Redox signaling of the arteriolar myogenic response.

Arteriolar vascular smooth muscle cells (VSMCs) are mechanosensitive, constricting to elevations in transmural pressure (P(TM)). The goal of the present study was to determine using mouse isolated tail arterioles and arteries whether oxidant signaling regulates this myogenic response. In response to P(TM) elevation, VSMCs of arterioles but not arteries generated constriction and increased reactive oxygen species (ROS) activity (using the H(2)O(2)-sensitive probe dichlorodihydrofluorescein). Arterioles had increased expression of NADPH oxidase components compared with arteries. Inhibition of NADPH oxidase, using mice with targeted impairment of enzyme components (p47(phox) or rac1) or diphenyleneiodonium, prevented the pressure-induced generation of ROS. When ROS activity was inhibited, either by inhibiting NADPH oxidase or with N-acetylcysteine, the myogenic constriction was abolished. The myogenic constriction was also inhibited by catalase, which inactivates H(2)O(2), but was unaffected by a cell-permeant mimic of superoxide dismutase (MnTMPyP). alpha(1)-Adrenergic constriction was not associated with altered ROS activity and was not affected by inhibition of NADPH oxidase or ROS. Exogenous H(2)O(2) constricted VSMCs of arterioles but not arteries. Thus, NADPH oxidase and ROS, in particular H(2)O(2), contribute to the myogenic response of arteriolar VSMCs.

Determinants of terminal mesenteric artery resistance during the first postnatal month.

Experiments were conducted to delineate the vascular effector systems that contribute to setting mesenteric vascular tone in swine during the first postnatal month. Terminal mesenteric arteries (TMA), which function as resistance vessels, were studied in vitro with a microvascular perfusion system allowing independent pressure and flow manipulation. When pressure was varied 0-100 mmHg in the absence of flow, TMA from 1-day-old animals demonstrated myogenic vasoconstriction, whereas TMA from 40-day-old animals did not. In 1- but not 40-day-old TMA, the endothelin A (ET(A)) receptor antagonist BQ-610 shifted the pressure-diameter curve upward, whereas the ET(B) receptor antagonist BQ-788 and the L-arginine analog N(G)-monomethyl-L-arginine (L-NMMA) shifted the curve downward; in all instances, myogenic vasoconstriction was preserved. Flow eliminated myogenic vasoconstriction in 1-day-old TMA, i.e., diameter increased as a function of pressure. The effect of BQ-610 was lost under flow conditions; however, BQ-788 and N-acyl-L-Trp-3,5-bis-(trifluoromethyl) benzyl ester, an antagonist specific to the substance P neurokinin-1 (NK(1)) receptor, shifted the pressure-diameter curve downward in the presence of flow, whereas L-NMMA restored myogenic vasoconstriction. Adding flow had no effect on the pressure-diameter relationship in 40-day-old TMA. Other blocking agents, including prazosin, losartan, indomethacin, and charybdotoxin, had no effect on the pressure-diameter relationship in either age group under flow or no-flow conditions. Constitutive production of nitric oxide (NO) and endothelin-1 participates in setting resistance in 1-day-old TMA, and important stimulants to NO production include flow and activation of ET(B) and NK(1) receptors. In contrast, 40-day-old TMA act as passive conduits in which the elastic properties of the vessel are the primary determinant of diameter.

Relationship between flow rate and NO production in postnatal mesenteric arteries.

