Copyright
©The Author(s) 2019.
World J Gastroenterol. Jul 21, 2019; 25(27): 3572-3589
Published online Jul 21, 2019. doi: 10.3748/wjg.v25.i27.3572
Published online Jul 21, 2019. doi: 10.3748/wjg.v25.i27.3572
Table 1 Composition of the experimental diet
Ingredient, g/kg | P14 |
Acid casein (Armor Protéines®, ref. 139860) | 112 |
Whey protein (Armor Protéines®, Protarmor 80, ref. 139805) | 28 |
Corn starch | 622.4 |
Sucrose | 100.3 |
Cellulose | 50 |
Soybean oil | 40 |
Mineral mixture (AIN 93-M) | 35 |
Vitamin mixture (AIN 93-V) | 10 |
Choline | 2.3 |
Metabolizable energy, kJ/g | 14.5 |
Table 2 Colon morphometric and inflammatory markers
Parameter | d0 | d7 | d7 vs d0 | d10 | d10 vs d7 | d13 | d13 vs d10 | d28 | d28 vs d0 | Statistical effect (time) |
Colon length (cm) | 6.5 ± 0.2 | 4.9 ± 0.3a | ↓ | 5.4 ± 0.2a | - | 5.5 ± 0.2a | - | 5.7 ± 0.2a | ↓ | < 0.001 |
Colon weight/length (mg/cm) | 16 ± 0.3 | 24 ± 0.4a | ↑ | 31 ± 3.0ab | ↑ | 34 ± 3.0ab | - | 38 ± 8.0ab | ↑ | < 0.05 |
Osmolarity of colonic content | 180 ± 8 | 311 ± 15a | ↑ | 278 ± 8.0a | - | 288 ± 13a | - | 281 ± 15a | ↑ | < 0.01 |
Colonic water content (%) | 64.6 ± 2.5 | 87.5 ± 2.0a | ↑ | 75.5 ± 3.0ab | ↓ | 76.3 ± 1.5ab | - | 74.0 ± 1.9ab | ↑ | < 0.001 |
IL-1β (ng/mg of total colon protein) | 9.4 ± 1.8 | 14.3 ± 2.4a | ↑ | 13.7 ± 1.8a | - | 16.2 ± 2.5a | - | 15.4 ± 2.2a | ↑ | < 0.01 |
IL-6 (ng/mg of total colon protein) | 1.5 ± 0.5 | 3.6 ± 0.6a | ↑ | 4.2 ± 0.8a | - | 4.9 ± 1.2a | - | 2.3 ± 0.3bcd | - | < 0.001 |
MPO activity (UA/mg total colon protein) | 1.2 ± 0.2 | 5.8 ± 1.2a | ↑ | 5.3 ± 0.8a | - | 3.1 ± 0.8abc | ↓ | 1.9 ± 0.6bcd | - | < 0.001 |
Plasmatic LBP (µg/mL) | 2.0 ± 0.2 | 5.8 ± 0.4a | ↑ | 4.5 ± 0.5ab | ↓ | 4.9 ± 0.6ab | - | 4.6 ± 0.3c | ↑ | < 0.001 |
Table 3 Histological analysis scoring of inflammatory and healing parameters
Parameter | d7 | d10 | d10 vs d7 | d13 | d13 vs d10 | d28 | d28 vs d7 | Statistical effect (time) |
Inflammatory score | 12.8 ± 0.8 | 9.3 ± 1.4b | ↓ | 8.4 ± 1.3b | - | 4.4 ± 1.7bcd | ↓ | < 0.001 |
Goblet cell depletion | 2.27 ± 0.24 | 1.83 ± 0.21 | - | 1.08 ± 0.26bc | ↓ | 0.88 ± 0.30bc | ↓ | < 0.001 |
