Endothelin-1 in Human Intestine Resected for Necrotizing Enterocolitis

Objectives

We asked if the tissue concentration of the potent vasoconstrictor endothelin-1 (ET-1) is greater in areas of human preterm intestine that demonstrate histologic evidence of necrotizing enterocolitis (NEC) when compared with relatively healthy areas within the same resection specimen. We then evaluated if ET-1 participates in hemodynamic regulation within intestinal subserosal arterioles harvested from portions of human preterm intestine that demonstrate NEC.

Study design

Human preterm intestine resected for NEC was divided into three zones based on proximity to the perforation (zone 1 most proximal, zone 3 most distal). Histologic evidence of NEC was determined in each zone (normal=0, advanced necrosis=6). The tissue concentration of ET-1 was determined by enzyme-linked immunosorbent assay within intestinal homogenates prepared from each zone. Arteriolar hemodynamics were determined in vitro on subserosal arterioles harvested from different zones. Arteriolar flow rate, diameter, and resistance were determined at pressure gradients (ΔP) of 20 and 40 mmHg under control conditions and again after blockade of endothelin ET_A receptors with BQ610 (10⁻⁹mol/L).

Results

The tissue concentration of ET-1 (pg/mg protein) and histologic score in the three zones were: zone 1: 84 ± 14, 5.5 ± 0.3; zone 2: 99 ± 12, 4.7 ± 0.4, and zone 3: 33 ± 9, 0.8 ± 0.6, respectively (M ± SD, n=10 resection specimens, P <.05, zone 3 vs zones 1 and 2). Zone 2 arterioles demonstrated significantly lower flow rate and diameter and increased resistance under control conditions than zone 3 arterioles when ΔP was either 20 or 40 mmHg (n=7, P <.05). Treatment with BQ610 had no effect on zone 3 arterioles but significantly vasodilated zone 2 arterioles, increasing flow rate and vessel diameter, and decreasing vascular resistance (n=7, P <.05).

Conclusions

The tissue concentration of ET-1 is greater in human preterm intestine that demonstrates histologic evidence of NEC. Arterioles harvested from intestine exhibiting histologic evidence of NEC demonstrate vasoconstriction when compared with arterioles from relatively healthy intestine in the same resection specimen. This vasoconstriction was reversed by blockade of endothelin ET_A receptors.

ANOVA, Analysis of variance, ET-1, Endothelin-1, NEC, Necrotizing enterocolitis, PAF, Platelet activating factor, ΔP, Pressure gradient