Evaluation of the Effect of Copper Sulfate Exposure on Organs in Juvenile Rats

Copper sulphate pentahydrate is widely used in agriculture to control bacterial and fungal diseases in various crops. Despite its extensive application, limited data exist regarding its potential toxic effects on juvenile rats following early-life exposure. In addition to oxidative stress and inflammation, copper overload may also trigger cuproptosis, a recently identified copper-dependent form of regulated cell death. This study aimed to investigate the histopathological, biochemical, and molecular effects of copper sulphate exposure on major organs in juvenile rats and to elucidate the associated inflammatory and oxidative stress-related mechanisms. Male and female Sprague–Dawley rats (30–40 days old, 50–70 g) were randomly assigned to control and experimental groups. Following copper sulphate exposure, histopathological examinations were performed on major organs, including the liver, kidney, heart, lung, and reproductive tissues (testis in males and ovary in females). Immunohistochemical analyses of tumor necrosis factor-alpha (TNF-α) and nuclear factor kappa B (NF-κB) were conducted. Oxidative stress parameters, including malondialdehyde (MDA), total antioxidant status (TAS), and total oxidant status (TOS), were measured using ELISA. Gene expression levels of TNF-α and NF-κB were evaluated by quantitative real-time PCR (qRT-PCR). Copper sulphate exposure induced significant histopathological alterations in all examined tissues of both male and female juvenile rats. Biochemical findings revealed increased oxidative stress, evidenced by elevated MDA and TOS levels along with altered TAS values. Furthermore, immunohistochemical and gene expression analyses demonstrated upregulation of TNF-α and NF-κB, indicating activation of inflammatory pathways. Copper sulphate exposure leads to widespread morphological changes in juvenile rats, potentially mediated by oxidative stress and inflammation. These findings provide insight into the biological impact of early-life pesticide exposure. Further studies are warranted to clarify the underlying molecular mechanisms and to develop effective preventive or therapeutic approaches.