Ulceration and erosion | 2.09 ± 0.29 | 1.34 ± 0.31 | - | 1.17 ± 0.27 | - | 0.63 ± 0.32bcd | ↓ | < 0.001 |
Crypt damage | 2.72 ± 0.19 | 1.58 ± 0.34b | ↓ | 1.25 ± 0.30b | - | 1.25 ± 0.45b | ↓ | < 0.001 |
Edema | 2.09 ± 0.09 | 1.25 ± 0.22b | ↓ | 1.42 ± 0.23b | - | 0.0 ± 0.0bcd | ↓ | < 0.001 |
Inflammatory infiltrate (mononuclear cells) | 1.91 ± 0.27 | 1.83 ± 0.30 | - | 1.75 ± 0.18 | - | 1.40 ± 0.33bcd | - | < 0.001 |
Inflammatory infiltrate (neutrophils) | 1.73 ± 0.13 | 1.50 ± 0.29 | - | 1.75 ± 0.19 | - | 0.63 ± 0.32bcd | ↓ | < 0.001 |
Epithelial repair score | 0.82 ± 0.22 | 2.83 ± 0.42b | ↑ | 3.25 ± 0.43b | - | 3.25 ± 0.25b | ↑ | < 0.001 |
Gland hyperplasia and presence of mitotic cells | 0.0 ± 0.0 | 0.83 ± 0.24b | ↑ | 0.92 ± 0.26b | - | 0.88 ± 0.35b | ↑ | < 0.001 |
Re-epithelialization | 0.54 ± 0.16 | 0.58 ± 0.15 | - | 0.58 ± 0.15 | - | 0.63 ± 0.33 | - | NS |
Crypt repair | 0.27 ± 0.19 | 1.42 ± 0.34b | ↑ | 1.75 ± 0.30b | - | 1.75 ± 0.45b | ↑ | < 0.001 |
Table 4 Kinetics of colonic mRNA expression level normalized to day 7 group (2-ΔΔCt)
d0 | d7 | d7 vs d0 | d10 | d10 vs d7 | d13 | d13 vsd10 | d28 | d28 vsd0 | Statistical effect (time) | |
Tight-junction protein | ||||||||||
Cldn2 | 2.41 ± 0.29 | 1.24 ± 0.21a | ↓ | 2.90 ± 0.15b | ↑ | 1.94 ± 0.17b | - | 2.26 ± 0.31b | - | < 0.001 |
Tjp1 | 1.45 ± 0.09 | 1.03 ± 0.08a | ↓ | 1.17 ± 0.10b | ↑ | 1.30 ± 0.06b | - | 1.29 ± 0.06b | - | < 0.01 |
Ocln | 2.92 ± 0.37 | 1.18 ± 0.24a | ↓ | 1.36 ± 0.11a | - | 1.96 ± 0.18abc | ↑ | 2.21 ± 0.30bc | - | < 0.001 |
ECM remodeling | ||||||||||
Mmp7 | 0.58 ± 0.16 | 1.46 ± 0.40a | ↑ | 5.31 ± 1.37ab | ↑ | 5.21 ± 1.22ab | - | 3.69 ± 0.85ab | - | < 0.001 |
Mmp9 | 0.13 ± 0.01 | 1.35 ± 0.24a | ↑ | 1.17 ± 0.27a | - | 0.89 ± 0.15a | - | 0.55 ± 0.20bcd | - | < 0.001 |
Timp1 | 0.05 ± 0.00 | 1.75 ± 0.49a | ↑ | 0.84 ± 0.25ab | ↓ | 0.41 ± 0.05b | - | 0.23 ± 0.06bc | - | < 0.001 |
Col3a1 | 0.77 ± 0.07 | 1.60 ± 0.33a | ↑ | 1.15 ± 0.15a | - | 1.20 ± 0.12a | - | 0.83 ± 0.16bcd | - | < 0.01 |
Wisp | 0.31 ± 0.02 | 1.40 ± 0.35a | ↑ | 0.78 ± 0.17ab | ↓ | 0.52 ± 0.04b | - | 0.43 ± 0.06b | - | < 0.001 |