We studied mesenteric arterial arcades from 3- and 35-day-old swine to determine the relationship between perfusate flow rate and release of nitric oxide (NO) into mesenteric effluent. Mesenteric arterial arcades were perfused under controlled-flow conditions with a peristaltic pump using warm oxygenated Krebs buffer. Basal rates of NO production were 43.6 +/- 4.2 vs. 12.1 +/- 2.5 nmol/min in 3- vs. 35-day-old mesentery during perfusion at in vivo flow rates (9 vs. 20 ml/min, respectively). Rate of NO production was directly related to flow rate over a wide range of flows (5-40 ml/min) in 3- but not 35-day-old mesentery. Both age groups demonstrated a brisk, albeit brief, increase in NO production in response to infusion of NO-dependent vasodilator substance P (10(-8) M/min). Tyrosine kinase inhibitor herbimycin A and L-arginine analog L-NMMA significantly attenuated flow-induced increase in NO production, and phosphatase inhibitor phenylarsine oxide increased magnitude of flow-induced increase in NO production in 3-day-olds. Removal of extracellular Ca(2+) and depletion of intracellular Ca(2+) stores (Ca(2+)-free Krebs with EGTA plus thapsigargin) had no effect on NO production in either group. Thus, basal rate of NO production is greater in mesenteric arterial arcades from 3- than from 35-day old swine, a direct relationship between flow rate and NO production rate is present in mesentery from 3- but not 35-day-olds, and phosphorylation events are necessary for this interaction to occur.

Age-dependent changes in the postnatal intestinal microcirculation.

Significant changes occur in intestinal hemodynamics during the transition from fetal to newborn life and then again during the first postnatal month. Most importantly, basal vascular resistance substantially decreases following birth. It then decreases further between postnatal days 1 and 3, plateaus, and then begins a slow, progressive increase between postnatal days 12 and 30. The basal rate of intestinal blood flow mirrors the changes in vascular resistance in an inverse manner. The postnatal changes in vascular resistance appear to be mediated, in large part, by an increase in the constitutive and stimulated production of nitric oxide. Most importantly, the diameter of terminal mesenteric arteries (150-300 microm diameter) in newborn (i.e., 1 day old) swine is determined by three intrinsic vascular control systems: endothelial production of nitric oxide and endothelin, and the inherent myogenic response of vascular smooth muscle. In contrast, these vessels in older subjects (i.e., 35 days old) are primarily passive in nature and fail to demonstrate significant diameter change in response to blockade of endogenous nitric oxide production or endothelin receptors, or applied perturbations of pressure or flow rate. The circulatory physiology of the perinatal and newborn intestine is exceptional when compared to the adult condition inasmuch as several hemodynamic variables change quite dramatically between fetal and neonatal life and during the first postnatal month. The unique hemodynamic conditions that characterize the perinatal and newborn intestine appear to be part of the overall physiological transition that occurs as the fetus, once born, replaces the placenta with his gastrointestinal tract to obtain nutrition. The goal of this review is to describe the circulatory physiology of the perinatal and newborn intestine, with a particular emphasis on those portions of the intestinal microcirculation that have thus far been studied. First, however, it is important to discuss the age-dependent changes that occur within the intestinal circulation during perinatal and early newborn life.

Role of endothelin-1 in regulation of the postnatal intestinal circulation.

Newborn intestine is uniquely prone to vasoconstriction in response to a wide variety of perturbations. To test the hypothesis that endothelin (ET)-1 is an important factor in this process, we determined the effects of exogenous ET-1 administration and blockade of endogenous ET-1 in vivo and in vitro in 3- and 35-day-old swine. Intramesenteric artery administration of exogenous ET-1 to vascularly isolated in vivo gut loops (10(-9) M/kg bolus) caused vasoconstriction and reduced gut O(2) uptake similarly in these age groups. Selective blockade of ET(A) or ET(B) receptors with BQ-610 or BQ-788, respectively, in vascularly isolated in vivo gut loops had no effect on gut vascular resistance or O(2) uptake in either age group; within in vitro gut loops, BQ-610 significantly increased vasoconstriction when perfusion pressure was reduced below baseline, but only in 3-day-old animals; i.e., it impaired the autoregulatory response to perfusion pressure reduction. Exogenous ET-1 significantly decreased capillary perfusion within in vitro gut loops, as evidenced by a decrease in capillary filtration coefficient, but only in 3-day-old animals; furthermore, blockade of endogenous ET-1 activity with BQ-610 significantly enhanced capillary filtration coefficient in 3-day-old animals and increased O(2) extraction ratio. ET-1 did not depress intestinal metabolic rate, as evidenced by its effect on the O(2) uptake-blood flow relationship; it did compromise tissue oxygenation because of its effects on intestinal O(2) transport. ET-1 concentration in mesenteric venous effluent exceeded arterial concentration, but only in 3-day-old intestine, suggesting production of ET-1 by newborn intestine. We conclude that ET-1 exerts an age-dependent effect on intestinal hemodynamics in postnatal intestine, having a greater impact in 3- than in 35-day-old intestine.

