Anat Cell Biol.  2012 Dec;45(4):229-240. 10.5115/acb.2012.45.4.229.

Preliminary morphological and biochemical changes in rat liver following postnatal exposure to sodium arsenite

• Bhattacharya A
• Dhar P
• Mehra RD

• KMID: 2133909
• DOI: http://doi.org/10.5115/acb.2012.45.4.229

Abstract

The effects of sodium arsenite exposure on the hepatic maturation period of cellular and functional reorganization in developing rat livers were evaluated. Animals received intraperitoneal injections of sodium arsenite (1.5 mg/kg body weight) or distilled water on days 9 to 28 after birth. On day 29, the animals were sacrificed either by cervical dislocation or by perfusion fixation. The perfusion fixed liver tissue was processed for paraffin embedding, sectioning and hematoxylin and eosin staining. The fresh liver tissue was processed for cryo-sectioning followed by Sudan Black B staining and for biochemical estimation of reduced glutathione. Microscopic observation revealed comparable preserved hepatic lobular patterns and distributions of uninucleate and binucleate hepatocytes in the control and the experimental groups. The mean nuclear area and diameter of the hepatocytes was increased in the experimental group. Lipid droplet distribution pattern in Sudan Black B stained sections revealed higher staining intensity towards the centrilobular area in both groups. Semiquantitative estimation of staining intensity showed lower mean gray values in zone 3 than in zones 2 and 1 (suggestive of the setting in of the adult pattern) in both groups. The reduced glutathione levels in the liver tissue and the altered nuclear size of the hepatocytes in the experimental group suggested the impairment of morphological and biochemical processes induced by arsenic exposure during the postnatal period.

Keyword

Arsenic; Wistar rats; Liver; Morphometry; Sudan Black B; Reduced glutathione

MeSH Terms

Adult
Animals
Arsenic
Arsenites
Azo Compounds
Biochemical Processes
Dislocations
Eosine Yellowish-(YS)
Glutathione
Hematoxylin
Hepatocytes
Humans
Injections, Intraperitoneal
Liver
Paraffin Embedding
Parturition
Perfusion
Rats
Rats, Wistar
Sodium
Sodium Compounds
Sudan
Water
Arsenic
Arsenites
Azo Compounds
Eosine Yellowish-(YS)
Glutathione
Hematoxylin
Sodium
Sodium Compounds
Water

Figure

• 

  Fig. 1 Experimental design. GSH, glutathione; i.p., intra-peritoneal; PND, post natal days.

• 

  Fig. 2 High magnification photo micrographs of (A) hematoxylin and eosin stained and (B) Sudan Black B stained sections of liver (postnatal days [PND] 29). (A) A standard rectangular grid (270 µm × 80 µm) between the central vein (black asterisk) and the portal triad (black triangle). The nuclear area and equivalent diameter of the nuclei were generated by encircling the nuclear outline after selecting tool AREA (circle). Uninucleate (), binucleate () cells (NIS Elements AR 3.1 software). (B) Three rectangular grids of equal area between the central vein (red asterisk) and the inter portal triad region (red triangle) dividing the parenchyma centrifugally into zones (3, 2, 1), from which the mean gray value of each grid area and mean gray value per zone were measured. Scale bars=20 µm (A), 50 µm (B).

• 

  Fig. 3 Linear growth curve based on daily recorded weight (postnatal days [PND] 1 to 29) (A); bar diagram showing body weight at PND 9 (beginning of the treatment period) and PND 29 (day of sacrifice) (B) of the control and the experimental groups. Values expressed as mean±SD.

• 

  Fig. 4 Low (A, B) and high (C-F) magnification photomicrographs of hematoxylin and eosin stained sections of liver (postnatal days 29) showing: preserved classical lobular pattern (dotted outline) with the central vein (black asterisk) in the center and the portal triads (black triangles) at the corners of the lobule in (A) control and (B) experimental animals. Apparent increase in hepatocyte size () and dilatation of sinusoids () in experimental group (D). Single cell thick hepatocyte plates (circle), binucleate cell (), kupffer cell (), endothelial cell lining the central vein () (C, D), polyhedral hepatocytes (), centrally placed spherical nucleus (). Scale bars=200 µm (A, B), 50 µm (C, D), 20 µm (E, F).

• 

  Fig. 5 High magnification photo micrographs of hematoxylin and eosin stained sections of liver (postnatal days 29) from control (A) and experimental (B) group. Intracytoplasmic vacuolization () around the central vein (asterisk) in (B); sinusoids (). Sclae bars=50 µm (A, B).

• 

  Fig. 6 Bar diagrams of (A) mean nuclear area (µm2), (B) mean nuclear diameter (µm), and (C) percentage of uni and binucleate hepatocytes in different lobes of the liver of hepatocytes of the control and the experimental groups. Values expressed as mean±SD, *P<0.05.

• 

  Fig. 7 High magnification photo micrographs of Sudan black B stained sections of liver (postnatal days [PND] 29) from (A) the control and (B) the experimental groups. More () and less () intensely stained hepatocytes, intense staining of cytoplasm against pale nuclei (triangle), central vein (asterisks); sinusoids (). Scale bars=50 µm (A, B). (C) Bar diagram showing the mean grey values in the zones of hepatic acinus of the control and the experimental groups.

• 

  Fig. 8 Bar diagram showing the levels of reduced glutathione (GSH, µg/g of liver tissue) in the control and the experimental groups. Value expressed as mean±SD, *P<0.05.

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