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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
Effects of CART on Estradiol Production in vitro in Follicular Granulose Cells of Sheep Ovarian
【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 stim-ulating 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7d,分别在培养48、96、144和168h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50ng·mL-1),细胞培养168h后,雌激素浓度显著提高。当FSH浓度为5ng·mL-1时,培养液雌激素浓度最高为5 694.5±40.5pg·mL-1;当FSH浓度为5ng·mL-1,随着CART不同浓度的加入(0.01、0.10、1.00μmol.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、50ng·mL-1水平下加入CART浓度为0.10μmol.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.
【Key words】 sheep; CART; FSH; granulosa cell; culture in vitro; estradiol;
- 【文献出处】 畜牧兽医学报 ,Chinese Journal of Animal and Veterinary Sciences , 编辑部邮箱 ,2013年06期
- 【分类号】S826
- 【被引频次】13
- 【下载频次】146