Hepatic and immune modulatory effectiveness of lactoferrin loaded Selenium nanoparticles on bleomycin induced hepatic injury

Abstract

This study aimed to estimate the hepatic and immune ameliorating potential of extracted bovine lactoferrin (LF), Selenium nanoparticles (SeNPs) or their combination (LF/SeNPs) against bleomycin (BLM) induced hepatic injury. Fifty adult male rats (160-200 g) were equally divided into five groups: (1) the saline control group, (2) BLM-injected (15 mg/kg twice a week, ip), and (3-5) groups treated orally with LF (200 mg/kg/day), SeNPs (0.0486 mg/kg/day) or LF/SeNPs combination (200.0486 mg/kg/day) for 6 weeks post BLM-intoxication. Blood and liver samples were subjected to biochemical, histopathological, and immunohistochemical analyses. The results revealed that BLM caused a significant increase in hepatic lipid peroxidation and nitric oxide, as well as serum markers of liver functions (AST, ALT and GGT activities), and levels of GM-CSF, CD4, TNF-α, IL-1β, TGF-β1, fibronectin, triglycerides, cholesterol and LDL-C. Additionally, hepatic glutathione, Na⁺/K⁺-ATPase, and glutathione peroxidase, as well as serum HDL-C, total protein and albumin levels were significantly reduced. Moreover, BLM injection resulted in marked histopathological alterations and severe expression of caspase 3. Post-treatment of BLM-intoxicated rats with LF, SeNPs or LF/SeNPs combination obviously improved the BLM-induced hepatic damages; this was achieved from the marked modulations in the mentioned parameters, besides improving the histopathological hepatic architecture. It is worth mentioning that LF/SeNPs exerted the greatest potency. In conclusion, the obtained results demonstrated that LF, SeNPs and LF/SeNPs succeeded in attenuating the BLM-induced hepatic dysfunction. Therefore, these supplements might be used to protect against drug-associated side effects.

Keywords: Bleomycin; Fibrosis; Lactoferrin; Liver; Nano-Selenium; Rats.

Fig. 1

Chromatogram of the purified lactoferrin extracted from raw bovine milk.

Fig. 2

Effect of different treatments on serum CD4, GM-CSF, TNF-α, IL-1β, TGF-β1, and fibronectin levels of BLM-intoxicated rats. * Is significant versus the control group, while # is significant versus BLM group at p ≤ 0.05. LF (lactoferrin); BLM (bleomycin); SeNPs (Selenium nanoparticles).

Fig. 3

Photomicrographs of liver tissues of the different study groups. The anti-caspase-3 immunoreactivity represented in the tissues as brown color by DAB chromogen. (A) The control group showed weak cytoplasmic caspase-3 immunoreactivity; (B) The BLM-injected group showed severe caspase-3 immunoreactivity; (C) The BLM ~ LF group showed mild caspase-3 immunoreactivity; (D) The BLM ~ SeNPs group showed moderate caspase-3 immunoreactivity; (E) The BLM ~ LF/SeNPs group showed a weak, approximately normal, caspase-3 positivity.

Fig. 4

Intensity quantitative measurement for positive caspase-3 immune reactions. Data are presented as mean ± standard error; subjected to one-way ANOVA followed by post hoc (Duncan) test at p ≤ 0.05; * is significant versus the control group, while # is significant versus BLM group; LF (lactoferrin); BLM (bleomycin); SeNPs (Selenium nanoparticles).

Fig. 5

Photomicrographs of liver tissues of the different study groups; (A) Control group showing normal hepatic lobule structure with central vein (*), hepatic cord (Hc), binucleated cell (short arrow) and hepatic sinusoids with epithelial cells (arrowhead); (B) BLM-intoxicated group showed focal necrosis with pycnotic nuclei (*), congested bile duct (long arrow) and obliterated sinusoids (short arrow); (C) BLM ~ LF treated group showed focal necrosis (*), and congested sinusoids (long arrow); (D) BLM ~ SeNPs treated group showed congested sinusoids (long arrow) with normal hepatic pattern; (E) BLM ~ LF/SeNPs formulation treated group showed approximately normal architecture.