TLR4-HMGB1-, MyD88- and TRIF-dependent signaling in mouse intestinal ischemia/reperfusion injury

doi:10.3748/wjg.v21.i27.8314

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

Basic Study

World J Gastroenterol. Jul 21, 2015; 21(27): 8314-8325
Published online Jul 21, 2015. doi: 10.3748/wjg.v21.i27.8314

Table 1 Histologic grading of intestinal ischemia-reperfusion injury

Grade	Features
0	Normal,villus to crypt ratio 5-6:1, minimal number of lymphocytes and plasma cells, tall columnar surface epithelial cells
1	Epithelial cell degenerative changes (cuboidal,vacuolated) but intact, mild increase of lymphocytes and plasma cells in lamina propria
2	Decreased villus height, yielding villus to crypt ratio ≤ 1, epithelial cell necrosis or erosions, more chronic inflammation in lamina propria ± neutrophils, glandular dilatation
3	Villi effaced (flat surface), epithelial cell necrosis or erosions, pseudomembrane may appear on surface, glandular destruction, inflammation extending deep to muscle layer
4	Transmural changes (all of above plus change in muscle layer)

Table 2 Concentrations of inflammatory factors in serum (mean ± SD)

Group(n = 8)	p65 (pg/mL)	IL-6 (pg/mL)	TNF-α(pg/mL)
Sham	104.64 ± 11.89	50.02 ± 6.33	43.79 ± 4.18
Control	228.53 ± 24.85^a	104.91 ± 31.18^a	70.81 ± 6.97^a
Anti-HMGB1	145.00 ± 33.63^a^b	62.28 ± 6.73^a^b	52.76 ± 5.71^a^b
Anti-MyD88	191.12 ± 13.22^a^b^c	85.90 ± 17.37^a^b^c	63.19 ± 5.47^a^b^c
Anti-TRIF	183.73 ± 10.81^a^b^c	78.14 ± 7.38^a^b^c	59.70 ± 4.63^a^b^c

^aP < 0.05 vs sham group;

^bP < 0.05 vs control group;

^cP < 0.05 vs anti-HMGB1 group. HMGBI: High-mobility group protein 1; IL: Interleukin; MyD88: Myeloid differentiation gene 88; TNF-α: Tumor necrosis factor-α; TRIF: Translocating-chain-associating membrane protein.

Table 3 Levels of myeloperoxidase activity in the lung and liver (mean ± SD)

Group (n = 4)	Lung	Liver
Sham	0.30 ± 0.12	0.25 ± 0.03
Control	0.76 ± 0.06^a	0.68 ± 0.08^a
Anti-HMGB1	0.46 ± 0.06^a^b	0.40 ± 0.07^a^b
Anti-MyD88	0.63 ± 0.03^a^b^c	0.55 ± 0.03^a^b^c
Anti-TRIF	0.57 ± 0.06^a^b^c	0.50 ± 0.04^a^b^c

^aP < 0.05 vs sham group;

^bP < 0.05 vs control group;

^cP < 0.05 vs anti-HMGB1 group. HMGBI: High-mobility group protein 1; MyD88: Myeloid differentiation gene 88; TRIF: Translocating-chain-associating membrane protein.

Table 4 Lung injury score in all groups (mean ± SD)

Group (n = 6)	Alveolar congestion	Hemorrhage	Neutrophil infiltration	Alveolar wall thickness	Total score
Sham	1.18 ± 0.21	0.63 ± 0.27	0.23 ± 0.08	0.17 ± 0.08	2.22 ± 0.64
Control	3.82 ± 0.18	2.72 ± 0.18	1.85 ± 0.15	2.37 ± 0.23	10.75 ± 0.73^a
Anti-HMGB1	2.05 ± 0.12	1.02 ± 0.26	0.47 ± 0.10	0.55 ± 0.14	4.08 ± 0.60^a^b
Anti-MyD88	3.07 ± 0.16	2.13 ± 0.26	0.87 ± 0.18	1.08 ± 0.23	7.15 ± 0.77^a^b^c
Anti-TRIF	2.93 ± 0.16	1.92 ± 0.18	0.77 ± 0.14	1.02 ± 0.16	6.63 ± 0.62^a^b^c

^aP < 0.05 vs sham group;

^bP < 0.05 vs control group;

^cP < 0.05 vs anti-HMGB1 group. HMGBI: High-mobility group protein 1; MyD88: Myeloid differentiation gene 88; TRIF: Translocating-chain-associating membrane protein.

