Title : Synergistic antifibrotic potential of protocatechuic acid and D-Carvone in liver protection
Abstract:
Liver fibrosis represents a critical global health burden with limited therapeutic options. This study investigates the novel synergistic efficacy of two natural compounds—Protocatechuic Acid (PCA), a phenolic acid, and D-Carvone (D-Car), a monoterpene—in mitigating liver fibrosis using integrated in vitro and in vivo approaches.
In the in vitro arm, the HSC-T6 hepatic stellate cell line was treated with PCA (50 μM) and D-Car (50 ng/mL), both individually and in combination. The PCA+D-Car group demonstrated significant restoration of cell morphology, reduction in cytotoxicity (MTT assay, p<0.001), and suppression of apoptosis (flow cytometry and Annexin A2 ELISA), outperforming monotherapies and approximating the profile of normal liver cells.
For in vivo validation, a CCl?-induced liver fibrosis model in Sprague-Dawley rats (n=6 per group) was employed. Co-administration of PCA (4 mg/kg) and D-Car (50 mg/kg) markedly improved body weight recovery (36.41% vs. 25.42% in CCl? group) and liver function, indicated by normalized levels of ALT, AST, ALP, and albumin (p<0.001). The combination therapy also significantly reduced lactate dehydrogenase (LDH), downregulated fibrogenic genes (TIMP-1 and Col1α1), enhanced antioxidant defense (SOD1 upregulation, Cyp2e1 and iNOS suppression), and attenuated pro-inflammatory cytokines (IL-6, TNF-α, NF-κB). Histopathological analysis confirmed notable architectural restoration and reduced collagen deposition.
These results underscore a powerful synergism between PCA and D-Car in counteracting fibrogenesis, oxidative injury, and inflammation—key drivers of liver pathology. The combination targets multiple signaling hubs, offering a multifaceted therapeutic strategy derived from natural sources. Our findings position PCA+D-Car as a promising candidate for dietary intervention or adjunctive therapy in hepatic disorders, warranting further clinical exploration into its translational potential.
