Gastroenterology Research, ISSN 1918-2805 print, 1918-2813 online, Open Access
Article copyright, the authors; Journal compilation copyright, Gastroenterol Res and Elmer Press Inc
Journal website http://www.gastrores.org

Original Article

Volume 6, Number 4, August 2013, pages 124-133


Oral Administration of Interleukin-10 and Anti-IL-1 Antibody Ameliorates Experimental Intestinal Inflammation

Figures

Figure 1.
Figure 1. Protective effects of interleukin-10/anti-IL-1 mix in chemical-induced chronic colitis. (A) Evaluation of colon resistance (expressed as Ω * cm2) by Ussing chamber analysis in different treatment groups. UNTR (untreated, hydroalcoholic solution only); DSS (DSS 2% plus hydroalcoholic solution); DSS + IL-10/anti-IL-1 (DSS 2% + interleukin-10 IL-10 plus anti-IL-1 antibody). The graph represents mean values ± S.D. (n = 10) Statistical analysis: DSS + IL-10/anti-IL-1 vs. DSS ** P = 0.0028. (B) Evaluation of the histologic score in different treatment groups. UNTR (untreated, hydroalcoholic solution only); DSS (DSS 2% plus hydroalcoholic solution); DSS + IL-10/anti-IL-1 (DSS 2% + interleukin-10 plus anti-IL-1 antibody). The graph represents mean values ± S.D. (n = 10) Statistical analysis: DSS + IL-10/anti-IL-1 vs. DSS ** P = 0.0062.
Figure 2.
Figure 2. Ematoxilin-Eosin staining of colon sections in chemical-induced chronic colitis. A: histological analysis of paraffin embedded colon sections. UNTR (untreated, hydroalcoholic solution only); B: DSS (DSS 2% plus hydroalcoholic solution); C: DSS + IL-10/anti-IL-1 (DSS 2% + interleukin-10 plus anti-IL-1 antibody).
Figure 3.
Figure 3. Evaluation of cytokine secretion in chemical-induced chronic colitis. ELISA assays for concentrations of TNF-alpha (A), IFN-gamma (B), IL-12 (C), KC (D), IL-17 (E) and IL-10 (F) in different treatment groups. UNTR (untreated, hydroalcoholic solution only); DSS (DSS 2% plus hydroalcoholic solution); DSS + IL-10/anti-IL-1 (DSS 2% + interleukin-10 plus anti-IL-1 antibody); data are represented as mean values ± S.D. (n = 10). Statistical analysis: A) DSS+ IL-10/anti-IL-1 vs. DSS ** P = 0.0036; B) DSS + IL-10/anti-IL-1 vs. DSS *** P = 0.004; C) DSS+ IL-10/anti-IL-1 vs. DSS ** P = 0.0053; D) DSS + IL-10/anti-IL-1 vs. DSS *** P = 0.0001; E) DSS + IL-10/anti-IL-1 vs. DSS * P = 0.036; F) DSS + IL-10/anti-IL-1 vs. DSS *** P = 0.0001.
Figure 4.
Figure 4. Evaluation of ZO-1 expression. Immunofluorescence anti-ZO-1 (red staining) experiments in the colon samples of different treatment groups showed the different epithelial barrier TJ structural integrity (arrows). A relevant example for each treatment group is shown: A (UNTR; untreated, hydroalcoholic solution only); B (DSS; DSS 2% plus hydroalcoholic solution); C (DSS + IL-10/anti-IL-1; DSS 2% + interleukin-10 plus anti-IL-1 antibody) Bars: 20 µm.
Figure 5.
Figure 5. Evaluation of occludin expression. Immunofluorescence anti-occludin (red staining) experiments in the colon samples of different treatment groups showed the different epithelial barrier TJ structural integrity. A relevant example for each treatment group is shown: A (UNTR; untreated, hydroalcoholic solution only); B (DSS; DSS 2% plus hydroalcoholic solution); C (DSS + IL-10/anti-IL-1; DSS 2% + interleukin-10 plus anti-IL-1 antibody). Bars: 20 µm.

Tables

Table 1. Treatment Scheme for Induction of the Chronic Colitis Model, in the Different Treatment Groups
 
Groups CyclesGroup 1: UNTR (10 mice)Group 2: DSS (10 mice)Group 3: DSS + IL-10/anti-IL-1 (10 mice)
Cycle 1 dd 1 - 7Control group: only normal drinking waterDSS 2% in waterDSS 2% in water
Interval dd 8 - 21Normal drinking waterNormal drinking water
Cycle 2 dd 22 - 28DSS 2% in waterDSS 2% in water
Interval dd 29 - 42Normal drinking waterNormal drinking water
Cycle 3 dd 43 - 49DSS 2% in waterDSS 2% in water
Interval dd 50 - 58Normal drinking waterNormal drinking water
Treatment dd 59 - 68 (Twice a day)Hydro-alcoholic solution vehicleHydro-alcoholic solution vehicle50 fg/kg GUNA interleukin-10 plus 50 fg/kg GUNA anti-IL-1 per os

 

Table 2. Scoring System for the Comparative Analysis of Intestinal Bleeding
 
Intestinal bleeding scoreStool consistencyBlood presence
0NormalNegative
1Soft but still formedPositive hemoccult
2Very softBlood traces in stool visible
3DiarrhoeaRectal bleeding

 

Table 3. Scoring System for Inflammation-Associated Histological Changes in the Colon
 
Histological scoreHistologic changes
0No evidence of inflammation
1Low level of inflammation with scattered infiltrating mononuclear cells (1-2 foci)
2Moderate inflammation with multiple foci
3High level of inflammation with increased vascular density and marked wall thickening
4Maximal severity of inflammation with transmural leukocyte infiltration and loss of goblet cells