Nitric oxide-releasing aspirin but not conventional aspirin improves healing of experimental colitis

Table 1 Primer sequence used in reverse transcription polymerase chain reaction determination of gene expression of proinflammatory factors

Gene		Primer
β-actin	Upstream	5’-TTG TAA CCA ACT GGG ACG ATA TGG-3’
	Downstream	5’-GAT CTT GAT CTT CAT GGT GCT AGG-3’
IL-1β	Upstream	5’-GCT ACC TAT GTC TTG CCC GT-3’
	Downstream	5’–GAC CAT TGC TGT TTC CTA GG-3’
TNF-α	Upstream	5’-TAC TGA ACT TCG GGG TGA TTG GTC C-3’
	Downstream	5’–CAG CCT TGT CCC TTG AAG AGA ACC-3’
COX-2	Upstream	5’–ACA ACA TTC CCT TCC TTC-3’
	Downstream	5’–CCT TAT TTC CTT TCA CAC C-3’
iNOS	Upstream	5’–CCA CAA TAG TAC AAT ACT ACT TGG–3’
	Downstream	5’–ACG AGG TGT TCA GCG TGC TCC ACG–3’

TNF: Tumor necrosis factor; COX: Cyclooxygenase; iNOS: Inducible form of nitric oxide synthase.

Table 2 Evaluation of related markers in trinitrobenzenesulfonic acid rats treated ig with non-selective and selective cyclooxygenase-1 and cyclooxygenase-2 inhibitors or nitric oxide-aspirin throughout the period of 10 d in 6-10 rats per group (mean ± SE)

Treatment	Weight (mg)	MPO (ng/mL)	IL1-β(pg/mL)	TNF-α(pg/mL)
Vehicle	1150 ± 23.1	51 ± 8.1	42 ± 3.4	4.8 ± 0.6
ASA (80 mg/kg)	1580 ± 41.4a	78 ± 9.3a	65 ± 5.1a	8.5 ± 0.95a
Indomethacin (5 mg/kg)	1450 ± 45.8a	65 ± 5.2a	58 ± 4.1a	7.8 ± 0.62a
SC-560 (5 mg/kg)	1310 ± 32.4a	52 ± 6.8a	53 ± 3.9a	7.5 ± 0.46a
Celecoxib (10 mg/kg)	1082 ± 14.9c	44 ± 4.3c	36 ± 2.2c	3.8 ± 0.33c
NO-ASA (80 mg/kg)	710 ± 12.3e	35 ± 3.2e	18 ± 1.5e	2.4 ± 0.21e

MPO: Myeloperoxidase; TNF: Tumor necrosis factor; ASA: Aspirin; SC-560: 5-​(4-​chlorophenyl)-​1-​(4-​methoxyphenyl)-​3-​(trifluoromethyl)-​1H-​pyrazole; NO: Nitric oxide.

^aP < 0.05 vs vehicle;

^cP < 0.05 vs vehicle, ASA, indomethacin or SC-560 groups;

^eP < 0.05 vs groups treated with non-selective and selective COX-1 or COX-2 inhibitors.

Table 3 The generation of prostaglandin E₂ in intact rats and those treated rats (mean ± SE)

Group	PGE2 generation in the colonic mucosa (pg/mg tissue weight)
Intact	182 ± 15.4
Vehicle	409 ± 23.2a
ASA (80 mg/kg per day)	130 ± 11.7c
Indomethacin (5 mg/kg per day)	124 ± 17.8c
SC-560 (5 mg/kg per day)	145 ± 15.6c
Celecoxib (10 mg/kg per day)	191 ± 18.3e
NO-ASA (80 mg/kg per day)	175 ± 17.3e

PGE₂: Prostaglandin E₂; ASA: Aspirin; SC-560: 5-​(4-​chlorophenyl)-​1-​(4-​methoxyphenyl)-​3-​(trifluoromethyl)-​1H-​pyrazole; NO: Nitric oxide.

^aP < 0.05 vs intact colonic mucosa;

^cP < 0.05 vs vehicle (control);

^eP < 0.05 vs ASA, indomethacin and SC-560 groups.