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

摘要

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 168h 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 1.00μmol · 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 0.10μmol · 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.%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5694.5±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.