Anat Cell Biol.  2023 Dec;56(4):538-551. 10.5115/acb.23.101.

A comparative study on the hepatoprotective effect of selenium-nanoparticles and dates flesh extract on carbon tetrachloride induced liver damage in albino rats

• Ouais GN
• Hassan DM

• KMID: 2548915
• DOI: http://doi.org/10.5115/acb.23.101

Abstract

Exposure to environmental pollutants such as carbon tetrachloride (CCL 4 ) causes liver damage. This study aimed to compare the ameliorative activity of the dates flesh extract (DFE) and selenium-nanoparticles (SeNPs) on CCL 4 -induced hepatotoxicity and if DFE could be a useful alternative supplement. Twenty-four male albino rats were enrolled and randomly divided into four equal groups (6 rats in each group): control group received only basal diet with no medications. Group II received CCL 4 in a dose of 0.5 mg/kg intraperitoneal injection twice weekly for four weeks. Group III rats were pretreated with SeNPs in a dose of 2.5 mg/kg once a day orally three times/wk for four weeks alone then combined with the previously described dose of CCL 4 for another four weeks. Group IV rats were pretreated with DFE in a dose of 8 ml of the aqueous extract/kg/d orally for four weeks alone then combined with the previously described dose of CCL 4 for another four weeks. The liver damage was assessed by estimation of plasma concentration of albumin and enzymes activities of alanine aminotransferase and tissue genes expression. Liver oxidation levels were assessed by measuring the tissue concentration of the malondialdehyde, superoxide dismutase, and the total glutathione. Additionally, inflammatory mediators tumour necrosis factor--α and interleukin-6 were estimated. Detecting the liver’s cellular structural damage was done by histopathological and immunohistochemical examination. This study suggests that CCL 4 -induced liver damage in rats can be protected by administration whether the costly SeNPs or the economical DFE.

Keyword

Selenium-nanoparticles; Dates-flesh-extract; Hepatoprotection; Carbon tetrachloride; Vitamin E

Figure

• 

  Fig. 1 The mean levels of liver functions: (A) ALT [U/L) and (B) albumin [g/dl) were assessed in all studied groups. *Significant difference versus control group, #significant difference versus CCL4 group (P-value<0.05). ALT, alanine aminotransferase; CCL4, carbon tetrachloride; SeNPs, selenium-nanoparticles; DFE, dates flesh extract.

• 

  Fig. 2 Enzymatic activity of (A) SOD (U/ml), (B) GSH-px (mU/ml), and (C) MDA (nmol/ml) in liver tissue of all studied groups. *Significant difference versus control group, #significant difference versus CCL4 group (P-value<0.05). SOD, superoxide dismutase; GSH- px, glutathione peroxidase; MDA, malondialdehyde; CCL4, carbon tetrachloride; SeNPs, selenium-nanoparticles; DFE, dates flesh extract.

• 

  Fig. 3 Mean of serum levels of inflammatory markers (A) TNF-α (pg/mg) and (B) IL-6 (pg/mg). *Significant difference versus control group, #significant difference versus CCL4 group (P-value<0.05). TNF-α, tumor necrosis factor-α; IL-6, interleukin-6; CCL4, carbon tetrachloride; SeNPs, selenium-nanoparticles; DFE, dates flesh extract.

• 

  Fig. 4 Mean of genes expression of (A) TGF-β1, (B) HGF, and (C) MMP-2 in liver tissue of all studied groups. *Significant difference versus control group, #significant difference versus CCL4 group (P-value<0.05). TGF-β1, transforming growth factor-β1; HGF, hepatocellular growth factor; MMP-2, matrix metalloproteinase; CCL4, carbon tetrachloride; SeNPs, selenium-nanoparticles; DFE, dates flesh extract.

• 

  Fig. 5 H&E-stained liver sections; (A, B) Control group, the hepatic central vein (CV) with polygonal hepatocytes (H) shows acidophilic cytoplasm and vesicular nucleus. Thin wall blood sinusoids (S) radiating between hepatocytes cords with their lining endothelium was seen. (C) CCL4 group, the hepatocytes show disorganization of hepatic cords. The portal regions show cellular degeneration (arrows), some hepatocytes showed pyknotic nuclei (arrow heads) while others have vesicular nuclei (H). Some hepatocytes show rarefication of their cytoplasm (curved arrows) while others have cytoplasmic vacuolations (V) and hemorrhage (stars). (D) Another section in the same group shows marked dilated congested vein in the periportal and pericentral areas (star). Also, cellular degeneration with inflammatory infiltrate (IF) in the portal region (arrows) was seen. (E, F) SeNPs group, the hepatocytes show more or less normal architecture (arrowheads) with normal hepatic cords. Note small dark pyknotic nuclei (arrows) and mild congestion of the CV. (G, H) DFE group, effectively cessation of inflammatory response with the improvement of hepatocytic degeneration and regain of the normal architecture excluding some areas (arrows). Some hepatocytes show cytoplasmic vacuolations (V). CCL4, carbon tetrachloride; SeNPs, selenium-nanoparticles.

• 

  Fig. 6 Masson’s trichrome-stained liver sections; (A) Control group, show collagen fibers normally distributed (arrows) around the portal tract. (B, C) CCL4 group, shows a marked increase in collagen fibers (arrows) deposition around the blood vessels in the portal area. (D, E) SeNPs group, shows minimal collagen fibers deposition (long arrows) around the blood vessels in the portal area and between cells (short arrows). (F, G) DFE group, shows decreased collagen fibers deposition (arrows) around the blood vessels in the portal area. (H) Statistical analysis of the area percentage of collagen fibers deposition shows *significant difference against the control group. #Represents a significant difference against the CCL4 group. CCL4, carbon tetrachloride; SeNPs, selenium-nanoparticles; DFE, dates flesh extract.

• 

  Fig. 7 Immunohistochemical staining with α-SMA of (A, B) control group, most of the hepatocytes show negative reaction except few with mild cytoplasmic immune reaction. (C, D) CCL4 group, most hepatocytes have an intense immune reaction in hepatocytes and in-between hepatic lobules. (E, F) SeNPs group, shows a minimal immune reaction in vascular walls (arrows) and very mild reaction in between hepatic lobules. (G, H) DFE group, show mild to a moderate immune reaction in between hepatic lobules and among the hepatocytes (arrows). (I) Statistical analysis of the percentage of α-SMA positive reactions shows *significant difference against the control group. #Represents a significant difference against the CCL4 group. α-SMA, α-smooth muscle actin; CCL4, carbon tetrachloride; SeNPs, selenium-nanoparticles; DFE, dates flesh extract.

Reference

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