雄激素受体对绵羊卵巢颗粒细胞发育的调控作用

doi:10.11843/j.issn.0366-6964.2026.01.021

畜牧兽医学报 ›› 2026, Vol. 57 ›› Issue (1): 246-257.doi: 10.11843/j.issn.0366-6964.2026.01.021

雄激素受体对绵羊卵巢颗粒细胞发育的调控作用

李容青¹(), 特日格勒³, 付绍印³, 何小龙³, 达赖³, 王力伟³, 高元帅¹, 张文佳¹, 解志强¹, 杨会国⁴, 王标³(), 刘永斌²()

^1.内蒙古大学，呼和浩特 010000
^2.内蒙古农业大学，呼和浩特 010000
^3.内蒙古自治区农牧业科学院，呼和浩特 010000
^4.新疆维吾尔自治区畜牧科学院，乌鲁木齐 830011

收稿日期:2025-05-23 出版日期:2026-01-23 发布日期:2026-01-26
通讯作者: 王标,刘永斌 E-mail:rong-qing.li@mail.imu.edu.cn;wangbnd@163.com;ybliu117@126.com
作者简介:李容青，博士，主要从事绵羊遗传资源与干细胞生物学研究，E-mail：rong-qing.li@mail.imu.edu.cn
基金资助:
国家重点研发计划(2021YFD1300904);内蒙古自治区揭榜挂帅项目(2022JBGS0012);内蒙古自治区揭榜挂帅项目(2022JBGS0024);内蒙古自治区科技计划项目(2019GG349);内蒙古自治区科技计划项目(2021GG0443);现代农业产业技术体系专项资金(CARS-38)

The Regulatory Role of Androgen Receptor in the Development of Ovarian Granulosa Cells in Sheep

LI Rongqing¹(), TE Rigele³, FU Shaoyin³, HE Xiaolong³, DA Lai³, WANG Liwei³, GAO Yuanshuai¹, ZHANG Wenjia¹, XIE Zhiqiang¹, YANG Huiguo⁴, WANG Biao³(), LIU Yongbin²()

^1.Inner Mongolia University，Hohhot 010000，China
^2.Inner Mongolia Agricultural University，Hohhot 010000，China
^3.Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences，Hohhot 010000，China
^4.Xinjiang Uygur Autonomous Region Academy of Animal Science，Urumqi 830011，China

Received:2025-05-23 Online:2026-01-23 Published:2026-01-26
Contact: WANG Biao, LIU Yongbin E-mail:rong-qing.li@mail.imu.edu.cn;wangbnd@163.com;ybliu117@126.com

摘要/Abstract

摘要：

旨在探究敲低雄激素受体（androgen receptor，AR）后对绵羊卵巢颗粒细胞（granulosa cells，GC）发育的影响机制。本研究选取健康成年绵羊为试验对象，手术法采集卵巢，分离并鉴定绵羊卵巢颗粒细胞。敲低绵羊卵巢颗粒细胞的AR，通过CCK-8法、ELISA法检测绵羊卵巢颗粒细胞的增殖活性和雌二醇（estradiol，E2）的分泌水平。敲低AR后通过转录组测序（transcriptome Sequencing，RNA-seq）筛选影响绵羊卵巢颗粒细胞发育的关键差异表达基因（DEGs），对其进行功能富集分析，并利用RT-qPCR和Western blot验证与绵羊卵巢颗粒细胞发育相关的信号通路及其差异基因。结果表明，本研究成功分离并鉴定了绵羊卵巢颗粒细胞；敲低AR后会显著降低绵羊卵巢颗粒细胞的增殖活性。当AR被敲低后，通过转录组测序筛选到1 187个差异基因，465个基因上调，722个基因下调。根据这些差异基因筛选出对绵羊卵巢颗粒细胞发育具有重要调控作用的关键信号通路：PI3K-Akt信号通路和雌激素信号通路，并对关键基因PI3K、AKT、p-AKT、ESR2、CYP19A1、BAX、BAX/BCL2、Caspase3进行验证；敲低AR后PI3K-Akt信号通路中关键基因PI3K、AKT、p-AKT显著下调（P<0.05），雌激素合成关键基因ESR2、CYP19A1显著下调（P<0.05），而细胞增殖相关基因BAX、BAX/BCL2、Caspase3显著上调（P<0.05）。综上所述，本研究发现敲低绵羊卵巢颗粒细胞AR后，可能通过PI3K-Akt信号通路和雌激素信号通路来影响绵羊卵巢颗粒细胞的增殖和激素合成，从而参与细胞的发育过程。

