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. 2016 Aug 6;7(3):416-27.
doi: 10.4292/wjgpt.v7.i3.416.

Effects of aging on the architecture of the ileocecal junction in rats

Maria Cícera de Brito  1 , Renato Paulo Chopard  1 , Diego Pulzatto Cury  1 , Ii Sei Watanabe  1 , Cristina Eusébio Mendes  1 , Patricia Castelucci  1
Affiliations

Effects of aging on the architecture of the ileocecal junction in rats

Maria Cícera de Brito et al. World J Gastrointest Pharmacol Ther. .

Abstract

Aim: To evaluate the structural organization of the elastic and collagen fibers in the region of the ileocecal transition in 30 young and old male Wistar rats.

Methods: Histology, immunohistochemistry (IHC), transmission electron microscopy and scanning electron microscopy were employed in this study. The results demonstrated that there was a demarcation of the ileocecal region between the ileum and the cecum in both groups.

Results: The connective tissue fibers had different distribution patterns in the two groups. IHC revealed the presence of nitric oxide synthase, enteric neurons and smooth muscle fibers in the ileocecal junctions (ICJs) of both groups. Compared to the young group, the elderly group exhibited an increase in collagen type I fibers, a decrease in collagen type III fibers, a decreased linear density of oxytalan elastic fibers, and a greater linear density of elaunin and mature elastic fibers.

Conclusion: The results revealed changes in the patterns of distribution of collagen and elastic fibers that may lead to a possible decrease in ICJ functionality.

Keywords: Aging; Collagen fibers; Elastic fibers; Ileocecal junction; Rats.

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Figures

Figure 1
Figure 1
Histological sections of the ileocecal junctions in the young group and the elderly group. Hematoxylin-eosin staining (A-D) was performed to examine the ileum (IL), cecum (C), circular muscle layer (CM) and longitudinal muscle layer (ML). Picrosirius staining under non-polarized light (E and G) showed the transition region between the ileum (IL) and cecum (C). Picrosirius staining under polarized light (F and H) showed type I collagen fibers (yellow, orange and red) (arrows) and type III collagen fibers (green) (arrowhead).
Figure 2
Figure 2
Histological sections of ileocecal junctions in the young group (A, C, E) and the elderly group (B, D, F) stained with Weigert with oxone (A, B), Weigert (C, D) and Verhoeff (E, F).
Figure 3
Figure 3
Immunohistochemistry mistry final. HuC/D staining in immunoreactive neurons (A, D), NOS staining in immunoreactive neurons and nerve fibers (B, E), and DAPI staining of the nuclei of immunoreactive cells (C, F) of the ileocecal junction in young rats (A-C) and elderly rats. The co-localization of HuC/D, NOS and DAPI (C, F). The arrows indicate immunoreactive neurons stained for HuC/D (A, D), NOS (B, E), and triple colocalization with HuC/D, NOS and DAPI (C, F). The double arrows indicate immunoreactive fibers stained for NOS (B, E). The double arrows show muscle fiber nuclei stained with DAPI (C, F). NOS: Nitric oxide synthage; DAPI: 4',6-diamidino-2-phenylindole.
Figure 4
Figure 4
Transmission electron microscopy. A: A cross-section of the muscle layer showing the muscle fibers (*) and connective tissue (arrow) of the young group. Magnification: × 6000; B: A longitudinal section of the muscle layer showing muscle fibers (*), nuclei (N) and a set of unmyelinated fibers (arrows) of the young group Magnification: × 20000; C: A longitudinal section of the muscular layer showing collagen fibers arranged in different directions (arrows) and muscle fibers of the elderly group. Magnification: × 10000; D: Nucleus (N) and collagen fibers arranged in different directions (arrows) and synaptic vesicles (arrowhead) of the elderly group. Magnification: × 10000.
Figure 5
Figure 5
Normal mucosa of the rat ileocecal transition point of the young and elderly groups. A: Normal mucosa of the rat ileocecal transition point of the young showing the ileum (IL) and cecum (*); B: Villi with a normal appearance (arrows); C: Sample treated with NaOH solution. Image showing the connective tissue with different forms of blades (*) and numerous foramens (arrows) in a network of collagen fibers; D: Image showing the interphase cell surface tissue of the cecum, including the foramen; E: Ileum (IL), cecum (*) and ileocecal junction (arrows) of the elderly group; F: The highest increase in ileal villi (arrows); G: Sample treated with NaOH solution. Ileum tissue-shaped blade (*) and numerous foramens as a network of collagen fibrils (arrows). Image showing the foramen cecum surface (arrows).
Figure 6
Figure 6
Quantitative analysis. Data related to the means ± SD of the mean linear (pixels) of type I and type III collagen (A) and data indicating the linear density (LD) expressed as the mean ± SD of oxytalan elastic fibers (Weigert) (μm2), elaunin fibers (Verhoeff) (μm2), and mature elastic fibers (Weigert-Oxane) (μm2) (B) of the young and old groups. bP < 0.001.
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