Heparin-binding epidermal growth factor-like growth factor protects rat intestine from ischemia/reperfusion injury.

BACKGROUND: We have shown previously that heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) is cytoprotective for intestinal epithelial cells exposed to hypoxia in vitro. We now examine the effects of HB-EGF on the recovery of small intestine from ischemic injury in vivo. METHODS: Segmental intestinal ischemia of 60-min duration was produced in adult rats by occlusion of a first-order branch of the superior mesenteric artery. Recombinant HB-EGF (100 microg) was injected intraluminally into the proximal small bowel after 45 min of ischemia in experimental animals, and buffered saline was injected in control animals. Animals were sacrificed after 48 h, and the affected bowel was resected, processed, and examined microscopically, with histologic grading of the ischemic injury. Additional animals were allowed to recover for up to 1 month to evaluate mortality differences. RESULTS: Intraluminal administration of HB-EGF resulted in significantly decreased extent and severity of ischemia/reperfusion injury, with significantly decreased grade of injury in the HB-EGF-treated compared with nontreated animals (average injury grade 0.66 compared with 2.44, respectively). Moreover, the mortality rate was significantly lower in the HB-EGF-treated animals compared with nontreated animals (0% vs 25%, respectively). HB-EGF-treated animals had increased weight gain in the postischemia recovery period. CONCLUSIONS: We conclude that HB-EGF, given intraluminally, reduces both the amount and the severity of ischemia/reperfusion injury in the small bowel, reduces the mortality associated with intestinal ischemia, and may enhance intestinal recovery. The in vitro and in vivo cytoprotective effects of this growth factor suggest that it may, in the future, be clinically useful in treating patients with intestinal ischemia.

Effects of sustained low-flow perfusion on the response to vasoconstrictor agents in postnatal intestine.

This laboratory has previously reported that sustained reduction of blood flow in newborn intestine causes a triphasic increase in vascular resistance that occurs over 3-4 h and that these changes are mediated, in part, by loss of endothelial nitric oxide (NO) production. This study examines the effects of exposure to sustained low-flow perfusion on the subsequent response to three contractile agonists: ANG II, norepinephrine (NE), and endothelin-1 (ET-1). Gut loops from 3- and 35-day-old swine were exposed to low-flow conditions in vivo (i.e., reduction of flow to approximately 50% of baseline) for 30 min or 5 h. Thereafter, they were removed to an extracorporeal perfusion circuit for in vitro hemodynamic assessment; alternatively, the mesenteric artery perfusing the gut loop was removed and cut into rings for assessment of isometric tension development. Gut loops from 3-day-old subjects exposed to low-flow conditions demonstrated significantly increased contractile responses to ANG II, NE, and ET-1; also, mesenteric artery rings from these gut loops demonstrated a significant reduction of the ED50 for all three agonists. Similar changes were not observed in intestine or mesenteric artery rings from older subjects. Sustained blockade of endogenous NO synthesis with NG-monomethyl- L-arginine duplicated the effects of exposure to sustained low-flow perfusion. It appears that sustained reduction of blood flow in newborn intestine decreases constitutive NO production, which in turn causes a generalized enhancement of the contractile efficacy of ANG II, NE, and ET-1.

Effects of systemic hypotension on postnatal intestinal circulation: role of angiotensin.

