| 摘要: |
| 【目的】探究转化生长因子-β1（TGF-β1）在小尾寒羊卵巢颗粒细胞黄体化过程中的作用，为解析TGF-β1调控小尾寒羊卵巢颗粒细胞黄体化的机制提供理论依据。【方法】对小尾寒羊母羊进行同期发情处理，采用HE染色、RT-qPCR、Western blot 和免疫荧光等方法分析TGF-β1在成熟卵泡、红体及黄体中的表达水平。体外培养小尾寒羊卵巢颗粒细胞，在培养基中分别添加5和10 IU/mL人绒毛膜促性腺激素（hCG）及1和2 μg/mL胰岛素，筛选颗粒细胞体外黄体化的适宜培养基。在适宜培养基的基础上添加10 ng/mL TGF-β1重组蛋白，研究其对小尾寒羊卵巢颗粒细胞黄体化进程的影响。【结果】TGF-β1在小尾寒羊卵巢黄体组织，特别是颗粒黄体细胞中高表达（P<0.01）。2 μg/mL胰岛素为诱导颗粒细胞黄体化的适宜条件，能引起颗粒细胞超微结构的变化，促进黄体化相关基因（StAR、HSD3B1）、类固醇激素（E2和P4）及其合成限制酶（3β?HSD、17β?HSD、P450scc）的表达（P<0.05），显著促进细 胞活力、增殖，抑制细胞凋亡。TGF-β1能显著上调黄体化相关基因StAR和HSD3B1的表达，并促进类固醇激素（E2和P4）的合成（P<0.05）。【结论】TGF-β1可促进小尾寒羊卵巢颗粒细胞黄体化。 |
| 关键词: 转化生长因子-β1 小尾寒羊 颗粒细胞黄体化 胰岛素 人绒毛膜促性腺激素 |
| DOI：10. 13207/j. jnwafu. 2027. 01. 008 |
| 分类号: |
| 基金项目:甘肃省高校教师创新基金项目（2025A-083）；甘肃农业大学科技创新基金-青年导师扶持基金项目（GAU-QDFC-2024-04）；甘肃省畜牧技术推广总站委托项目（GSAU-JSKF-2025-18）；浙江大学动物分子营养学教育部重点实验室开放基金项目（KLMAN202502） |
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| Biological function of transforming growth factor⁃β1 during luteinization of ovarian granulosa cells in Small⁃Tailed Han sheep |
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Zhao Zhihui, Ren Siyu, Cui Jingjing, Guo Yajing, Cui Mengru, Wang Xiaolong, Li Mingna, Wang Jiqing, Hao Zhiyun
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College of Animal Science and Technology，Gansu Agricultural University，Lanzhou，Gansu 730070，China
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| Abstract: |
| 【Objective】This study aims to investigate the role of TGF- β1 during luteinization of ovarian granulosa cells in Small-Tailed Han sheep，providing a theoretical reference for elucidating the regulatory mecha? nism of TGF- β1 in the process.【Method】This study performed estrus synchronization treatment on SmallTailed Han sheep ewes，and employed methods including HE staining，RT-qPCR，Western blot，and immuno? fluorescence to analyze the expression levels of TGF- β1 in mature follicles，corpora lutea，and corpora rubra. Ovarian granulosa cells were cultured in vitro and treated with 5 and 10 IU/mL hCG and 1 and 2 μg/mL insulin to screen the optimal medium for in vitro luteinization. Then，10 ng/mL TGF- β1 recombinant protein was added to the optimal medium to determine its effects on granulosa cell luteinization.【Result】The results showed that TGF-β1 was highly expressed in the ovarian luteal tissue of Small-Tailed Han sheep，particularly in granulosa lutein cells（P<0.01）.2 μg/mL insulin was identified as an optimal condition for inducing granulosa cell luteinization，which induced ultrastructural changes，promoted the expression of luteinization-related genes （StAR，HSD3B1），steroid hormones （E2 and P4），and their rate-limiting enzymes （3β -HSD，17β -HSD，P450scc）（P<0.05），and significantly enhanced cell viability and proliferation while inhibiting apoptosis. TGF-β1
significantly upregulated the expression of luteinization-related genes StAR and HSD3B1，and promoted the synthesis of steroid hormones（E2 and P4）（P<0.05）.【Conclusion】TGF-β1 can promote luteinization of ova-rian granulosa cells in Small-Tailed Han sheep. |
| Key words: transforming growth factor-β1 Small-Tailed Han sheep granular cell luteinization insulin human chorionic gonadotropin |