Acta2 | 1.33 ± 0.17 | 1.25 ± 0.22 | - | 0.95 ± 0.16 | - | 0.79 ± 0.05a | - | 0.85 ± 0.11 | - | < 0.01 |
Goblet cell markers and mucins | ||||||||||
Tff3 | 1.80 ± 0.24 | 1.29 ± 0.32 | - | 1.05 ± 0.12ad | - | 1.62 ± 0.19c | ↑ | 1.87 ± 0.13c | - | < 0.05 |
Muc2 | 3.77 ± 0.64 | 1.10 ± 0.16a | ↓ | 1.72 ± 0.22a | - | 3.03 ± 0.18bc | ↑ | 3.38 ± 0.21bc | - | < 0.001 |
Klf4 | 2.31 ± 0.13 | 1.12 ± 0.22a | ↓ | 1.52 ± 0.15a | - | 1.73 ± 0.10a | - | 2.56 ± 0.24bcd | - | < 0.001 |
Epithelial repair modulating factors | ||||||||||
Igf-1 | 0.27 ± 0.02 | 1.23 ± 0.19a | ↑ | 1.14 ± 0.22ad | - | 0.77 ± 0.06ac | - | 0.55 ± 0.09abcd | - | < 0.001 |
Il-15 | 4.59 ± 0.89 | 1.32 ± 0.51a | ↓ | 1.67 ± 0.18a | - | 3.61 ± 0.91bc | ↑ | 4.73 ± 0.86bc | - | < 0.01 |
Il-22 | 0.33 ± 0.20 | 1.59 ± 0.44a | ↑ | 0.36 ± 0.08b | ↓ | 0.47 ± 0.14b | - | 0.18 ± 0.04b | - | < 0.001 |
Il-33 | 0.08 ± 0.02 | 1.69 ± 0.43a | ↑ | 0.60 ± 0.11ab | ↓ | 0.43 ± 0.05ab | - | 0.26 ± 0.06ab | ↑ | < 0.001 |
Saa | 1.51 ± 0.27 | 1.21 ± 0.23 | - | 1.05 ± 0.16 | - | 3.40 ± 0.40abc | ↑ | 3.18 ± 0.31abc | ↑ | < 0.001 |
Tgf-ß1 | 0.50 ± 0.03 | 1.29 ± 0.28a | ↑ | 0.72 ± 0.07b | ↓ | 0.51 ± 0.07b | - | 0.43 ± 0.04b | - | < 0.001 |
Tgf-ß3 | 1.08 ± 0.11 | 1.47 ± 0.37 | - | 0.72 ± 0.10b | ↓ | 0.93 ± 0.12 | - | 1.16 ± 0.16 | - | < 0.01 |
Inflammatory markers | ||||||||||
Il-1ß | 0.04 ± 0.01 | 1.71 ± 0.50a | ↑ | 0.39 ± 0.06ab | ↓ | 0.45 ± 0.07ab | - | 0.39 ± 0.09ab | ↑ | < 0.001 |
Il-6 | 0.01 ± 0.00 | 1.73 ± 0.51a | ↑ | 0.84 ± 0.28ab | ↓ | 0.41 ± 0.08b | - | 0.15 ± 0.07b | - | < 0.001 |
Il-10 | 0.23 ± 0.03 | 1.37 ± 0.27a | ↑ | 0.37 ± 0.04b | ↓ | 0.31 ± 0.06b | - | 0.25 ± 0.03b | - | < 0.001 |
Il-13 | 3.81 ± 0.75 | 1.26 ± 0.23a | ↓ | 0.82 ± 0.08a | - | 0.78 ± 0.10a | - | 2.53 ± 0.39bcd | - | < 0.001 |
Tnf-α | 0.13 ± 0.01 | 1.29 ± 0.22a | ↑ | 0.46 ± 0.05ab | ↓ | 0.50 ± 0.06ab | - | 0.47 ± 0.07ab | ↑ | < 0.001 |
Ptgs | 0.14 ± 0.01a | 1.69 ± 0.39b | ↑ | 0.84 ± 0.30c | ↓ | 0.60 ± 0.10 c | - | 0.29 ± 0.05a | - | < 0.001 |
Table 5 Mucosa-adherent microbiota composition and cecal bacterial metabolic activity
Parameter | d0 | d7 | d7 vs d0 | d10 | d10 vs d7 | d13 | d13 vs d10 | d28 | d28 vs d0 | Statistical effect (time) | |