Systemic hypotension causes a greater degree of vasoconstriction in intestine from 3- than from 35-day-old postnatal swine. To determine the basis for this age-dependent difference, systemic hypotension (pressure reduction to approximately 50% of baseline) was induced by creating pericardial tamponade in postnatal swine instrumented to allow measurement of intestinal hemodynamics and oxygenation in vivo. Hypotension caused gut vascular resistance to increase 77 +/- 6% in 3-day-old subjects but only 18 +/- 3% in 35-day-old subjects. Prior blockade of alpha1-receptors with phentolamine, vasopressin receptors with [d(CH2)5,D-Phe2,Ile4,Ala9-NH2]AVP, or surgical denervation of the gut loop had no effect on hypotension-induced gut vasoconstriction. Losartan, which blocks angiotensin AT1 receptors, significantly attenuated hypotension-induced gut vasoconstriction in both age groups. BQ-610, which blocks endothelin ETA receptors, also limited the magnitude of vasoconstriction but only in younger subjects. This effect may have been consequent to an interaction between endothelin and angiotensin, inasmuch as a subpressor concentration of endothelin increased the contractile response to angiotensin in mesenteric artery rings. The substantial rise in 3-day-old gut vascular resistance was partly consequent to a locally mediated vasoconstriction that occurred in response to pressure and/or flow reduction during hypotension, as evidenced by the significant attenuation of this constriction when blood flow was held constant by controlled-flow perfusion to the gut loop during hypotension. Intestinal O2 uptake was compromised to a significantly greater degree in 3- than in 35-day-old subjects during hypotension. This difference was primarily due to the inability of younger intestine to increase O2 extraction in the face of reduced blood flow and may be mediated, in part, by an effect of angiotensin II on intestinal capillary perfusion.

Effects of sustained flow reduction on postnatal intestinal circulation.

Studies were conducted to determine the effect of mechanically induced sustained flow reduction on intestinal hemodynamics and oxygenation in 3- and 35-day-old swine. In vitro gut loops were perfused under controlled-pressure conditions from an oxygenated blood reservoir at age-appropriate perfusion pressures; pressure was rapidly reduced to a level that lowered flow rate to approximately 50% of its baseline value, and pressure was then kept at that level for 2 h. In 3-day-old intestine, vascular resistance (Ri) increased by 20% immediately after pressure and flow reduction but then stabilized for 3-4 min; thereafter, flow began to decrease despite maintenance of perfusion pressure, so that Ri increased an additional 15% by 30 min after flow reduction. Flow continued to diminish over the next 90 min, though at much slower rate. Intestine from 35-day-old swine demonstrated an immediate increase in Ri after pressure and flow reduction, but thereafter Ri increased very little. The protocol was repeated within in vitro gut loops perfused under controlled-flow conditions, and within autoperfused, innervated gut loops developed in vivo and similar observations were made in both preparations. In 3-day-old intestine, pretreatment with the L-arginine analog Nomega-monomethyl-L-arginine (10(-4) M) had no effect on the immediate rise in resistance occurring in the first 1 min but substantially attenuated the subsequent slow, progressive rise noted thereafter. Pretreatment with the angiotensin 1A receptor antagonist losartan (2 x 10(-6) M) had no effect on hemodynamic changes during the first 60 min after mechanical perfusion pressure reduction but attenuated the very slight increase in resistance noted during the final 60 min of the protocol. The postnatal intestinal circulation demonstrates progressive vasoconstriction when its flow rate is mechanically reduced in a sustained manner, and this effect is age specific, occurring in 3- but not 35-day-old swine. These changes in gut vascular resistance may be consequent to loss of nitric oxide production and/or local production of angiotensin.

Postnatal changes in gut hemodynamics: a possible role for substance P.

