可卡因-苯丙胺调控转录肽(CART)对猪卵巢卵泡颗粒细胞雌激素产生的影响

摘要/Abstract

摘要：

旨在研究可卡因苯丙胺调控转录肽（Cocaine amphetamine regulated transcript, CART）对促卵泡素(Follicular stimulating hormone, FSH)诱导的猪卵巢卵泡颗粒细胞雌激素产生的影响。用Long-term培养方法在不同CART、FSH浓度下对猪卵巢卵泡颗粒细胞培养7 d，在显微镜下观察各孔细胞生长情况并采集图像；采用双抗体夹心ABC-ELISA法测定各孔雌激素浓度。细胞培养168 h后：（1）在培养液不添加CART时，添加FSH孔中的雌激素浓度与不添加孔相比，显著提高；浓度为25 ng·mL^-1时，雌激素浓度最高，为(17 295.57±184.03) pg·mL^-1；（2）CART对体外培养的无FSH和FSH为5 ng·mL^-1的猪卵巢卵泡颗粒细胞雌激素的产生没有影响；（3）当培养体系FSH浓度分别为25 和50 ng·mL^-1时，随着CART浓度的升高（0.01、0.1、1 μmol·L^-1），培养液雌激素浓度有下降的趋势，但各组间差异不显著（P>0.05）。但是，当FSH 为25 ng·mL^-1，CART为 0.1和1 μmol·L^-1时，与对照组相比，颗粒细胞雌激素的产生明显受到抑制（P<0.05）。但是，统计分析FSH和CART互作效应并不显著（P>0.05），这一点需进一步试验证明；（4）当培养液添加50 ng·mL^-1FSH时，随着CART浓度的升高（0.01、0.1、1 μmol·L^-1），与对照组相比，雌激素浓度各组差异不显著（P>0.05）。猪卵巢卵泡颗粒细胞雌激素的产生需在FSH诱导下进行；当培养液FSH为25和50 ng·mL^-1时，CART对FSH诱导的猪卵巢卵泡颗粒细胞雌激素的产生有抑制作用，但抑制效果并不显著。由此推断，并不像单胎的牛、羊，CART在猪卵泡发育过程中，可能并非是个主要的局部负调控因子。

Abstract:

The study focused on the effect of CART on FSH-induced estradiol production of porcine ovarian follicular granulose cells.Granulose cells were cultured for 7 days under different CART and FSH concentrations by Long-term culture. The growth situation in each well was observed and pictures were taken. The content of estradiol in each well was determined by sandwich ABC-ELISA. After 168 h of incubation, (1) In CART-free medium, the estrodiol levels in wells supplemented with FSH significantly increased than that of FSH-free well. Adding of 25 ng·mL^-1 of FSH resulted in the highest estrodiol level of (17 295.57±184.03) pg·mL^-1; (2) CART showed no effect on estradiol production by porcine ovarian follicular granulose cells cultured in FSHfree and 5 ng·mL^-1 FSH medium; (3) In the medium containing 5 or 25 ng·mL^-1 FSH,with the increase of CART concentration（0.01, 0.1, 1 μmol·L^-1）, the levels of estradiol concentration showed a tendency to decrease but there was no significant differences among the three groups(P>0.05). However, when FSH at 25 ng·mL^-1, CART at 0.1 or 1 μmol·L^-1, compared with the control group (FSH 25 ng·mL^-1, CART 0 μmol·L^-1), estradiol secreted by follicular granulose cells was suppressed markedly(P<0.05). The result of statistical analysis demonstrated no significant interactive effect between FSH and CART (P>0.05), which needs further investigation. (4) When the medium contained 50 ng·mL^-1 of FSH, the increase of CART concentration showed no significant influence on estradiol concentration in different groups (P>0.05). The production of follicular estradiol in pig was induced by FSH ; when the concentration of FSH were 25 and 50 ng·mL^-1, CART inhibited the estradiol production by porcine follicular granulosa cells, but no significant inhibitory effects.The result indicate that CART maybe not a main local negative regulatory factor in pig follicular development.

中图分类号:

S828
S814.1

李鹏飞，岳文斌，李富禄，黄洋，孙晋艳，朱芷薇，于秀菊，贺俊平，范瑞文，任有蛇，吕丽华. 可卡因-苯丙胺调控转录肽(CART)对猪卵巢卵泡颗粒细胞雌激素产生的影响[J]. 畜牧兽医学报, doi: .

LI Peng-fei,YUE Wen-bin, LI Fu-lu, HUANG Yang, SUN Jin-yan, ZHU Zhi-wei, YU Xiu-ju, HE Jun-ping, FAN Rui-wen, REN You-she, LV Li-hua. Effects of CART on Estradiol Production of Pig Ovarian Follicular Granulosa Cells in vitro Culture[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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