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Long live the liver: immunohistochemical and stereological study of hepatocytes, liver sinusoidal endothelial cells, Kupffer cells and hepatic stellate cells of male and female rats throughout ageing

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Abstract

Male/female differences in enzyme activity and gene expression in the liver are known to be attenuated with ageing. Nevertheless, the effect of ageing on liver structure and quantitative cell morphology remains unknown. Male and female Wistar rats aged 2, 6, 12 and 18 months were examined by means of stereological techniques and immunohistochemical tagging of hepatocytes (HEP), liver sinusoidal endothelial cells (LSEC), Kupffer cells (KC) and hepatic stellate cells (HSC) in order to assess the total number and number per gram of these cells throughout life. The mean cell volume of HEP and HSC, the lobular position and the collagen content of the liver were also evaluated with stereological techniques. The number per gram of HSC was similar for both genders and was maintained throughout ageing. The mean volume of HSC was also conserved but differences in the cell body and lobular location were observed. Statistically significant gender differences in HEP were noted in young rats (females had smaller and more binucleated HEP) but were attenuated with ageing. The same occurred for KC and LSEC, since the higher number per gram in young females disappeared in older animals. Liver collagen increased with ageing but only in males. Thus, the numbers of these four cell types are related throughout ageing, with well-defined cell ratios. The shape and lobular position of HSC change with ageing in both males and females. Gender dimorphism in HEP, KC and LSEC of young rat liver disappears with ageing.

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Authors and Affiliations

  1. Laboratory of Histology and Embryology, Department of Microscopy, Institute of Biomedical Sciences Abel Salazar, University of Porto, ICBAS-UP, Rua de Jorge Viterbo Ferreira no. 228, 4050-313, Porto, Portugal

    Ricardo Marcos

  2. Scotland’s Rural College (SRUC), Epidemiology Research Unit—Future Farming Systems Group, Inverness, UK

    Carla Correia-Gomes

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  1. Ricardo Marcos
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  2. Carla Correia-Gomes
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Correspondence to Ricardo Marcos.

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Supplemental Fig. 1

Thick section of an adult male liver immunostained against glial fibrillary acidic protein. Hepatic stellate cells (arrows) can be seen around the portal tract (P) and central vein (V). For unequivocal identification of the location, glutamine synthetase immunomarking (tags pericentral hepatocytes) was also used. Bar 30 μm. (GIF 3592 kb)

High resolution image (TIFF 9271 kb)

Supplemental Figure 2

Thin sections of old male (a) and old female (b) rat liver immunostained against α-smooth muscle actin. Immunomarking is restricted to the wall of blood vessels in portal tract (a) and in the central vein (b), with no noticeable positive cells alongside the sinusoids. Bar 30 μm. (GIF 2692 kb)

High resolution image (TIFF 7509 kb)

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Marcos, R., Correia-Gomes, C. Long live the liver: immunohistochemical and stereological study of hepatocytes, liver sinusoidal endothelial cells, Kupffer cells and hepatic stellate cells of male and female rats throughout ageing. Cell Tissue Res 366, 639–649 (2016). https://doi.org/10.1007/s00441-016-2490-y

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