Comprehensive Study on the Hepatoprotective Effects of Chlorogenic Acid and Vitamin E in a Mouse Model of Diclofenac-Induced Injury

Abstract

Background: Diclofenac (DIC) is a commonly used non-steroidal anti-inflammatory drug (NSAID) associated with hepatotoxicity through oxidative stress and inflammatory pathways. Objectives: This study aimed to investigate the protective effects of chlorogenic acid (CGA) in both preventive and therapeutic regimens compared with vitamin E against DIC-induced liver injury in mice. Methods: This interventional, experimental, randomized study was conducted using a mouse model. Propylene glycol (PPG) was used as the solvent for CGA. Forty-two BALB/C mice were randomly assigned into seven groups: Control, CGA, DIC, PPG+DIC, CGA+DIC (therapeutic regimen), vitamin E+DIC, and CGA+DIC(P) (preventive regimen). Treatments were administered for 7 consecutive days. Serum liver enzymes, total bilirubin, hepatic oxidative stress markers, and inflammatory gene expression were measured. Liver samples were also collected for histopathological evaluation. Data were analyzed using ANOVA followed by Tukey’s post hoc test, with statistical significance set at P < 0.05. Results: Diclofenac significantly increased the expression of tumor necrosis factor (TNF-α) and interleukin-1 beta (IL-1β) (P < 0.05). It also elevated malondialdehyde (MDA) levels and reduced the activities of antioxidant enzymes, including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) (P < 0.05). Furthermore, DIC markedly increased serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), and total bilirubin (P < 0.05). Chlorogenic acid alone showed no significant changes compared to the control group (P > 0.05). Both preventive and therapeutic regimens of CGA significantly attenuated DIC-induced upregulation of TNF-α and IL-1β and improved oxidative stress markers, with complete normalization of MDA and partial recovery of CAT and GPx (P < 0.05). Liver enzymes decreased significantly in all treated groups, with AST and GGT returning to control levels (P < 0.05). The effects of CGA were comparable to those of vitamin E in all measured parameters. Histopathology confirmed that CGA-treated groups exhibited milder hepatocellular degeneration than the DIC group, with effects similar to those of vitamin E. Conclusions: Chlorogenic acid exerts protective effects on the liver in DIC-induced injury by reducing oxidative stress (enhancing CAT, SOD, and GPx activities; lowering MDA), suppressing inflammation (IL‑1β and TNF‑α), and improving histopathological and biochemical liver markers (ALT, AST, ALP, GGT, and total bilirubin). These effects were observed in both preventive and therapeutic regimens and were comparable to those of vitamin E. These findings indicate the potential of CGA as a natural agent for mitigating drug-induced hepatotoxicity.