畜牧兽医学报

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GnRH主动免疫对SD雄鼠下丘脑GnRH合成及性腺反馈系统的影响

韩兴发,曹晓涵,杜小刚,宋天增,曾宪垠*   

  1. 四川农业大学生命科学与理学院 原子能农业应用研究室, 雅安 625014
  • 收稿日期:2012-09-18 出版日期:2013-04-23 发布日期:2013-04-23
  • 通讯作者: 曾宪垠,博士生导师,主要从事动物生理与生殖免疫调控研究,Tel:0835-2886138, E-mail: xyzeng1966@163.com
  • 作者简介:韩兴发(1986-),男,四川凉山人,硕士生,主要从事动物生殖免疫调控研究,E-mail: xfhan2012@163.com;曹晓涵(1984-),女,四川成都人,博士生,主要从事动物生殖免疫调控研究,E-mail:caoxiaohan007@hotmail.com。韩兴发与曹晓涵同为第一作者
  • 基金资助:

    四川省科技厅国际合作项目(2012HH0013);四川农业大学双支计划(00770107)

Effects of Active Immunization against GnRH on the Hypothalamus GnRH Biosynthesis and Gonadal Feedback System in Sprague-Dawley Male Rats

HAN Xing-fa, CAO Xiao-han, DU Xiao-gang, SONG Tian-zeng, ZENG Xian-yin*   

  1. Isotope Research Laboratory, College of Life and Basic Sciences, Sichuan Agricultural University, Ya’an 625014, China
  • Received:2012-09-18 Online:2013-04-23 Published:2013-04-23

摘要:

探讨GnRH主动免疫对SD雄鼠下丘脑GnRH合成及性腺反馈系统的影响。36只SD雄鼠随机分为免疫组、手术去势组及对照组,免疫组于12周龄时初免, 8周后加免。放免法测定血清抗体滴度、激素浓度及下丘脑正中隆起GnRH含量,实时荧光定量PCR分析下丘脑生殖相关基因mRNA的变化。结果显示,GnRH主动免疫后12只免疫鼠中11只血清GnRH抗体滴度显著升高,血清LH、FSH 及T浓度显著降低 (P<0.05),睾丸严重萎缩(免疫去势鼠)。与对照鼠相比,免疫去势显著降低下丘脑GnRH含量(P<0.01),且显著下调下丘脑雌激素α 受体(ERα)、雄激素受体(AR)、Kiss-1、GPR54及GnRH的mRNA表达水平(P<0.05)。手术去势鼠除血清LH、FSH浓度及下丘脑ERα 的mRAN表达水平显著高于对照鼠外 (P<0.05),其余指标均与免疫去势鼠类似。结果首次表明,GnRH主动免疫抑制性腺负反馈调节作用及降低了下丘脑GnRH的合成。

Abstract:

To explore the effects of active immunization against GnRH on the hypothalamus GnRH biosynthesis and gonadal feedback regulation system in adult Sprague-Dawley (SD) rats. Thirty-six male adult SD rats at the age of 12 weeks were randomly allocated to three groups of 12 animals each. Briefly, 12 rats were immunized against GnRH with a booster vaccination 8 weeks later. 12 intact males and 12 castrated rats were not administrated and served as controls. The GnRH antibody titers and reproductive hormone levels in sera, and the content of GnRH in the median eminence were determined by radioimmunoassay (RIA). The mRNA expressions of reproduction-related genes in the hypothalamus were quantified by real-time fluorescence quantitative PCR technique. Active immunization against GnRH significantly increased serum GnRH antibody titers in eleven of twelve immunized rats, reduced serum LH, FSH and testosterone to around or undetectable levels (P<0.05), and resulted in a subcutaneously atrophy of testes in these rats (immunocastrates). Compared with intact controls, immunocastration of rats significantly reduced the content of GnRH in the median eminence and mRNA levels of androgen-receptor (AR), estrogen receptor alpha (ERα), kiss-1, GPR54 and GnRH in the hypothalamus (P<0.05). As to surgical rats, excepting the serum LH and FSH levels, and the hypothalamic ERα mRNA levels were significantly higher than those in intact rats, the remaining experimental parameters were similar to those of immunocastrates. These data firstly indicate that the feedback regulations of gonads on the hypothalamus and the synthesis capacity of GnRH in the hypothalamus are both suppressed by active immunization against GnRH.

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