抗缪勒氏管激素抑制山麻鸭卵泡发育分子机制的研究

doi:10.16431/j.cnki.1671-7236.2023.08.021

摘要/Abstract

摘要： 【目的】探究抗缪勒氏管激素(anti-Müllerian hormone,AMH)抑制山麻鸭卵泡发育的分子调控机制。【方法】选取40只29周龄产蛋山麻鸭,随机分为4组:AMH1、AMH2、AMH3和对照组,AMH1、AMH2和AMH3组给予2、20和200 μg/d AMH多肽;对照组给予同等体积的生理盐水,整个试验持续21 d;每天记录产蛋量,在试验末采集卵巢和卵泡进行计数,称重计算卵巢指数,测定血清促黄体生成素(luteinizing hormone,LH)、孕酮(progesterone,P₄)和雌二醇(estradiol,E₂)水平,检测等级卵泡(F6)、小黄卵泡(SYF)和大白卵泡(LWF)中卵泡刺激素受体(FSHR)、促黄体生成素受体(LHR)、类固醇生成急性调节蛋白(StAR)、3β-羟类固醇脱氢酶(3β-HSD)、雌激素合成酶(CYP19A1)、生长分化因子9(GDF9)、骨形态发生蛋白15(BMP15)基因表达量,以及F6和SYF中StAR、GDF9蛋白表达水平。【结果】与对照组相比,AMH2和AMH3组的产蛋率显著下降(P＜0.05),其中AMH3组对产蛋率的抑制作用最为明显;AMH1和AMH2组SYF和LWF数量显著上升(P＜0.05),而卵巢指数和LYF数量没有明显变化,血清LH、P₄和E₂水平呈剂量依赖性下降,AMH3组对生殖激素抑制作用较明显,其中AMH3组LH水平显著下降(P＜0.05)。与对照组相比,在F6中,AMH2组GDF9基因和AMH1组BMP15基因表达水平显著上升(P＜0.05);在SYF中,除AMH2组GDF9基因外,各试验组FSHR基因表达水平显著下降,GDF9和BMP15基因表达水平显著上升(P＜0.05),AMH3组GDF9蛋白水平显著上升(P＜0.05);在LWF中AMH2和AMH3组的GDF9和BMP15基因表达水平显著上升(P＜0.05)。给予外源性AMH对F6和LWF中CYP19A1、3β-HSD和StAR基因表达水平均无显著影响(P＞0.05),在SYF中,AMH1、AMH2和AMH3组中CYP19A1、3β-HSD和StAR基因表达水平均显著低于对照组(除AMH2组的CYP19A1基因外,P＜0.05);与对照组相比,给予外源性AMH对F6和SYF中StAR的蛋白水平均无显著影响(P＞0.05)。【结论】200 μg/d外源性AMH可显著抑制卵泡发育,主要通过抑制LH、P₄和E₂的分泌,上调卵泡中GDF9和BMP15表达水平,下调FSHR和LHR以及影响类固醇激素合成通路关键因子的表达来影响卵泡发育。

关键词: 抗缪勒氏管激素(AMH); 促性腺激素受体; 类固醇; 卵泡发育; 山麻鸭

Abstract: 【Objective】 Investigating the inhibition of anti-Müllerian hormone (AMH) on follicular development and the molecular mechanism in Shan-ma ducks.【Method】 40 healthy 29-week-old Shan-ma ducks were randomly divided into 4 groups.AMH1,AMH2 and AMH3 groups were given 2,20 and 200 μg/d AMH for 21 d,respectively.And the control group was given saline.During the experiment,egg production was recorded daily.Samples were collected at the end of this experiment,the number of follicles were counted,and calculated the gonadosomatic index (GSI).Serum luteinizing hormone (LH),progesterone (P₄) and estradiol (E₂) levels,the gene expression levels of Follicle stimulating hormone receptor (FSHR),luteinizing hormone receptor (LHR),steroidogenic acute regulatory protein (StAR),3β-hydroxysteroid dehydrogenase (3β-HSD),estrogen synthase (CYP19A1),growth differentiation factor 9 (GDF9),bone morphogenetic protein 15 (BMP15) in graded follicles (F6),small yellow follicles (SYF) and large white follicles (LWF),and the protein levels of StAR and GDF9 in F6 and SYF were detected.【Result】 Compared with control group, the laying rate were significantly decreased in AMH2 and AMH3 groups (P＜0.05),and AMH3 group showed the most significant inhibition of egg production rate. The numbers of SYF and LWF were significantly increased in AMH1 and AMH2 groups (P＜0.05), while GSI and the numbers of LYF had no significant trend. Serum LH, P4 and E2 level were in dose-dependent reduction, and AMH3 group showed a more pronounced inhibition of these hormones. LH level was significantly decreased in AMH3 group (P＜0.05). Compared with control group, the expression of GDF9 gene in AMH2 group and BMP15 gene in AMH1 group were significantly increased in F6 (P＜0.05). In SYF, except for GDF9 gene in the AMH2 group, the expression of FSHR gene was significantly decreased, while the expression of GDF9 and BMP15 genes in each experiment group were significantly increased (P＜0.05). The GDF9 protein level in AMH3 group was significantly increased (P＜0.05). The expression of GDF9 and BMP15 genes in AMH2 and AMH3 groups were significantly increased in LWF (P＜0.05). The administration of exogenous AMH did not significantly affect the expression of CYP19A1,3β-HSD and StAR genes in F6 and LWF (P＞0.05). In SYF, CYP19A1,3β-HSD and StAR genes were observed in AMH1, AMH2, and AMH3 groups were significantly lower than those of control group (except for CYP19A1 gene in AMH2 group, P＜0.05). Compared with control group, the administration of exogenous AMH had no significant effect on the protein levels of StAR in F6 and SYF (P＞0.05).【Conclusion】 200 μg/d exogenous AMH could significantly inhibit follicular development,mainly by inhibiting the secretion of LH,P₄,and E₂,upregulating the expression of GDF9 and BMP15 in follicles,downregulating the expression of FSHR and LHR,as well as key factors affecting steroid hormone synthesis pathways.

Key words: anti-Mullerian hormone(AMH); gonadotropin receptor; steroid hormone; follicular development; Shan-ma ducks

中图分类号:

S859

周晓莉, 张焯深, 沈栩, 潘建秋, 黄运茂, 江丹莉. 抗缪勒氏管激素抑制山麻鸭卵泡发育分子机制的研究[J]. 中国畜牧兽医, 2023, 50(8): 3239-3247.

ZHOU Xiaoli, ZHANG Zhuoshen, SHEN Xu, PAN Jianqiu, HUANG Yunmao, JIANG Danli. Study on the Molecular Mechanism of Inhibition of Follicular Development in Shan-ma Ducks by Anti-Müllerian Hormone[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3239-3247.

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