PHYTOCHEMICAL COMBINATION THERAPY FOR LIVER FIBROSIS: INVESTIGATING APOCYNIN AND URSOLIC ACID IN A CCL₄ INDUCED RAT MODEL

Abstract

Background: Hepatic fibrosis is characterized by excessive accumulation of extracellular matrix, leading to progressive disruption of liver function. Activation of hepatic stellate cells (HSCs), often driven by oxidative stress, plays a central role in fibrogenesis. Apocynin and Ursolic acid are plant-derived compounds with reported antioxidant and antifibrotic properties. This study aimed to evaluate their individual and combined therapeutic effects in a carbon tetrachloride (CCl₄)-induced hepatic fibrosis rat model.

Objective: To investigate the potential synergistic antifibrotic effects of Apocynin and Ursolic acid in CCl₄-induced hepatic fibrosis.

Method: Thirty Wistar rats were divided into five groups: normal control, disease control (CCl₄), and three treatment groups receiving Apocynin (3 mg/kg), Ursolic acid (20 mg/kg), or their combination. Hepatic fibrosis was induced with CCl₄ (1 ml/kg, intraperitoneally, twice weekly for 3 weeks), followed by a 21-day oral treatment period. Biochemical parameters including SGPT, SGOT, ALP, GGT, hydroxyproline, total protein, and albumin were evaluated. Oxidative stress markers such as reduced glutathione, catalase activity, TBARS, and tissue hydroxyproline were measured. Histopathological analysis was performed using H&E and Masson’s trichrome staining.

Results: Both Apocynin and Ursolic acid significantly reduced serum and tissue hydroxyproline levels and improved antioxidant enzyme activity. Histological analysis indicated attenuation of fibrosis in treated groups. However, no significant synergistic enhancement was observed with the combination therapy.

Conclusion: Apocynin and Ursolic acid exhibit promising antifibrotic and antioxidant activity in a CCl₄-induced hepatic fibrosis model. Further studies with prolonged treatment durations are warranted to explore potential synergistic effects.