Continuous monitoring of gastroduodenal mucosal hemodynamics in rats by laser-Doppler flowmetry and reflectance spectrophotometry

Gastroduodenal mucosal hemodynamics in rats was monitored continuously by laser-Doppler flowmetry (LDF) and reflectance spectrophotometry, and the validity of these techniques was determined. Corpus mucosal hemodynamics was recorded for 90 min, under stable conditions. In cases of graded hemorrhagic hypotension, corpus mucosal blood flow by LDF and hydrogen gas clearance, and potential differences showed a good correlation. Corpus, antral, and duodenal mucosal hemodynamics monitored by LDF, hydrogen gas clearance, and reflectance spectrophotometry reflected regional hemodynamic differences. In monitoring mucosal hemodynamics by LDF and reflectance spectrophotometry, regular oscillations (4-6 cycles/min) were observed in most animals. The characteristic change of oscillations during graded hemorrhagic hypotension was thus elucidated. In moderate hypotension (50-90 mmHg), high-amplitude and high-frequency (5-10 cycles/min) oscillations were observed, while in cases of severe hypotension (25-40 mmHg), the oscillations almost ceased. Observation of the oscillatory changes is thus a new application of LDF and reflectance spectrophotometry.     

Relationship between gastric mucosal hemodynamics and gastric motility

Continuous measurement of gastric mucosal hemodynamics (the index of mucosal hemoglobin concentration, the index of oxygen saturation and blood flow) in rats showed oscillatory changes. The mechanism of the oscillations was investigated using a probe specially designed for simultaneous measurement of hemodynamics and intragastric pressure. A hemodynamics-measuring probe for either reflectance spectrophotometry or laser-Doppler flowmetry was tied to a pressure microtransducer, inserted through an incision in the forestomach, and brought into gentle contact with the corpus mucosa. Synchronous oscillatory changes (4-6 cycles/min) in hemodynamics and motility were observed in the resting state (mean blood pressure: 120 mmHg). During moderate hemorrhagic hypotension (mean: 81 mmHg), oscillations in the hemodynamics increased in both amplitude and frequency, while motility remained constant. Oscillations in the hemodynamics were also affected by fluctuations in blood pressure and by topical application of norepinephrine to the corpus serosa. In water-immersion restraint rats, changes in the oscillations in the hemodynamics and motility were virtually synchronous; frequency decreased and amplitude increased. These findings suggest that oscillatory changes in gastric mucosal hemodynamics are regulated not only by gastric motility but also by arteriolar vasomotion of the gastric wall.     

Relationship between gastric mucosal hemodynamics and gastric motility

Continuous measurement of gastric mucosal hemodynamics (the index of mucosal hemoglobin concentration, the index of oxygen saturation and blood flow) in rats showed oscillatory changes. The mechanism of the oscillations was investigated using a probe specially designed for simultaneous measurement of hemodynamics and intragastric pressure. A hemodynamics-measuring probe for either reflectance spectrophotometry or laser-Doppler flowmetry was tied to a pressure microtransducer, inserted through an incision in the forestomach, and brought into gentle contact with the corpus mucosa. Synchronous oscillatory changes (4-6 cycles/min) in hemodynamics and motility were observed in the resting state (mean blood pressure: 120 mmHg). During moderate hemorrhagic hypotension (mean: 81 mmHg), oscillations in the hemodynamics increased in both amplitude and frequency, while motility remained constant. Oscillations in the hemodynamics were also affected by fluctuations in blood pressure and by topical application of norepinephrine to the corpus serosa. In water-immersion restraint rats, changes in the oscillations in the hemodynamics and motility were virtually synchronous; frequency decreased and amplitude increased. These findings suggest that oscillatory changes in gastric mucosal hemodynamics are regulated not only by gastric motility but also by arteriolar vasomotion of the gastric wall.     

Comparison of blood flow measurements by hydrogen gas clearance and laser Doppler flowmetry in the rat duodenum

This report examines the relationship between hydrogen gas clearance and laser Doppler flowmetry measurements in the duodenum of fasted, anesthetized rats under conditions of 1) reduced perfusion due to graded levels of hemorrhagic hypotension or 2) hyperemia due to perfusion with step doses of acid. There was a significant correlation between hydrogen gas clearance and laser Doppler flowmetry measurements (r = 0.73, p less than 0.01; n = 32 data points in 16 rats). The change in laser Doppler flowmetry values from the period immediately before to the period during the 3 min of acid perfusion was significantly correlated with the dose of acid used (r = 0.51, p less than 0.01; n = 27 rats). The changes in hydrogen gas clearance and laser Doppler flowmetry values from the 30-min period before to the 30-min period after acid perfusion were not correlated with the dose of acid used (r = 0.30 and 0.33, respectively). We conclude that in the rat duodenum 1) the significant linear correlation between hydrogen gas clearance and laser Doppler flowmetry when blood flow is reduced suggests that the countercurrent exchange mechanism is unlikely to modulate significantly hydrogen gas clearance measurements, and 2) the dose-related acid-induced duodenal hyperemia is transient rather than persistent when the rat duodenum is exposed to hydrochloric acid (0.03 to 0.1 N) for 3 min.     

