Title : Synergistic antifibrotic potential of protocatechuic acid and D-carvone in liver protection
Abstract:
Background: Liver fibrosis, driven by chronic injury, oxidative stress, and inflammation, represents a significant global health burden. Natural compounds such as protocatechuic acid (PCA) and D-Carvone (D-Car) possess known antioxidant and anti-inflammatory properties, but their combined therapeutic potential against liver fibrosis remains unexplored.
Purpose: This study aimed to investigate the synergistic antifibrotic effects of PCA and D-Car in both cellular and animal models of liver fibrosis.
Methods: A combined in vitro and in vivo approach was employed. In vitro experiments utilized the HSC-T6 hepatic stellate cell line, where cells were treated with PCA (50 μM) and D-Car (50 ng/mL), individually and in combination. Cytotoxicity (MTT assay), apoptosis (flow cytometry and Hoechst staining), and Annexin A2 levels were assessed. In vivo studies involved a CCl₄-induced liver fibrosis rat model. Following treatment with PCA, D-Car, or their combination, body weight, liver enzymes (ALT, AST, ALP, albumin), LDH activity, fibrotic markers (TIMP-1, Col1α1), antioxidant enzymes (SOD, Cyp2e1), inflammatory cytokines (IL-6, TNF-α, NF-κB), and histological changes were evaluated.
Results: PCA+D-Car treatment significantly improved HSC-T6 cell morphology, reduced cytotoxicity (*p* < 0.001), and decreased apoptosis compared to individual treatments. In vivo, PCA+D-Car restored body weight, normalized liver enzymes, downregulated TIMP-1 and Col1α1, enhanced antioxidant activity (SOD1), and reduced inflammatory markers (*p* < 0.01). Histological analysis confirmed improved liver architecture with reduced fibrosis and inflammation.
Conclusion: The combination of PCA and D-Carvone exhibits synergistic antifibrotic effects by mitigating oxidative stress, inflammation, and extracellular matrix deposition. These findings highlight its potential as a dietary or therapeutic strategy for liver fibrosis, warranting further clinical exploration.