Total bacteria (log/g mucosal content) | 9.36 ± 0.35 | 10.4 ± 0.14a | ↑ | 9.92 ± 0.16b | ↓ | 10.6 ± 0.22ac | ↑ | 10.2 ± 0.24ac | ↑ | < 0.001 | |
Shannon index | 6.46 ± 0.07 | 4.75 ± 0.32a | ↓ | 4.56 ± 0.28a | - | 4.44 ± 0.26a | - | 5.03 ± 0.29a | ↓ | < 0.001 | |
Chao index | 794 ± 55.2 | 633 ± 43.7a | ↓ | 593 ± 31.1a | - | 635 ± 34.4a | - | 606 ± 51.6a | ↓ | < 0.01 | |
Actinobacteria (%) | |||||||||||
Bifidobacteriaceae | 0.02 ± 0.01 | 0.01 ± 0.00 | - | 0.01 ± 0.01 | - | 0.00 ± 0.00 | - | 0.60 ± 0.28abcd | ↑ | < 0.001 | |
Coriobacteriaceae | 0.01 ± 0.02 | 0.04 ± 0.02 | - | 0.07 ± 0.04 | - | 0.06 ± 0.04 | - | 0.83 ± 0.31abcd | ↑ | < 0.001 | |
Bacteroidetes (%) | |||||||||||
Bacteroidaceae | 4.35 ± 1.24 | 5.6 ± 1.34 | - | 11.55 ± 2.77b | ↑ | 3.08 ± 1.08c | ↓ | 20.1 ± 4.98cd | ↑ | < 0.05 | |
Porphyromonadaceae | 12.1 ± 2.55 | 7.40 ± 2.08 | - | 2.53 ± 1.29 | - | 1.03 ± 0.35ab | - | 12.1 ± 5.18d | - | < 0.001 | |
Prevotellaceae | 1.06 ± 0.33 | 0.01 ± 0.00a | ↓ | 0.01 ± 0.01a | - | 0.02 ± 0.01a | - | 0.01 ± 0.01a | ↓ | < 0.01 | |
Alloprevotella | 1.04 ± 0.32 | 0.00 ± 0.00a | ↓ | 0.01 ± 0.01a | - | 0.01 ± 0.01a | - | 0.00 ± 0.00a | ↓ | < 0.05 | |
Rikenelleceae | 9.64 ± 1.79 | 0.33 ± 0.24a | ↓ | 0.33 ± 0.24a | - | 0.31 ± 0.26a | - | 2.04 ± 1.20a | ↓ | < 0.001 | |
Alistipes | 9.48 ± 0.72 | 1.04 ± 0.51a | ↓ | 0.24 ± 0.15a | - | 0.13 ± 0.09a | - | 1.98 ± 1.20a | ↓ | < 0.001 | |
Deferribacterres (%) | ↓ | ||||||||||
Deferribacteraceae | Mucispirillum | 3.53 ± 1.86 | 9.98 ± 2.33a | ↑ | 10.16 ± 3.04a | - | 14.82 ± 2.80a | - | 3.49 ± 0.76bcd | - | < 0.001 |
Firmicutes (%) | |||||||||||
Clostridiaceae | Roseburia | 0.80 ± 0.17 | 0.01 ± 0.00a | ↓ | 0.01 ± 0.01a | - | 0.01 ± 0.01a | - | 0.03 ± 0.01a | ↓ | < 0.001 |
Clostridium XIVa | 6.07 ± 0.84 | 1.09 ± 0.35a | ↓ | 5.56 ± 2.73b | ↑ | 8.54 ± 2.80abc | ↑ | 4.01 ± 1.64b | - | < 0.05 | |
Butyrivibrio | 0.28 ± 0.21 | 0.00 ± 0.00a | ↓ | 0.00 ± 0.00a | - | 0.00 ± 0.00a | - | 0.00 ± 0.00a | ↓ | < 0.01 | |
Eubacteriaceae | 0.17 ± 0.05 | 0.04 ± 0.02a | ↓ | 0.00 ± 0.00a | - | 0.00 ± 0.00a | - | 0.00 ± 0.00a | ↓ | < 0.001 | |
