Ischemia/Reperfusion-Induced Low Reactivity of the Rat Superior Mesenteric Vascular Bed is Associated With Expression of Nitric Oxide Synthases

Abstract

Our objective was to investigate the mRNA and protein expressions of eNOS and iNOS in the mesenteric vascular bed after ischemia and reperfusion of the rat superior mesenteric artery (SMA) and the role of nitric oxide (NO) in the response of the vascular bed to vasoconstrictors following reperfusion of the SMA. Methods Real-time polymerase chain reaction and immunohistochemistry were used to monitor the mRNA and protein expression of eNOS and iNOS after I/R challenge to the rat SMA. Ischemia was induced by clamping the SMA for 40 minutes, after which the flow was restored and the vessels were reperfused for 300 minutes. Blood samples were collected for assays of lactic dehydrogenase, tumor necrosis factor (TNF), hydroxyl radical, and NO. After ischemia/reperfusion, the vascular beds were separated for analysis of the expression of eNOS and iNOS. The SMA with its associated intestinal tissue was isolated and perfused in vitro with Tyrode’s solution ( N = 8) then challenged with phenylephrine. Results Reperfusion of the SMA induced an increase in blood concentrations of lactic dehydrogenase ( P < .001; N = 8), hydroxyl radical ( P < .05), TNF ( P < .001), and NO ( P < .05). ENOS and iNOS mRNA expression increased 1.3 ± 0.1-fold and 19.6 ± 3.5-fold, respectively when compared to the sham-operated group. Protein expression increased 1.9 ± 0.4-fold and 12.6 ± 3.1-fold, respectively, after reperfusion (N = 3) when compared with sham-treated rats. In vitro challenge showed that administration of phenylephrine (10 −8 ∼ 10 −4 nmol) produced vasoconstriction in a dose-related manner. Maximum contractile responses to phenylephrine were attenuated in reperfused SMA. Addition of the NOS inhibitor N G-nitro-L-arginine (L-NNA, 10 −4 M) resulted in full recovery of the response to phenylephrine. Conclusions Ischemia/reperfusion of the SMA results in a decrease in vascular reactivity of the mesenteric vessels that is dependent on NOS expression by the intestinal vascular bed.