CART对FSH诱导下水牛颗粒细胞雌二醇分泌及相关基因表达的影响

doi:10.16431/j.cnki.1671-7236.2020.09.021

Abstract

Abstract: In this study,primary buffalo ovarian granulosa cells were cultured in a serum-free culture medium in vitro to investigate the effect of exogenous cocaine amphetamine regulated transcript (CART) on the production of estradiol (E₂) in granulosa cells induced by follicle stimulating hormone (FSH) and its related gene expression.The granulosa cells was cultured for 7 days by using a long-term method,in each group that was adding different concentration levels of FSH (0,0.25,0.5,1,2 and 4 ng/mL).The concentration of E₂ was assayed by ABC-ELISA kit to screen out the optimal dosage of FSH.Primary granulosa cells were randomly divided into 4 group (2 groups of control (without FSH) and 2 groups of optimal FSH levels) for 6 days culure,one of the control group and FSH group were respectively combined with 25 μg/mL CART for 24 h,to detect the change of E₂ concentration and quantitative detection the gene expression of CART receptors and aromatase in granulosa cell.The results showed as the follows:①Without CART,the E₂ concentration in granulose cells increased first and than decreased with the increased of FSH.When adding 1 ng/mL FSH,the concentration of E₂ reached the hightest (116.16 pmol/mL) and significantly higher than the other groups (P<0.05);②Adding CART significantly reduced the concentration of E₂ in granulosa cells induced by FSH (P<0.05) and the two control groups 0 FSH and 0 FSH+CARD had no significantly difference (P>0.05).③With 1 ng/mL FSH,exogenous CART significantly reduced the expression of CYP19A1 mRNA in granulosa cells after 24 h treated (P<0.05),while the relative expression of CARTPT mRNA in granulosa cells significantly increased (P<0.05).Under the condition of 0 ng/mL FSH,the expression of CYP19A1 and CARTPT mRNA had no significant difference (P>0.05).Therefore,this study was speculated that CART was a negative regulatory signal in the ovarian development of buffalo,which inhibited the healthy development of follicles,a certain amount of FSH was required to induced the negative pressure of CART on granulosa cell proliferation and differentiation.

Key words: CART; FSH; E₂; buffalo; granulose cells; CYP19A1

CLC Number:

S814.4

LI Lingyu, ZHENG Haiying, YU Nongqi, SHANG Jianghua, Huang Jiaxiang, Yang Chunyan. Effects of CART on Estradiol and Related Gene Expression in Buffalo Follicle Granulosa Cells Induced by FSH[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2876-2883.

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Effects of CART on Estradiol and Related Gene Expression in Buffalo Follicle Granulosa Cells Induced by FSH

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