Chlorogenic acid attenuates malondialdehyde levels and regulates major ferroptosis-related genes in testes of acrylamide-exposed rats

Abstract

Acrylamide is a processing contaminant formed during high-temperature cooking of carbohydrate-rich foods and has been associated with impaired male reproductive function. This study aimed to determine the effects of chlorogenic acid on testicular lipid peroxidation and ferroptosis-related gene expression (GPX4, SLC7A11, and TFRC) in rats with acrylamide-induced testicular toxicity. Thirty-two male Wistar albino rats were randomized into four groups (n=8 per group): control, acrylamide-treated (ACR), chlorogenic acid–treated (CGA), and acrylamide + chlorogenic acid (ACR+CGA). Treatments were administered once daily by oral gavage for 28 days. At the end of the experimental period, testicular tissues were analyzed for malondialdehyde as an index of lipid peroxidation and for RT-qPCR assessment of GPX4, SLC7A11, and TFRC. Acrylamide exposure significantly increased malondialdehyde levels compared with controls, indicating enhanced lipid peroxidation. Conversely, chlorogenic acid co-treatment partially attenuated this increase, restoring malondialdehyde levels to near-control values. Gene expression analysis revealed that GPX4 was significantly upregulated 2.17-fold in the CGA group, while TFRC was significantly upregulated in both the CGA (2.66-fold) and ACR+CGA (2.34-fold) groups relative to the control, suggesting a potential adaptive response within the ferroptosis regulatory network. In silico protein-protein interaction network and functional enrichment analyses suggested that the GPX4 and TFRC interaction network is enriched for pathways related to selenoprotein metabolism and cellular iron handling. These findings indicate that chlorogenic acid influences testicular oxidative status and ferroptosis-related gene responses under acrylamide exposure, supporting its potential relevance as a dietary component for acrylamide-associated reproductive oxidative stress.