畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (6): 853-857.doi: 10.11843/j.issn.0366-6964.2013.06.004

• 遗传繁育 • 上一篇    下一篇

可卡因-苯丙胺调控转录肽对绵羊卵巢卵泡颗粒细胞雌激素产生的影响

李鹏飞1,岳文斌2,黄洋2,孙晋艳2,李晓明2,庞钰莹2,于学静2,贺俊平2,孟金柱2,任有蛇2,吕丽华2*   

  1.  (1.山西农业大学生命科学学院,太谷 030801; 2.山西农业大学动物科技学院,太谷 030801)
  • 收稿日期:2012-11-19 出版日期:2013-06-23 发布日期:2013-06-23
  • 通讯作者: 吕丽华, E-mail:sxaullh@yahoo.com.cn
  • 作者简介:李鹏飞(1978-),男,山西偏关人,讲师,博士,主要从事动物生殖生理方面的研究,E-mail:adamlpf@126.com
  • 基金资助:

    国家自然科学基金(31172211);农业部948项目(2010-Z43);山西省留学基金(2008080);山西省国际科技合作项目(2010081002);山西农业大学新增横向协作与委托项目(2010HX54);国家重点实验室开放课题(SNKF-2012-02);山西省科技攻关项目(20130311027-2)

Effects of CART on Estradiol Production in vitro in Follicular Granulose Cells of Sheep Ovarian

LI Peng-fei1, YUE Wen-bin2, HUANG Yang2, SUN Jin-yan2, LI Xiao-ming2, PANG Yu-ying2, YU Xue-jing2, HE Jun-ping2, MENG Jin-zhu2, REN You-she2, LYU Li-hua2*   

  1. (1. College of Life Science, Shanxi Agricultural University, Taigu 030801, China; 2. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China
  • Received:2012-11-19 Online:2013-06-23 Published:2013-06-23

摘要:

研究可卡因-苯丙胺调控转录肽(Cocaine amphetamine regulated transcript,CART)对促卵泡素(Follicular stimulating hormone,FSH)诱导的绵羊卵巢卵泡颗粒细胞雌激素(E2)产生的影响。收集卵巢卵泡颗粒细胞,Long-term培养方法在不同CART、FSH浓度下培养7 d,分别在培养48、96、144和168 h后,在显微镜下观察各孔细胞生长情况;采用双抗体夹心ABC-ELISA法测定其雌激素浓度。培养液分别加入不同浓度FSH(5、25和50 ng·mL-1),细胞培养168 h后,雌激素浓度显著提高。当FSH浓度为5 ng·mL-1时,培养液雌激素浓度最高为5 694.5±40.5 pg·mL-1;当FSH浓度为5 ng·mL-1,随着CART不同浓度的加入(0.01、0.10、1.00 μmol·L-1),颗粒细胞雌激素的产生受到不同程度的抑制;在FSH浓度为25、50 ng·mL-1水平下加入CART浓度为0.10 μmol·L-1时,对FSH诱导的绵羊卵巢卵泡颗粒细胞雌激素产生的抑制作用明显(P<0.01)。绵羊卵巢卵泡颗粒细胞雌激素的产生需在FSH诱导下进行;CART对FSH诱导的绵羊卵巢卵泡颗粒细胞雌激素的产生有抑制作用。

Abstract:

 The objective of this study is to elucidate effects of CART on estradiol (E2) production in vitro in follicular granulosa cells of sheep ovaries. The ovarian granulosa cells were cultured for 7 days by using the Long-term method. The cell growth status in each well was observed under a microscope after cultured of 48, 96, 144 and 168 h, respectively under different concentrations of CART and FSH. The concentration of estradiol was assayed by ABC-ELISA. After 168 h of culture, culture medium with different concentrations of FSH (5, 25 and 50 ng·mL-1) was added, the concentration of estradiol was significantly increased. When adding 5 ng·mL-1 FSH, the concentration of estradiol was the highest (5 694.5±40.5 pg·mL-1). When the concentrations of FSH was 5 ng·mL-1, with the different concentrations of CART (0.01, 0.10 and 1.00 μmol·L-1) was added, the production of estradiol in granulosa cells were inhibited in different degrees. When the concentrations of FSH was 25 and 50 ng·mL-1 respectively, and CART was 0.10 μmol·L-1 in the culture media, the inhibition was significant for the production of estradiol in FSH-induced sheep ovarian granulosa cells. The generation of estradiol in sheep ovarian granulosa cells needs FSH induction. CART inhibits the estradiol production of FSH-induced sheep follicular granulosa cells in vitro.

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