Studies were conducted in young postnatal swine to determine if substance P (SP) participates in the regulation of postnatal intestinal hemodynamics and oxygenation. SP was present in homogenates of whole intestine from postnatal swine in an age-dependent manner as follows: 1 day old and never fed, 126 +/- 35; 3 days old and fasted, 148 +/- 30; and 14 days old, 51 +/- 10 pg/mg protein (P < 0.01, 14- vs. 1- or 3-day-olds). Phenylephrine-precontracted rings of mesenteric artery from 3-day-old subjects mounted for tension recording within buffer-filled myographs demonstrated brisk relaxation in response to SP (EC50, 2 x 10(-10) M). This relaxation was eliminated by mechanical removal of the endothelium or blockade with the L-arginine analog NG-monomethyl-L-arginine (L-NMMA) and was significantly attenuated by pretreatment with N-acyl-L-Trp-3,5-bis-(trifluoromethyl) benzyl ester (NATB), a highly selective NK-1 receptor antagonist (pA2 5 x 10(-10) M). Infusion of exogenous SP into the mesenteric artery of innervated in vivo gut loops reduced intestinal vascular resistance 35% and increased tissue oxygen uptake 40% in both 3- and 14-day-old subjects. By contrast, blockade of the NK-1 receptor for SP with NATB increased intestinal vascular resistance 19% in 3-day-old subjects but only 5% in 14-day-old subjects (P < 0.01). SP-induced changes in gut vascular resistance were significantly attenuated by prior coinfusion of NATB or L-NMMA, indicating that the peptide exerted this vascular effect via theNK-1 receptor, which is linked to endothelial cell nitric oxide synthase. Both NATB and L-NMMA attenuated flow-induced dilation within pump-perfused in vitro gut loops from 3-day-old subjects. SP appears to participate in the regulation of the newborn intestinal circulation, especially during the first days after birth.

Segmental intestinal ischemia: an improved method of producing small bowel injury.

Previous animal models of intestinal ischemia-reperfusion have been successful in causing considerable mucosal damage, cellular destruction and sepsis. However, this often results in the death of the animal, making it impossible to examine the effects of modulators of the ischemic event. The sequence of morphologic and physiologic changes in the bowel from such injuries continues to be an area of intense examination. We have studied these changes by producing segmental intestinal ischemia in vivo in a rat model. By occluding a first-order branch of the superior mesenteric artery (SMA) and by selectively ligating terminal collateral branches, reproducible segmental intestinal ischemia was achieved. Bowel damage ranged from alterations in the villus structure to frank hemorrhagic necrosis of the intestinal wall. This model allows the study of hypoperfusion injury to the small intestine without total SMA occlusion, thus reducing the overall mortality.

Pressure and flow characteristics of terminal mesenteric arteries in postnatal intestine.

The responses of buffer-perfused terminal mesenteric arteries from 3- and 35-day-old swine to manipulation of intravascular pressure and flow rate were determined. Under in vivo conditions, these vessels demonstrated age-dependent differences in resting diameter (182 vs. 301 microns in 3- vs. 35-day-old swine). A proximal-to-distal pressure gradient was present in vessels from both age groups (delta 13 vs. delta 16 mmHg in 3- vs. 35-day-old swine), suggesting their functional role as resistance vessels. Vessels were mounted within an in vitro perfusion apparatus that allowed independent regulation of inflow and outflow pressure. Vessels from both age groups demonstrated the development of active tone in response to an incremental rise in pressure, applied in the absence of flow. However, myogenic vasoconstriction was only observed in younger arterioles. Similarly, both groups demonstrated dilation in response to a flow stimulus generated in the absence of a net change in intravascular pressure, although the magnitude of this response was significantly greater in younger vessels (+27 vs. +7% in 3- vs. 35-day-old swine). The dilatory response to flow was eliminated by NG-monomethyl-L-arginine (10(-4)M) but restored by coadministration of L-arginine (10(-3)M). Myogenic vasoconstriction was overridden by flow-mediated dilation in terminal mesenteric arteries from 3- but not 35-day-old swine after concomitant application of pressure and flow stimuli. We conclude that the hemodynamic characteristics of terminal mesenteric arteries are age dependent in postnatal swine.

The effects of ischemia-reperfusion on endothelial cell function in postnatal intestine.

