines with MAPK and MPF in regulation oocyte maturation.

关 键 词：卵母细胞成熟 c—erbB2和c—myb 细胞因子 MAPK和MPF 

分 类 号：R335.5[医药卫生—人体生理学]