Lachnospiraceae | 35.0 ± 4.85 | 13.5 ± 2.91a | ↓ | 21.7 ± 4.25a | - | 23.2 ± 4.49a | - | 23.6 ± 4.69 | - | < 0.01 | |
Acetitafactor | 5.46 ± 1.47 | 1.28 ± 0.55a | ↓ | 0.30 ± 0.13a | - | 1.11 ± 0.63a | - | 1.00 ± 0.63a | ↓ | < 0.001 | |
Lactobacillaceae | 0.42 ± 0.19 | 0.24 ± 0.12 | - | 3.41 ± 2.82 | - | 0.63 ± 0.26 | - | 1.44 ± 0.88 | - | NS | |
Ruminococcaceae | 24.7 ± 4.5 | 30.4 ± 6.4 | - | 18.2 ± 3.6b | ↓ | 26.7 ± 4.0 | - | 21.1 ± 4.0 | - | < 0.05 | |
Proteobacteria (%) | |||||||||||
Desulfovibrionaceae | 1.94 ± 0.39 | 1.68 ± 0.32 | - | 3.06 ± 0.64 | - | 3.64 ± 0.95ab | - | 1.30 ± 0.31d | - | < 0.01 | |
Enterobacteriaceae | 0.24 ± 0.04 | 14.38 ± 5.93a | ↑ | 6.28 ± 2.59a | - | 5.99 ± 3.80a | - | 0.55 ± 0.16bcd | - | < 0.01 | |
Escherichia Shigella | 0.05 ± 0.03 | 14.08 ± 5.83a | ↑ | 6.02 ± 2.53a | - | 5.47 ± 3.46bc | - | 0.30 ± 0.11bc | - | < 0.01 | |
Pseudomonadaceae | 0.11 ± 0.04 | 0.06 ± 0.01 | - | 0.06 ± 0.02 | - | 0.18 ± 0.09 | - | 0.21 ± 0.12 | - | NS | |
Sutterellaceae | 0.05 ± 0.40 | 1.02 ± 0.42 | - | 2.87 ± 0.87 | - | 4.97 ± 1.93 | - | 7.39 ± 2.97 | - | NS | |
Rhodospirillaceae | 0.14 ± 0.05 | 0.51 ± 0.26a | ↑ | 0.02 ± 0.02b | ↓ | 0.00 ± 0.00b | - | 0.04 ± 0.02b | - | < 0.001 | |
Tenericutes (%) | |||||||||||
Anaeroplasmataceae | 0.23 ± 0.13 | 0.00 ± 0.00a | ↓ | 0.00 ± 0.00a | - | 0.01 ± 0.01a | - | 0.00 ± 0.00a | ↓ | < 0.01 | |
Total SCFA (µmol/g) | 12.4 ± 1.63 | 1.13 ± 0.78a | ↓ | 0.06 ± 0.04a | - | 0.14 ± 0.07a | - | 4.88 ± 1.08abcd | ↓ | < 0.001 | |
Acetate | 8.95 ± 1.16 | 0.94 ± 0.67a | ↓ | 0.04 ± 0.04ab | ↓ | 0.14 ± 0.07ab | - | 3.40 ± 0.71abcd | ↓ | < 0.001 | |
Propionate | 1.93 ± 0.29 | 0.04 ± 0.04a | ↓ | 0.00 ± 0.00a | - | 0.00 ± 0.00a | - | 0.90 ± 0.26abcd | ↓ | < 0.001 | |
Butyrate | 1.51 ± 0.22 | 0.15 ± 0.11a | ↓ | 0.01 ± 0.01a | - | 0.00 ± 0.00a | - | 0.59 ± 0.13abcd | ↓ | < 0.001 |
- Citation: Vidal-Lletjós S, Andriamihaja M, Blais A, Grauso M, Lepage P, Davila AM, Gaudichon C, Leclerc M, Blachier F, Lan A. Mucosal healing progression after acute colitis in mice. World J Gastroenterol 2019; 25(27): 3572-3589
- URL: https://www.wjgnet.com/1007-9327/full/v25/i27/3572.htm
- DOI: https://dx.doi.org/10.3748/wjg.v25.i27.3572