The goal of these experiments was to determine whether the perturbation of ischemia-reperfusion has an age-dependent effect on subsequent endothelial cell production of nitric oxide. Three- and 35-d-old swine in the experimental group were exposed to 1-h partial ischemia (90% flow reduction) and 2-h reperfusion in vivo by creation and then removal of a mesenteric artery coarctation. Control subjects underwent exposure of the mesenteric artery only. After reperfusion, gut vascular resistance had increased 44 +/- 6% in 3-d-old, but had decreased 41 +/- 4% in 35-d-old subjects. At the completion of the in vivo portion of the protocol mesenteric artery was removed, and nitric oxide production was estimated in vitro, by measuring cGMP production by vessel segments or by measuring relaxation of phenylephrine-precontracted rings, both after stimulation of nitric oxide production by substance P or the calcium ionophore A23187. Compared with control, mesenteric artery segments from 3-d-old subjects demonstrated reductions in basal, substance P-stimulated (10(-8) M) and A23187-stimulated (10(-7) M) cGMP accumulation of 50 +/- 7%, 66 +/- 6% and 78 +/- 7%. Mesenteric artery segments from 35-d-old subjects demonstrated increases in basal, substance P-stimulated, or A23187-stimulated cGMP accumulations of 114 +/- 14%, 92 +/- 8%, or 78 +/- 9%. Compared with control, I/R rings from 3-d-old subjects demonstrated reductions in substance P-induced (10(-8) M) or A23187-induced (10(-7) M) relaxations of 56 +/- 7% or 30 +/- 7%. In contrast, 35-d-old ischemia-reperfusion rings demonstrated increases in substance P- or A23187-induced relaxation of 36 +/- 8% or 98 +/- 11%. It is concluded that ischemia-reperfusion has an age-dependent effect on endothelial production of NO within in vitro postnatal mesenteric artery and that these changes mirror the effects of ischemia-reperfusion on gut vascular resistance in vivo.

The effects of systemic hypotension on postnatal intestinal hemodynamics and oxygenation.

This study investigated the effect of acute systemic hypotension, induced by pericardial tamponade, on intestinal hemodynamics and oxygenation in 3- and 35-d-old swine. To delineate intrinsic versus systemic responses, the effects of tamponade were compared with those noted after isolated pressure reduction to an innervated in vivo gut loop, achieved by local arterial constriction. Younger subjects demonstrated 16 and 58% increases in vascular resistance during constriction and tamponade, respectively, whereas older subjects exhibited an increase in resistance (27%) only during tamponade. Intestinal oxygen uptake decreased approximately 30% during both constriction and tamponade in the younger group, despite the greater effect of tamponade on gut perfusion. Older subjects exhibited no change in gut oxygenation during either perturbation. In a separate series of experiments, vasopressin, phenylephrine, and angiotensin II, each an element of the systemic pressor response, were infused into in vitro gut loops from 3- and 35-d-old swine. Vasopressin caused sustained vasoconstriction in both age groups; however, phenylephrine and angiotensin II caused greater sustained increases in intestinal vascular resistance in 3- than in 35-d-old intestine. We conclude that systemic hypotension compromises intestinal perfusion to a greater extent in younger subjects, although this effect is not associated with overwhelming tissue hypoxia. The pronounced rise in gut vascular resistance in 3-d-old intestine may reflect the additive effects of intrinsic and systemic vasoactive forces engaged during acute systemic hypotension.

Flow-induced dilation in newborn intestine.

