Unsweetened Theobroma cacaopowder mitigates oxidative stress and preserves hepatic function in ethanol-fed Sprague Dawley rats

Background: Morphological evidence suggests that unsweetened cocoa powder (UCP) prevents hepatic injury in experimental ethanol-ingesting animal models, possibly due to its antioxidant and anti-inflammatory potentials. However, the functional integrity of hepatocytes and the underlying mechanisms are underexplored. Objective: This study investigated the protective effect of UCP on liver function in ethanol-ingesting male Sprague-Dawley rats. Methods: Male rats (200-220 g) were fed daily for 8 weeks with either 30% ethanol, 300 mg/kg UCP p.o, or 30% ethanol and 300 mg/kg UCP p.o. 6 h later. Rats that were neither fed with UCP nor ethanol served as controls. At the experimental endpoint, cardiac blood was collected for liver function test, prothrombin time and C-reactive protein assay. The large lobe of the liver was processed and stained with periodic acid Schiff (PAS) and lipofuschin. PAS-stained livers were used for hepatocyte glycogen estimation. The remaining lobes were homogenised for total antioxidant capacity and catalase activity assays. Results: Compared to the control, ethanol-only-fed rats showed lower serum total protein, albumin, and globulin, and lower hepatocyte glycogen, and higher prothrombin time. Also, ethanol-only-fed rats showed increased serum AST/ALT ratio, CRP, total bilirubin, and unconjugated bilirubin, but reduced conjugated bilirubin. Additionally, the livers of the ethanol-only-fed rats exhibited increased TAC, low catalase and increased lipofuscin accumulation. However, compared with ethanol-only-fed rats, UCP-treated ethanol-fed rats showed higher serum total protein, albumin, and globulin, and higher hepatocyte glycogen, and a lower prothrombin time. UCP-treated ethanol-fed rats also exhibited higher serum conjugated bilirubin, lower unconjugated bilirubin, less lipofuscin accumulation, and higher TAC and catalase levels compared to the ethanol-only-fed rats. Conclusion: Ingestion of UCP alongside ethanol not only attenuates hepatocellular damage but also preserves hepatic function, possibly by mitigating inflammation and oxidative stress. Thus, UCP could protect against alcohol-induced liver damage.