Development and evaluation of chitosan conjugated Isatin-linked Pyrazole derivative to target anti-inflammatory and anti-fibrotic in ethanol induced liver fibrosis using an in-vivo zebra fish model.

Alcoholic liver disease (ALD) remains a health burden, characterized by hepatic steatosis to fibrosis, a significant contributor to global morbidity and mortality, with limited therapeutic options for advanced stages like liver fibrosis. This study explores the antifibrotic and anti-inflammatory potential of a novel isatin-linked pyrazole derivative (3F) conjugated with chitosan-EDTA (CS) in mitigating ethanol (EtOH) induced liver fibrosis in zebrafish model. We demonstrated hepatic fibrosis using a chronic low-dose EtOH model (0.2 %), mimicking the ALD stage. CS-3F conjugates were synthesized using ionotropic gelation, exhibiting optimal drug-loading capacity, sustained drug release, and enhanced bioavailability. Morphological characterization revealed particle sizes ranging from 50 to 300 nm, facilitating efficient cellular uptake and liver-targeted drug distribution. EtOH exposure significantly elevated hepatic biomarkers (ALT, AST, and ALP), disrupted lipid metabolism (TC and TG), and impaired detoxification functions (ammonia and urea metabolism). Oxidative stress and inflammation were evident, with decreased SOD and CAT levels, increased MDA and LDH levels, and upregulation of pro-inflammatory and fibrotic genes (il-1β, tnfα, tgf-β, col1a1, and fasn). Treatment with CS-3F significantly ameliorated these alterations, restoring metabolic, detoxification, antioxidant functions and significantly reducing inflammation and fibrosis. Histopathological analysis confirmed improvements in liver architecture, including reduced parenchymal damage and ECM deposition. These results highlight the utility of CS-3F in reversing fibrosis and restoring liver function, offering a promising avenue for addressing the unmet therapeutic needs in ALD. However, further studies are needed to validate these findings in mammalian models and elucidate the underlying molecular mechanisms.