AMPK/SIRT1介导脂联素促进牦牛睾丸支持细胞乳酸转运的分子机制

doi:10.11843/j.issn.0366-6964.2025.07.041

摘要/Abstract

摘要：

通路在睾丸乳酸转运过程中发挥重要调控作用，且近年来脂联素对动物生殖的调控功能也逐渐被证明，但AMPK/SIRT1通路如何介导脂联素调控牦牛睾丸支持细胞的乳酸转运尚不明确。采集不同年龄牦牛睾丸(初生、幼年、成年及老年)各3头份，通过Western blot、组织免疫荧光染色、细胞免疫荧光染色等实验技术，探究AMPK/SIRT1通路介导脂联素促进牦牛睾丸支持细胞乳酸转运的分子机制。结果表明，AMPK、P-AMPK及SIRT1蛋白在不同年龄牦牛睾丸组织中均有表达，在幼年和老年时期的蛋白相对表达量极显著高于初生和成年(P < 0.01)。且AMPK及SIRT1蛋白在不同年龄牦牛睾丸中分布相似，主要分布在睾丸支持细胞、间质细胞及各级生精细胞。用不同浓度脂联素(0、5、10、20、50、100 ng ·mL^-1)处理牦牛睾丸支持细胞，当脂联素浓度为10 ng ·mL^-1时，极显著激活APNR1、AMPK、P-AMPK及SIRT1的蛋白表达(P < 0.01)，同时MCT1蛋白表达量最高，并且乳酸释放量最大。添加Compound C(AMPK蛋白抑制剂)能极显著抑制AMPK的蛋白表达(P < 0.01)，但脂联素和Compound C共处理能极显著提高AMPK、P-AMPK及MCT1蛋白表达量(P < 0.01)。结果提示，脂联素主要通过结合APNR1，激活AMPK/SIRT1信号通路促进牦牛睾丸支持细胞乳酸转运，在牦牛睾丸发育和精子发生中发挥作用。本研究为促进牦牛生产繁殖提供一定的理论基础和依据。

关键词: 牦牛, 睾丸支持细胞, 脂联素, 乳酸

Abstract:

The AMPK/SIRT1 pathway plays an important role in the regulation of testicular lactate transport, and the regulatory function of adiponectin on animal reproduction has been gradually proved in recent years. However, how the AMPK/SIRT1 pathway mediates adiponectin to regulate lactate transport in yak sertoli cells is still unclear. Three yak testis samples of different ages (newborn, juvenile, adult and aged) were collected. Western blot, tissue immunofluorescence staining and cell immunofluorescence staining were used to explore the molecular mechanism of the AMPK/SIRT1 pathway mediating adiponectin to promote lactate transport in sertoli cells of yak. Results showed that AMPK, P-AMPK and SIRT1 proteins were expressed in the testis tissue of yaks at different ages, and the relative protein expression levels in the juvenile and aged were significantly higher than those in the newborn and adults (P < 0.01). The distribution of AMPK and SIRT1 proteins was similar in the testis of yaks at different ages, mainly distributed in sertoli cells, leydig cells and spermatogenic cells at all levels. Sertoli cells were treated with various concentrations of adiponectin (0, 5, 10, 20, 50, 100 ng ·mL^-1). When the concentration of adiponectin was 10 ng ·mL^-1, The protein expression of APNR1, AMPK, P-AMPK and SIRT1 was significantly activated (P < 0.01), while the protein expression of MCT1 was the highest, and the lactate release was the highest. The protein expression of AMPK was significantly inhibited by Compound C (P < 0.01), an AMPK protein inhibitor. But the protein expression levels of AMPK, P-AMPK and MCT1 were significantly increased when adiponectin and Compound C were treated together (P < 0.01). The results indicate that APN plays a role in the development of the yak testis binding with APNR1. This study provides some theoretical basis for promoting yak production and reproduction.

Key words: yak, sertoli cell, adiponectin, lactate

中图分类号:

S852.1

张虔, 马睿, 崔燕, 余四九. AMPK/SIRT1介导脂联素促进牦牛睾丸支持细胞乳酸转运的分子机制[J]. 畜牧兽医学报, 2025, 56(7): 3495-3506.

ZHANG Qian, MA Rui, CUI Yan, YU Sijiu. Molecular Mechanism of AMPK/SIRT1 Mediating Adiponectin Promoting Lactate Transport in Yak Sertoli Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3495-3506.

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