关键词: AR, 颗粒细胞, 增殖, 激素, 发育

Abstract:

This study aimed to investigate the mechanism by which knockdown of androgen receptor （AR） in sheep affects the development of ovarian granulosa cells （GC）. The healthy， adult sheep was selected as experimental subjects， employing surgical procedures to obtain ovarian tissue， from which granulosa cells were subsequently isolated and characterized.After silencing AR expression in granulosa cells （GCs）， cell proliferation and estradiol （E2） production were assessed in sheep ovarian GCs through CCK-8 and ELISA techniques.Furthermore， after AR knockdown， key differentially expressed genes （DEGs） affecting ovarian granulosa cell development were identified through transcriptome sequencing （RNA-seq）. These genes were subjected to functional enrichment analysis， and signaling pathways involved in granulosa cell development， along with their associated DEGs， were validated using RT-qPCR and Western blot assays.The results demonstrated that the study successfully isolated and identified granulosa cells， and that knockdown of AR significantly reduces the proliferation activity of these cells.More importantly， after AR was knocked down， 1 187 differentially expressed genes was identified through transcriptome sequencing， with 465 genes upregulated and 722 genes downregulated.Based on these differentially expressed genes， key signaling pathways involved in granulosa cell development were identified， including the PI3K-Akt pathway and the estrogen signaling pathway. The key genes PI3K， AKT， p-AKT， ESR2， CYP19A1， BAX， BAX/BCL2， and Caspase3 were further validated.Following AR silencing， the expression levels of critical genes within the PI3K-Akt signaling cascade—specifically PI3K， AKT， and p-AKT were significantly reduced （P<0.05）. Similarly， key genes associated with estrogen biosynthesis， ESR2 and CYP19A1， exhibited significant downregulation （P<0.05）. Conversely， genes linked to cellular proliferation， including BAX， the BAX/BCL2 ratio， and Caspase3， showed a significant increase in expression （P<0.05）. In summary， these findings suggest that knocking down AR in sheep ovarian granulosa cells may influence cell proliferation and hormone synthesis through the PI3K-Akt signaling pathway and the estrogen signaling pathway， thereby participating in the process of cellular development.

Key words: AR, granular cells, proliferation, hormone, development

中图分类号:

S826.3

李容青, 特日格勒, 付绍印, 何小龙, 达赖, 王力伟, 高元帅, 张文佳, 解志强, 杨会国, 王标, 刘永斌. 雄激素受体对绵羊卵巢颗粒细胞发育的调控作用[J]. 畜牧兽医学报, 2026, 57(1): 246-257.

LI Rongqing, TE Rigele, FU Shaoyin, HE Xiaolong, DA Lai, WANG Liwei, GAO Yuanshuai, ZHANG Wenjia, XIE Zhiqiang, YANG Huiguo, WANG Biao, LIU Yongbin. The Regulatory Role of Androgen Receptor in the Development of Ovarian Granulosa Cells in Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2026, 57(1): 246-257.

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基因名称 Gene name	引物序列（5′→3′） Primer sequences		退火温度/℃ Annealing temperature
AR	Forward	TATGGGGACATGCGTTTGGA	60
AR	Reverse	TGCTGGCACACAGGTACTTC	60
GAPDH	Forward	GTTTGTGATGGGCGTGAACC	58
GAPDH	Reverse	GCGTGGACAGTGGTCATAAGT	58
BCL2	Forward	GGGGTCATGTGTGTGGAGAG	60
BCL2	Reverse	TGCAGCTCCACAAAGGCGTC	60
BAX	Forward	TTCCGACGGCAACTTCAACT	60
BAX	Reverse	CTGATCAACTCGGGCACCTT	60
Caspase3	Forward	TTCAGAGGGGACTGTTGCAG	60
Caspase3	Reverse	CAGTCCAGTTCTGTGCCTCG	60
ESR2	Forward	AGAAGATTCCGGGCTTCGTG	60
ESR2	Reverse	GAGCAAAGATGAGCTTGCCG	60
CYP19A1	Forward	TCGTCCTGGTCACCCTTCTG	60
CYP19A1	Reverse	CGGTCTCTGGTCTCGTCTGG	60

雄激素受体对绵羊卵巢颗粒细胞发育的调控作用

The Regulatory Role of Androgen Receptor in the Development of Ovarian Granulosa Cells in Sheep

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