Abstract
Non-thermal plasma treatment is an emerging innovative technique with a wide range of biological applications. This study was conducted to investigate the effect of a non-thermal dielectric barrier discharge plasma technique on immature chicken Sertoli cell (SC) viability and the regulatory role of microRNA (miR)-7450. Results showed that plasma treatment increased SC apoptosis in a time- and dose-dependent manner. Plasma-induced SC apoptosis possibly resulted from the excess production of reactive oxygen species via the suppression of antioxidant defense systems and decreased cellular energy metabolism through the inhibition of adenosine triphosphate (ATP) release and respiratory enzyme activity in the mitochondria. In addition, plasma treatment downregulated miR-7450 expression and activated adenosine monophosphate-activated protein kinase α (AMPKα), which further inhibited mammalian target of rapamycin (mTOR) phosphorylation in SCs. A single-stranded synthetic miR-7450 antagomir disrupted mitochondrial membrane potential and decreased ATP level and mTOR phosphorylation by targeting the activation of AMPKα, which resulted in significant increases in SC lethality. A double-stranded synthetic miR-7450 agomir produced opposite effects on these parameters and ameliorated plasma-mediated apoptotic effects on SCs. Our findings suggest that miR-7450 is involved in the regulation of plasma-induced SC apoptosis through the activation of AMPKα and the further inhibition of mTOR signaling pathway.
Highlights
The use of non-thermal plasma is receiving great interest in various biomedical applications, including sterilization, blood coagulation, wound healing, tissue regeneration, dental treatment, promotion of cell transfection efficiency, cell proliferation and differentiation, and cancer therapy[1,2]
We investigated the molecular and cellular mechanisms of non-thermal dielectric barrier discharges (DBDs) plasma-induced in vitro effects on chicken Sertoli cell (SC), with particular emphasis on the roles of intracellular reactive oxygen species (ROS) and energy metabolism in regulating gene expression and cellular signaling pathways, which may provide a potential reference for the vivo application in future
Green fluorescence of JC-1 dye indicates a decrease in mitochondrial membrane potential, which is observed early in cell apoptosis, and mitochondrial depolarization occurring during apoptosis is indicated by an increase in the green/red fluorescence intensity ratio
Summary
The use of non-thermal plasma is receiving great interest in various biomedical applications, including sterilization, blood coagulation, wound healing, tissue regeneration, dental treatment, promotion of cell transfection efficiency, cell proliferation and differentiation, and cancer therapy[1,2]. The present study was carried out with the objectives to investigate the effect of non-thermal plasma treatment on immature chicken Sertoli cell (SC) viability and growth in vitro and the exploration of plasma exposure condition before its in vivo application. We investigated the molecular and cellular mechanisms of non-thermal DBD plasma-induced in vitro effects on chicken SCs, with particular emphasis on the roles of intracellular ROS and energy metabolism in regulating gene expression and cellular signaling pathways, which may provide a potential reference for the vivo application in future. Ablation of SC dicer, which is an enzyme essential for SC maturation and survival and is required to sustain germ cell development, leads to infertility This observation indicates that miRNA expression in SCs is important in supporting germ cell development[32]. The present study aimed to elucidate the role of miR-7450 in the regulation of non-thermal plasma-mediated effects on SCs through the AMPK-mTOR signaling pathway
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