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。

Estrogen plays an important role in regulating testicular Sertoli cell number. Furthermore, S-phase kinase-associated protein 2 (SKP2) plays a central role in mammalian cell cycle progression. The objective of this study was to determine whether 17β-estradiol can regulate the expression of SKP2, and the Sertoli cell cycle, via estrogen receptor β (ERβ), the cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) and extracellular signal-regulated kinase (ERK1/2) pathway. When cultured immature boar Sertoli cells were treated with 17β-estradiol, a time-dependent increase in SKP2 mRNA and protein level was observed by real-time PCR and Western blot, and 17β-estradiol activity peaked at 30 min. Treatment with ICI182780 and ERβ antagonist reduced 17β-estradiol-induced expression of SKP2 and proliferating cell nuclear antigen (PCNA), while increasing the protein concentration of p27kip1. However, the effect of ERa antagonist on these parameters was lower than that of ICI182780 and ERβ. Forskolin had a similar effect as 17β-estradiol on the expression of SKP2, PCNA and p27kip1. Rp-cAMP, H-89 and U0126 treatment reduced 17β-estradiol-induced changes, while H-89 also inhibited ERK1/2 activation. Therefore, 17β-estradiol mainly regulates SKP2 mRNA and protein expression via ERβ-cAMP-PKA and ERK1/2 activation. SKP2 and PCNA expression were positively correlated, while increased SKP2 expression likely resulted in p27kip1 degradation.