Effectiveness of 6-nitrobenzo[d]thiazol-2-amine derivative (N2) in ameliorating ethanol-induced fatty liver through lipogenesis inhibition and anti-inflammatory expression in a zebrafish model.

This study examines the effectiveness of the 6-nitrobenzo[d]thiazol-2-amine derivative (N2) in mitigating the effects of ethanol-induced fatty liver disease in zebrafish by assessing its anti-inflammatory, antioxidant, and antihyperlipidemic properties. The investigation encompasses the evaluation of N2 derivatives on hemolysis, cytotoxicity, developmental toxicity, oxidative stress, and lipid metabolism within zebrafish models. Antioxidant assays highlighted N2's significant radical scavenging activity. Cytotoxic evaluations conducted on HepG2 cells and human red blood cells (RBCs) revealed minimal cytotoxic effects and no hemolysis at concentrations up to 100 μM. Zebrafish embryo toxicity tests indicated slight developmental abnormalities at elevated concentrations (150 μM). Followed by reactive oxygen species (ROS) were assessed by DCFHDA staining in HepG2 cells. In zebrafish models subjected to 0.2 % ethanol exposure for 21 days, N2 treatment resulted in the elevation of antioxidant enzymes (superoxide dismutase, catalase), a reduction in lactate dehydrogenase levels, and an increase in glutathione levels, thereby suggesting a protective role against oxidative stress. Real-time PCR analysis demonstrated that N2 influences genes associated with lipid metabolism and inflammation. Furthermore, N2 substantially reduced ethanol-triggered elevations in reactive oxygen species, apoptosis, total cholesterol, and triglycerides within zebrafish livers. Histological analyses employing Oil Red O, Hematoxylin and Eosin (H&E), and Nile Red staining confirmed N2's ability to diminish hepatic steatosis and intracellular lipid accumulation. HPLC-UV further validated N2's efficacy in reducing lipid accumulation in zebrafish models with ethanol-induced fatty liver disease.