The goal of these experiments was to determine the presence and mechanistic basis of flow-induced dilation in mesenteric artery from 3-d-old swine. In the first experiment, in vitro gut loops were perfused from a blood-filled reservoir under controlled-flow conditions, and flow was progressively increased from approximately 40% to approximately 170% in six increments by manipulation of pump speed. Under control conditions, vascular resistance significantly decreased after each step increase in flow rate. NG-Monomethyl-L-arginine (LNMMA; 10(-4) M), an arginine analog that blocks nitric oxide production, eliminated this flow-induced dilation, but only for the step increases in flow at rates above the baseline flow rate. For step increases below the baseline rate, LNMMA caused a simple parallel shift of the resistance-flow rate curve upward. Phenylephrine (10(-6) M), an alpha 1-agonist which has no effect on nitric oxide production or half-life, did not eliminate flow-induced dilation, but instead caused a simple parallel shift of the resistance-flow rate curve upward across the entire range of flows studied. In the second experiment, a 3-cm segment of mesenteric artery was perfused with Krebs buffer at two flow rates: 10 and 25 mL/min. The effluent from the mesenteric artery segment was suffused onto a deendothelialized, phenylephrine-precontracted ring of swine carotid artery; relaxation of this bioassay vessel served as an index of release of relaxing factors from the mesenteric artery segment. Under control conditions, increase in the mesenteric artery flow rate caused a 60% relaxation of the bioassay vessel. This effect was eliminated by the addition of LNMMA to the buffer (10(-4) M), but not by the addition of indomethacin (10(-5) M). Flow-induced dilation occurs in the mesenteric artery of 3-d-old swine. This vascular phenomenon appears to be mediated by nitric oxide, but only at flows above the baseline flow rate. The mechanism(s) responsible for this phenomenon at lower flow rates is not clear.

Role of nitric oxide in regulation of vascular resistance in postnatal intestine.

Studies were conducted to determine whether endothelial production of nitric oxide (NO) participates in the regulation of vascular resistance in postnatal swine intestine. In vivo, intra-arterial infusion of the arginine analogue NG-monomethyl-L-arginine (L-NMMA, 10(-4) M) increased intestinal vascular resistance 34% in 3-day-old animals and 9% in 35-day-old animals (P < 0.01); similar findings were noted during infusion of 10(-3) M L-NMMA. Mechanical augmentation of gut flow rate induced intestinal vasodilation in both age groups; L-NMMA eliminated this flow-induced dilation in intestine of 3- but not 35-day-old animals. In vitro, precontracted mesenteric artery rings from both age groups relaxed to a similar extent in response to the endothelium-independent nitrovasodilator sodium nitroprusside (SNP) and the calcium ionophore A-23187; the effect of A-23187, but not SNP, was eliminated by mechanical disruption of the endothelium. Acetylcholine (ACh) and substance P (SP), agents with vascular effects that are secondary to receptor-mediated activation of NO, caused greater relaxation of rings from younger than from older animals, and this effect was attenuated by L-NMMA or methylene blue. Unstimulated accumulation of guanosine 3',5'-cyclic monophosphate (cGMP) occurred to a similar extent in vessel segments from both groups. ACh and SP increased cGMP accumulation in segments from 3- but not from 35-day-old animals. We conclude that the NO-cGMP axis participates to a greater extent in regulation of intestinal vascular resistance in 3- than in 35-day-old swine.

The role of the circulation in the pathogenesis of necrotizing enterocolitis.

Early descriptions of necrotizing enterocolitis (NEC) proposed gut ischemia as a primary factor in pathogenesis. Subsequent laboratory and clinical investigations have failed to support these theories, relegating most to positions of historical significance. Presently, the role of the circulation in the pathogenesis of NEC is unclear; however, several lines of ongoing investigation have begun to provide novel insight into the means by which circulatory aberration may initiate or contribute to the development of bowel necrosis.

Regulation of capillary exchange capacity in postnatal swine intestine.

To determine whether intrinsic regulation of capillary exchange capacity is age-dependent during early postnatal life, we measured the capillary filtration coefficient (Kf,c), an index of capillary exchange capacity, within isolated perfused intestine from 3- and 35-day-old swine at different arterial pressures. Kf,c was determined gravimetrically, by means of the flow-equilibration technique. During baseline perfusion at age-appropriate pressures, Kf,c averaged 0.203 +/- 0.020 vs. 0.361 +/- 0.040 ml.min-1 x mmHg-1 x 100 g in 35- vs. 3-day-old intestine. In older animals, a progressive rise in Kf,c was noted as arterial pressure was lowered: Kf,c increased 14, 35, and 98% above baseline when pressure was reduced 20, 40, and 60% below baseline, respectively. An inverse linear correlation between Kf,c and blood flow was demonstrated in this group. Kf,c also increased in 3-day-old intestine during progressive hypotension, but these changes were of less magnitude: Kf,c averaged 19, 17, and 23% above baseline when pressure was reduced 20, 40, and 60% below baseline, respectively. In contrast to the circumstance noted in older animals, the relationship between Kf,c and blood flow was not significant within 3-day-old intestine. Intraarterial infusion of 1 microgram/min of isoproterenol, an agent known to increase capillary exchange capacity, caused Kf,c to increase 80% within 3-day-old intestine group. We conclude that Kf,c increases to a greater extent in 35- than 3-day-old swine intestine in response to arterial pressure reduction. However, intestine from younger animals is capable of increasing capillary exchange capacity in response to stimuli other than arterial hypotension, as evidenced by its response to isoproterenol.

