Biosynthesis of silver nanoparticles using Pterorhachis zenkeri: Characterization and evaluation of antioxidant, anti-apoptotic, and androgenic properties in TM3 Leydig cells exposed to cyclophosphamide

Abstract

We fabricated silver nanoparticles (AgNPs) using Pterorhachis zenkeri, characterized by Atomic Force Microscopy (AFM) and determined their antioxidant potentials in vitro. Results confirmed the fabrication of AgNPs by using P. zenkeri as a bioreducing agent for the first time. AgNPs possessed potent antioxidant activity in vitro. Furthermore, the TM3 cells were treated for 24 h with AgNPs, vitamin E and cyclophosphamide (CP) at different concentrations (25, 50, 100, 250 and 500 µg/ml). The cells morphology, apoptosis, mitochondrial membrane depolarisation (MMD), Reactive Oxygen Species (ROS), caspase 3/9, oxidative stress-related enzymes, testosterone, and the mRNAexpression of steroidogenic acute regulatory protein (StAR) were measured. Results revealed that AgNPs, vitamin E and CP decreased the cell viability in a dose-dependent manner, but did not affect the TM3 cells morphology after treatment. The cytotoxicity of CP in TM3 cells was alleviated after AgNPs application. For instance, AgNPs significantly (p<0.001-0.05) reduced the MMD, ROS production, and caspase 3/9 activities, but increased the activities of superoxide dismutase, catalase and glutathione peroxidase in the TM3 cells. Moreover, AgNPs improved testosterone production by activating StAR machineries. These results indicate that AgNPs/P. zenkeri could be a potential alternative drug in the management of oxidative stress and androgen deficit associated with CP chemotherapy.