Intestinal ischemia-reperfusion of macaques triggers a strong innate immune response

Table 1 Sequences of primers and polymerase chain reaction products

mRNA	Sense	Antisense	Size (bp)
TLR4	TGCAATGGATCAAGGACCAGAGGC	GTGCTGGGACACCACAACAATCACC	449
TLR2	GGCCAGCAAATTACCTGTGTG	CTGAGCCTCGTCCATGGGCCACTCC	637
MD2	GAAGCTCAGAAGCAGTATTGGGTC	GGTTGGTGTAGGATGACAAACTCC	422
IL-1β	AAACAGATGAAGTGGTCCTTCCAGG	TGGAGAACACCACTTGTTGCTCCA	388
TNF-α	AGGGCTCCAGGCGGTGCTTG	TGGTAGGAGACGGCGATGCGG	418
β-actin	CACCACACCTTCTACAATGAGC	GTGATCTCCTTCTGCATCCTGT	695

TLR: Toll-like receptor; IL: Interleukin; TNF: Tumor necrosis factor.

Table 2 Inflammatory lesion scores in two groups

Group	M/F	+	++	+++
Control	3/2	42	8	0
IIR	3/2	6a	30a	16a

^aP < 0.05 vs control group. M: Male; F: Female.

Table 3 Quantitative comparison of TLR4, TLR2, MD2, and NF-κB p65 protein in intestinal mucosa (IOD value)

Group	TLR4	TLR2	MD2	NF-κB p65	IFN-γ
Control	0.08 ± 0.03	0.07 ± 0.04	0.04 ± 0.02	0.19 ± 0.06	0.14 ± 0.05
IIR	0.22 ± 0.04a	0.13 ± 0.04a	0.16 ± 0.06a	0.65 ± 0.12a	0.38 ± 0.10a

n = 5 in each group,

^aP < 0.05 vs control group. There was a positive correlation between TLR4, TLR2 and MD2 and the expression of NF-κB p65 respectively (r = 0.870, P < 0.01; r = 0.720, P < 0.05). TLR: Toll-like receptor; NF-κB: Nuclear factor-κB; IFN-γ: interferon-γ; IIR: Intestinal ischemia-reperfusion.

Table 4 mRNA expression of TLRs, MD2 and inflammatory mediators in intestinal mucosa (IOD value)

Group	TLR4	TLR2	MD2	TNF-α	IL-1β
Control	0.5 ± 0.04	0.31 ± 0.05	0.77 ± 0.05	0.35 ± 0.08	0.18 ± 0.04
IIR	1.39 ± 0.06a	1.52 ± 0.15a	1.94 ± 0.12a	1.48 ± 0.15a	0.66 ± 0.08a

n = 5 in each group,

^aP < 0.05 vs control group. Toll-like receptor 4 (TLR4) was positively correlated with the expression of interleukin (IL)-1β (r = 0.964; P < 0.01), MD2 was positively correlated with the expression of IL-1β (r = 0.951; P < 0.01), TLR4 was positively correlated with the expression of tumor necrosis factor (TNF)-α (r = 0.951; P < 0.01) and MD2 was positively correlated with the expression of TNF-α (r = 0.955; P < 0.01). IIR: Intestinal ischemia-reperfusion.

Table 5 Inflammatory mediators in the small intestine mucosa and peripheral plasma

Group	IL-1β		IL-6		TNF-α
	Plasma	Ileum	Plasma	Ileum	Plasma	Ileum
	(pg/mL)	(pg/g protein)	(pg/mL)	(pg/g protein)	(pg/mL)	(pg/g protein)
Control	29.5 ± 7.3	76.8 ± 20.5	19.8 ± 8.2	663.4 ± 186.9	5.6 ± 1.7	49.0 ± 9.4
IIR	86.3 ± 15.2b	273.4. ± 44.7b	1129 ± 248.3b	1636 ± 168.0b	77.8 ± 16.2b	205.5 ± 30.7b

n = 5 in each group;

^bP < 0.01 vs control group. TNF-α: Tumor necrosis factor-α; IL: Interleukin; IIR: Intestinal ischemia-reperfusion.