Figure 1.
Study of morphological mucosal changes of the small bowel in NEC-induced in rats pups.
(a) Histological analysis in the three pups groups (group 1: full term and were mother-fed; group 2: preterm and mother-fed; group 3: preterm, hand-gavaged with a protein rich formula after a hypoxia–reoxygenation procedure). according to the histological classification as showing in (b): intact morphology of the villi (a); sloughing of villi tips (b); mild-villous necrosis (c); loss of villi (d); complete destruction of the mucosa, transmural necrosis (e). (c) Severity of NEC in the different pups groups. Original magnifications: 120×(a,b,c,d), 60×(e).
Figure 2.
Immunohistochemical features of epithelial cells positive for anti Ki-67 antibody staining in the small bowel mucosa.
(a) normal staining is limited to the crypts; (b) abnormal staining extends beyond the crypts, and is continuous, irregular or spreads into the covering villi (arrowheads). (c) Histological analysis of KI-67 localisation. Representative images are shown (original magnification 400×)
Figure 3.
Scales used for the semiquantitation of the small bowel mucosal epithelial cells positive for immunohistochemical staining with an anti caspase-3 antibody.
(0): with 0 to 2 cells; (1): with 2 to 5 cells; (2): with 5 to 20 cells; (3): with 20 to 50 cells; (4): more than 50 cells. Representative images are shown (original magnification 400×).
Figure 4.
Immunohistochemistry using anti-TLR 2 antibody in ileal samples.
(a) In control group, TLR-2 positive cells were located in the crypts, with weak and apical cytoplasmic staining. (b) In group 2 (scored as scale (b) in H&E-stained sections) and (c) in group 3 combining the 3 factors (scored as scale (d) in H&E-stained sections), a strong cytoplasmic TLR-2 staining was observed in the crypts and in the villi (Original magnification 200×, 400×).
Figure 5.
Immunohistochemistry using anti anti-pNF-κB antibody in ileal samples.
In group 1, there is no nuclear staining in an intestinal villous (a and e) whereas in group 2 (scored as scale (b) in H&E-stained sections), some positive cells are observed (b). These positive cells are much more numerous in the group 3 (scored as scale (c) in H&E-stained section) (c and d). (Original magnification ×400 and 800 respectively).
Figure 6.
mRNA expression levels of TLR-2 and TLR-4 from distal jejunum were analysed by real-Time-PCR under basal and NEC conditions.
(a) TLR-2 and (b) TLR-4 mRNA expression levels from distal jejunum. Data represent the means±SEM of 10 mice per group. *P<0.05 and **P<0.01, significantly different from group 1.