Effects of ischemia and reperfusion on intrinsic vascular regulation in the postnatal intestinal circulation.

The purpose of these experiments was to determine the effect of 1 h of total ischemia followed by 2 h of reperfusion on intrinsic vascular regulation within intestine from 3- and 35-d-old swine. Intrinsic vascular regulation was defined as the ability of in vitro segments of small intestine to bring about adjustments of blood flow and the arteriovenous O2 content difference across the intestinal segment of sufficient magnitude to preserve tissue O2 uptake in response to a 35% reduction in arterial perfusion pressure. This response was elicited before (control conditions) and after ischemia-reperfusion (post-I/R). In older subjects, the efficacy of blood flow regulation was attenuated post-I/R, insofar as blood flow fell in response to pressure reduction. However, this group demonstrated a rise in arteriovenous content difference after pressure reduction under control and post-I/R conditions that were of sufficient magnitude to preserve tissue O2 uptake. In younger subjects, blood flow regulation was absent under control conditions and post-I/R. The arteriovenous O2 content difference increased in response to pressure reduction under control conditions but failed to do so post-I/R; consequently, tissue oxygenation decreased in response to arterial pressure reduction post-I/R in 3-d-old intestine. We conclude that ischemia-reperfusion affects intrinsic vascular regulation in postnatal intestine and that this effect is age-dependent. Intestine from older subjects maintains the intrinsic capacity to preserve tissue oxygenation in response to a hypotensive challenge despite the insult of ischemia-reperfusion, whereas intestine from younger subjects does not maintain this capacity.

Effect of increased tissue oxygen uptake on autoregulation in postnatal intestine.

To determine whether the rate of tissue oxygen utilization affects pressure-flow autoregulation in an age-dependent fashion in postnatal swine intestine, in vitro gut loops from 3- and 35-day-old swine were subjected to a 35% step reduction in arterial pressure under control conditions, during intra-arterial infusion of 2,4-dinitrophenol (DNP), and 30 min after luminal instillation of predigested artificial swine milk. Autoregulation was quantitated by determining the effect of pressure reduction on vascular resistance, and also by calculating Gf, a flow-controlling gain factor that relates pressure and flow. DNP infusion increased oxygen uptake 77 and 58% in gut from 3- and 35-day-old swine, respectively, whereas feeding increased oxygen uptake approximately 50% in both groups. Under control conditions, arterial pressure reduction had no effect on vascular resistance in either group. During DNP infusion and 30 min after feeding, however, intestine from 35- but not from 3-day-old swine demonstrated significant vasodilation in response to pressure reduction. Gf averaged -0.06 +/- 0.11 vs. 0.21 +/- 0.08 (P < 0.05) before vs. DNP infusion, and 0.06 +/- 0.03 vs. 0.22 +/- 0.06 (P < 0.05) before vs. 30 min after feeding in intestine from 35-day-old swine; these increases in Gf indicate that the intensity of the pressure-flow response increased during experimental treatments. In contrast, Gf averaged -0.11 +/- 0.07 vs. -0.23 +/- 0.08 before vs. DNP infusion, and -0.23 +/- 0.06 vs. -0.23 +/- 0.09 before vs. 30 min after feeding in intestine from 3-day-old swine.(ABSTRACT TRUNCATED AT 250 WORDS)