Table 5 Intestine injury score in all groups (mean ± SD)

Group (n = 6)	Jejunum	Ileum
Sham	0.17 ± 0.08	0.22 ± 0.12
Control	2.97 ± 0.16^a	3.15 ± 0.32^a
Anti-HMGB1	1.23 ± 0.23^a^b	1.32 ± 0.27^a^b
Anti-MyD88	2.17 ± 0.22^a^b^c	2.18 ± 0.29^a^b^c
Anti-TRIF	2.08 ± 0.17^a^b^c	2.05 ± 0.27^a^b^c

^aP < 0.05 vs sham group;

^bP < 0.05 vs control group;

^cP < 0.05 vs anti-HMGB1 group. HMGBI: High-mobility group protein 1; MyD88: Myeloid differentiation gene 88; TRIF: Translocating-chain-associating membrane protein.

Table 6 HMGB1and NF-κB mRNA expression in lungs from all groups (mean ± SD)

Group (n = 6)	HMGB1	NF-κB
Sham	1.04 ± 0.19	1.03 ± 0.21
Control	2.67 ± 0.30^a	2.04 ± 0.29^a
Anti-HMGB1	1.89 ± 0.18^a^b	1.42 ± 0.23^a^b
Anti-MyD88	2.35 ± 0.31^a^b^c	1.77 ± 0.18^a^b^c
Anti-TRIF	2.29 ± 0.28^a^b^c	1.70 ± 0.13^a^b^c

^aP < 0.05 vs sham group;

^bP < 0.05 vs control group;

^cP < 0.05 vs anti-HMGB1 group. HMGBI: High-mobility group protein 1; MyD88: Myeloid differentiation gene 88; NF-κB: Nuclear factor-κB; TRIF: Translocating-chain-associating membrane protein.

Table 7 HMGB1, NF-κB, MyD88, and TRIF mRNA expression in ileum from all groups (mean ± SD)

Group (n = 6)	HMGB1	NF-κB	MyD88	TRIF
Sham	1.04 ± 0.19	1.03 ± 0.21	1.01 ± 0.10	1.01 ± 0.11
Control	2.67 ± 0.30^a	2.04 ± 0.29^a	2.55 ± 0.16^a	3.05 ± 0.10^a
Anti-HMGB1	1.89 ± 0.18^a^b	1.42 ± 0.23^a^b	2.37 ± 0.08^a^b	2.90 ± 0.13^a^b
Anti-MyD88	2.35 ± 0.31^a^b^c	1.77 ± 0.18^a^b^c	2.30 ± 0.16^a^b	3.04 ± 0.13^a^c
Anti-TRIF	2.29 ± 0.28^a^b^c	1.70 ± 0.13^a^b^c	2.53 ± 0.07^a^c^d	2.85 ± 0.12^a^b^d

^aP < 0.05 vs sham group;

^bP < 0.05 vs control group;

^cP < 0.05 vs anti-HMGB1 group;

^dP < 0.05 vs anti-MyD88 group. HMGBI: High-mobility group protein 1; MyD88: Myeloid differentiation gene 88; NF-κB: Nuclear factor-κB; TRIF: Translocating-chain-associating membrane protein.

Citation: Wang J, He GZ, Wang YK, Zhu QK, Chen W, Guo T. TLR4-HMGB1-, MyD88- and TRIF-dependent signaling in mouse intestinal ischemia/reperfusion injury. World J Gastroenterol 2015; 21(27): 8314-8325
URL: https://www.wjgnet.com/1007-9327/full/v21/i27/8314.htm
DOI: https://dx.doi.org/10.3748/wjg.v21.i27.8314