Chasteberry extract alleviates rotenone-triggered testicular injury in rats by modulating oxidative stress, inflammation, apoptosis, and mitochondrial dysfunction

Widespread usage of the pesticide rotenone has been linked to several health issues in both people and animals. Thus, we aimed to counteract rotenone-induced testicular oxidative damage using Chasteberry extract (CBE), while concurrently investigating the underlying mechanisms. Forty male albino rats were randomized into four groups (10 rats per each group): served as control (received 1 mL of saline orally, and 2% dimethyl sulfoxide (DMSO) at a dose of 10 mL/kg for 3 times/week, intraperitoneally), CBE (165 mg/kg/daily, orally), rotenone (3 mg/kg/3times/week, intraperitonially), and combination (CBE and rotenone). Treatments were administered for 60 consecutive days. Our data indicated that rotenone injection resulted in hormonal imbalance, oxidative damage, inflammation, apoptosis, as well as mitochondrial dynamic dysfunction in testicular tissue. While the co-administration of CBE with rotenone significantly mitigated these deleterious effects and restored sperm morphology and count, as evidenced biochemically by upregulation in follicle-stimulating hormone, luteinizing hormone and testosterone hormonal levels. Also, the CBE-supplemented rotenone group showed enhanced antioxidant enzyme activities (superoxide dismutase, catalase, total and reduced glutathione) and reduced malondialdehyde levels, oxidized glutathione, and proinflammatory cytokine levels (tumor necrosis factor and interleukin 1β). These results were further confirmed by histopathological investigation, which revealed marked restoration of testicular histoarchitecture with reduction in nuclear factor kappa B and caspase 3 immunoreactivity in the combination group. On the molecular level, CBE supplementation restored the antioxidant mRNA transcript of antioxidant genes, mitochondrial biogenesis genes, and mitochondrial dynamics, as evidenced by upregulation in mitofusin and downregulation in dynamin-related protein 1. In conclusion, CBE is a promising protective agent against rotenone-induced testicular damage via its antioxidant, anti-inflammatory, antiapoptotic, and mitochondrial biogenesis-enhancing effects.