Cold plasma promotes Sertoli cell proliferation via AMPK mTOR signaling pathway

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) AMPK alpha 1 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.