Evaluation of the therapeutic potential of novel nanoparticle formulations of glutathione and virgin coconut oil in an experimental model of carbon tetrachloride-induced liver failure

Abstract

Background: Acute liver failure (ALF) is a critical condition characterized by rapid liver dysfunction, leading to high mortality rates. Current treatments are limited, primarily supportive, and often require liver transplantation. This study investigates the potential of a novel nanoparticle formulation of glutathione (GSH) and virgin coconut oil (VCO) alone and in combination to enhance therapeutic outcomes in a rat model of ALF induced by orogastric carbon tetrachloride (CCl₄).

Methods: The study employed adult male Albino rats divided into ten groups, with ALF induced via a single oral dose of CCl₄. Various treatment regimens were administered over seven days, including conventional and nanoparticle forms of GSH and VCO and their combinations. The efficacy of treatments was evaluated through biochemical analysis of liver function markers, oxidative stress indicators, inflammatory biomarkers, and histopathological examinations. Nanoparticles were synthesized using established methods, and characterization techniques were employed to ensure their quality and properties.

Results: The nanoparticle formulations significantly improved liver function, as indicated by reduced serum levels of alanine aminotransferase and aspartate aminotransferase, alongside decreased oxidative stress markers such as malondialdehyde. Furthermore, they reduced tumor necrosis factor alpha and interleukin-1 beta inflammatory markers. Histological analysis revealed reduced hepatocellular necrosis and inflammation in treated groups compared to controls. Also, decreased nuclear factor-kappa B was detected by immunohistochemical analysis.

Conclusion: The findings show that the nanoparticle mixture of GSH and VCO effectively reduces liver damage in ALF. This suggests a promising drug-based approach for improving liver regeneration and protection. This innovative strategy may pave the way for new therapeutic interventions in the management of ALF.

Keywords: ALF; Carbon tetrachloride; IL-1β; NF-κB; Nanoparticles; VCO.

Fig. 1

The impact of different forms of glutathione (GSH) and virgin coconut oil (VCO), including conventional, nanoparticle, and combination forms, on blood levels of aspartate aminotransferase (AST), alanine transaminase (ALT), albumin (ALB), and alkaline phosphatase in rats with acute liver failure induced by carbon tetrachloride (CCl₄). Data are the means ± SEM (n = 6). ^bp < 0.001 and ^ap < 0.0001 as compared with the control group. **p < 0.01 and ****p < 0.0001 as compared to the CCl₄-treated group. ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 and ^####p < 0.0001 when contrasted with the related nanoparticle category. Note FO ferric oxide, CS chitosan, Nano-VCO virgin coconut oil nanoparticles, Nano-GSH glutathione nanoparticles, O virgin coconut oil-glutathione combinations, Nano-O virgin coconut oil-glutathione nanoparticle combinations

Fig. 2

The impact of different forms of glutathione (GSH) and virgin coconut oil (VCO), including conventional, nanoparticle, and combination forms, on blood levels of bilirubin (direct and total) and gamma-glutamyl transferase in rats with acute liver failure induced by carbon tetrachloride (CCl₄). Data are the means ± SEM (n = 6). ^ap < 0.0001 as compared with the control group. **p < 0.01 and ****p < 0.0001 as compared to the CCl₄-treated group. ^#p < 0.05, ^##p < 0.01 and ^###p < 0.001 when contrasted with the related nanoparticle category. Note FO ferric oxide, CS chitosan, Nano-VCO virgin coconut oil nanoparticles, Nano-GSH glutathione nanoparticles, O virgin coconut oil-glutathione combinations, Nano-O virgin coconut oil-glutathione nanoparticle combinations

Fig. 3

The impact of different forms of glutathione (GSH) and virgin coconut oil (VCO), including conventional, nanoparticle, and combination forms, on tissue levels of interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in rats with acute liver failure induced by carbon tetrachloride (CCl₄). Data are the means ± SEM (n = 6). ^ap < 0.0001 as compared with the control group. ****p < 0.0001 as compared to the CCl₄-treated group. ^##p < 0.01 and ^####p < 0.0001 when contrasted with the related nanoparticle category. Note FO ferric oxide, CS chitosan, Nano-VCO virgin coconut oil nanoparticles, Nano-GSH glutathione nanoparticles, O virgin coconut oil-glutathione combinations, Nano-O virgin coconut oil-glutathione nanoparticle combinations

Fig. 4

The impact of different forms of glutathione (GSH) and virgin coconut oil (VCO), including conventional, nanoparticle, and combination forms, on tissue levels of malondialdehyde (MDA) and glutathione peroxidase (GPx) in rats with acute liver failure induced by carbon tetrachloride (CCl₄). Data are the means ± SEM (n = 6). ^ap < 0.0001 as compared with the control group. ****p < 0.0001 as compared to the CCl₄-treated group. ^#p < 0.05, ^##p < 0.01 and ^####p < 0.0001 when contrasted with the related nanoparticle category. Note FO ferric oxide, CS chitosan, Nano-VCO virgin coconut oil nanoparticles, Nano-GSH glutathione nanoparticles, O virgin coconut oil-glutathione combinations, Nano-O virgin coconut oil-glutathione nanoparticle combinations

Fig. 5

A representative image of liver tissue with HE staining A control group within normal structure of the liver lobules, central vein, and surrounding hepatocytes is clearly visible, B CCl₄ group show moderate to severe vacuolar degeneration (arrowhead) and multifocal necrosis (yellow arrow) with multifocal inflammatory cell infiltration (black arrow) and hemorrhage in between hepatocyte (star), C and D CS and FO groups show focal hepatic necrosis mixed with mononuclear inflammatory cells (arrow) surrounded with hepatocytes reveal marked vacuolation (arrowhead), E and F represent GSH and VCO show mild improvement of liver tissue within focal hepatic necrosis mixed with mononuclear inflammatory cells and diffuse moderate vacuolar changes within the hepatocytes (arrowhead) and presence of mitotic figure (circle), G, H Nano-CS and Nano-VCO show more improvement degree of hepatic tissue with presence of mitotic figure and mild infammatory cells infilterated, I represents conventional combination (O) of treatments show normal hepatic parenchyma with midzonal area of fatthy changes in hepatic parenchyma. J that represent nano-combination (Nano-O) treatments show more healthy hepatic tissue within mild degree of periportal hepatic vacuolation consistent with mild fatty change (arrowhead). Scale bar = 50 μm. CCl₄ carbon tetrachloride, CS chitosan, GSH glutathione, Fe₃O₄ ferric oxide, VCO virgin coconut oil

Fig. 6

Immunohistochemical analysis revealed A the absence of NF-κB expression in the liver tissue of the control group. B CCl₄ treatment significantly increased NF-κB expression in the cytoplasm and nucleus of hepatocytes, as indicated by brown staining (arrowhead). C and D CS and FO groups showing less increase in NF-κB immunostaining. E and F showing decrease of NFκB-P65 antibody within the hepatic cells in treated groups with GSH, and VCO. G, H treatment groups with Nano-GSH and Nano-VCO showed an obvious decrease in both cytoplasmic and nuclear expression of the NFκB-P65 antibody (arrowhead). I The combined treated group (O) showed a marked decrease in cytoplasmic and nuclear expression of the NFκB-P65 antibody (arrowhead). J The nano combined-treated group (Nano-O) showed a significant decrease in NFκB-P65 immunostaining within the hepatic cells (arrowheads). CCL₄ carbon tetrachloride, CS chitosan, GSH glutathione, Fe₃O₄ ferric oxide, VCO virgin coconut oil