Journal of Integrative Agriculture ›› 2022, Vol. 21 ›› Issue (9): 2700-2719.DOI: 10.1016/j.jia.2022.07.010

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JIA-2021-0512 低温等离子体通过AMPK-mTOR信号通路促进睾丸支持细胞的增殖

  

  • 收稿日期:2021-04-16 接受日期:2022-01-04 出版日期:2022-09-01 发布日期:2022-01-04

Cold plasma promotes Sertoli cell proliferation via AMPK-mTOR signaling pathway

ZHANG Jiao-jiao, LI Ya-qi, SHI Mei, WANG Yu-sha, TANG Yao, WANG Xian-zhong   

  1. Chongqing Key Laboratory of Forage and Herbivore, College of Veterinary Medicine, Southwest University, Chongqing 400715, P.R.China

  • Received:2021-04-16 Accepted:2022-01-04 Online:2022-09-01 Published:2022-01-04
  • About author:ZHANG Jiao-jiao, E-mail: zhangjjff@126.com; Correspondence WANG Xian-zhong, E-mail: xianzhong_wang@aliyun.com
  • Supported by:
    This research was supported by the National Natural Science Foundation of China (31902338), the Natural Science Foundation of Chongqing, China (cstc2019jcyj-msxmX0056), and the Innovative Project of Chongqing Returned Overseas Person Entrepreneurship and Innovation Plan, China (cx2020057).

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摘要：

本研究旨在探讨低温等离子体对仔鸡睾丸支持细胞增殖的影响及其调控机制。结果发现，用2.4 W放电功率的低温等离子体间隔6 h处理两次，每次处理时间30 s，对支持细胞的活性、生长速度和细胞的周期进程具有最大的促进作用（P<0.05）。低温等离子体处理增加了睾丸支持细胞线粒体的活性、三磷酸腺苷的产生和呼吸链酶的活性（P<0.05），减少了细胞内活性氧的生成（P<0.05），提高了抗氧化酶的活性（P<0.05），增加了miR-7450-5p的表达（P<0.05），使腺苷一磷酸活化蛋白激酶（adenosine monophosphate-activated protein kinase, AMPK）的水平降低（P<0.05），并且降低了miR-100-5p的表达（P<0.05），使哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin, mTOR）的水平升高（P<0.05）。支持细胞转染miR-7450-5p抑制剂降低了miR-7450-5p的表达（P<0.05），增加了AMPK的水平（P<0.05），转染miR-100-5p模拟物增加了细胞miR-100-5p的表达（P<0.05），降低了mTOR的水平（P<0.05）。转染miR-7450-5p抑制剂和miR-100-5p模拟物均显著降低了睾丸支持细胞的活性和生长（P<0.05），抑制了细胞周期进程（P<0.05），减少了线粒体的活性、三磷酸腺苷的水平和呼吸链酶的活性（P<0.05），然而低温等离子体处理可以显著改善miR-7450-5p抑制剂和miR-100-5p模拟物对支持细胞增殖的抑制作用（P<0.05）。研究结果表明，低温等离子体处理可能通过影响miRNAs水平与活性氧稳态来调控AMPK-mTOR信号通路，进而通过增加线粒体三磷酸腺苷水平和呼吸链酶活性促进睾丸支持细胞的增殖。本研究优化了低温等离子体促进睾丸支持细胞增殖的处理条件，阐述了其调控的可能机制，为临床实际中利用低温等离子体技术促进睾丸支持细胞增殖奠定基础，从而有利于提高公鸡的繁殖性能。


Abstract:

This study investigated cold plasmas for multiple biological applications.  Our previous work has found dielectric barrier discharge plasma improves chicken sperm quality.  The number of Sertoli cells (SCs) decides spermatogenesis.  However, whether cold plasma can regulate SC proliferation remains unclear.  This study explored the effects of cold plasma on immature chicken SC proliferation and the regulation mechanism.  Results showed that cold plasma exposure at 2.4 W for 30 s twice with an interval of 6 h produced (P<0.05) the maximum SC viability, cell growth, and cell cycle progression.  SC proliferation-promoting effect of cold plasma treatment was regulated by increasing (P<0.05) the adenosine triphosphate production and the respiratory enzyme activity in the mitochondria.  This process was potentially mediated by the adenosine monophosphate-activated protein kinase (AMPK)–mammalian target of rapamycin (mTOR) signaling pathway, which was regulated by the microRNA (miRNA) targeting regulation directly and by the intracellular reactive oxygen species homeostasis indirectly.  The cold plasma treatment increased (P<0.01) the miR-7450-5p expression and led to a decreased (P<0.01) AMPKα1 level.  On the other hand, miR-100-5p expression was reduced (P<0.05) and led to an increased (P<0.05) mTOR level in SCs.  A single-stranded synthetic miR-7450-5p antagomir and a double-stranded synthetic miR-100-5p agomir reduced (P<0.05) the SC proliferation.  However, this could be ameliorated (P<0.05) by the cold plasma treatment.  Our findings suggest that appropriate cold plasma treatment provides a safe strategy to improve SC proliferation, which is beneficial to elevating male chicken reproductive capacity.

Key words: cold plasma , Sertoli cell proliferation , AMPK , mTOR , miRNAs