Pancreatic proteases and intestinal mucosal injury after ischemia and reperfusion in the pig.

Intraluminal pancreatic proteases have been proposed to play a pathogenic role in the injury seen after ischemia and reperfusion of the small intestinal mucosa. Intestinal ischemia can be detected by indirect intramucosal pH measurements using tonometry. In this study, pigs were subjected to laparotomy and ligation of the pancreatic duct (n = 10) or a sham procedure (n = 10). Three weeks later, a standardized hemorrhagic shock was induced followed by retransfusion. Central hemodynamics, portal venous flow, and duodenal and small intestinal mucosal intramucosal pH were monitored. Samples were obtained from the small intestine for microscopic examination. A typical superficial mucosal injury developed in both groups of animals after reperfusion. However, the injury developed significantly later in the duct-ligated animals. No major differences in survival, splanchnic hemodynamics, or intramucosal pH between the groups were seen during hemorrhagic hypotension or after reperfusion. These data favor the concept that intraluminal pancreatic proteases are important for the rapid development of the mucosal reperfusion injury.     

Burn-Induced Gastric Mucosal Hemodynamic Disturbance in the Rat: Role of Platelet-Activating Factor

The role of platelet-activating factor (PAF) as a mediator of gastric damage associated with burn injury was examined in the rat. Gastric mucosal hemodynamics was recorded continuously by means of laser-Doppler flowmetry and reflectance spectrophotometry. Burn injury induced prolonged hypotension and rapid-onset and long-lasting gastric mucosal hemodynamic derangement—that is, ischemia with congestion. This hemodynamic disturbance was significantly inhibited by CV-6209, a specific PAF antagonist. Elevated tissue levels of thiobarbituric acid reactants and pathologic changes in the gastric mucosa subsequent to burn injury were also significantly alleviated by the preadministration of CV-6209. These results strongly suggest a role of PAF as an important mediator of gastric damage after burn injury through its potent vasoactive effect on gastric microcirculation.     

Burn-induced gastric mucosal hemodynamic disturbance in the rat. Role of platelet-activating factor.

The role of platelet-activating factor (PAF) as a mediator of gastric damage associated with burn injury was examined in the rat. Gastric mucosal hemodynamics was recorded continuously by means of laser-Doppler flowmetry and reflectance spectrophotometry. Burn injury induced prolonged hypotension and rapid-onset and long-lasting gastric mucosal hemodynamic derangement-that is, ischemia with congestion. This hemodynamic disturbance was significantly inhibited by CV-6209, a specific PAF antagonist. Elevated tissue levels of thiobarbituric acid reactants and pathologic changes in the gastric mucosa subsequent to burn injury were also significantly alleviated by the preadministration of CV-6209. These results strongly suggest a role of PAF as an important mediator of gastric damage after burn injury through its potent vasoactive effect on gastric microcirculation.     

Oxygen supply to the liver in patients with alcoholic liver disease assessed by organ-reflectance spectrophotometry

In the present study we have investigated hepatic hemodynamics in patients with alcoholic liver disease using reflectance spectrophotometry and the hydrogen clearance method. Analysis of 38 cases has shown that estimated regional hepatic-tissue hemoglobin concentration, expressed as a difference in absorbance between 569 and 650 nm (delta Er569-650), decreased significantly with progress of fibrosis or fat accumulation in the liver. This suggests that the relative compression of the vascular compartment is due to the progress of alcoholic liver disease. Estimated hepatic hemoglobin concentration also correlated positively with prothrombin time, and negatively with serum gamma-globulin level and 15-min retention rate of indocyanine green. The difference in absorbance between 569 and 650 nm obtained by reflectance spectrophotometry was positively correlated with the regional hepatic blood flow as measured by the hydrogen clearance method. Thus, it is concluded that the estimated regional hepatic-tissue hemoglobin concentration decreases with progress of fibrosis and fat accumulation in the liver, and that this decreased oxygen supply to the liver may have an important role in the progress of alcoholic liver disease.     

Interaction between hemorrhagic hypotension and hypoxia in regulation of renal vascular resistance in unanesthetized rabbits

Effects of interaction between hemorrhagic hypotension and hypoxia on the renal circulation were examined in awake rabbits. The hypothesis tested was that renal vasoconstriction during hemorrhagic hypotension is affected by arterial PO2 (PaO2). Awake rabbits were placed into an environmental chamber and exposed to either normoxia (PaO2 greater than 100 mmHg) or hypoxia (PaO2 less than or equal to 40 mmHg). Renal blood flow (RBF) was measured with 15 micron microspheres during normotension (mean arterial pressure = 86-97 mmHg), moderate hemorrhagic hypotension (mean arterial pressure = 62-65 mmHg), and severe hemorrhagic hypotension (mean arterial pressure = 49-50 mmHg). During normotension, RBF was 461 +/- 46 and 330 +/- 30 ml/min per 100 g (mean +/- SEM) in the normoxic and hypoxic groups, respectively (P less than 0.05), and renal vascular resistance (RVR) was 0.21 +/- 0.03 and 0.31 +/- 0.04 mmHg per ml/min per 100 g in the normoxic and hypoxic groups, respectively (NS). Renal blood flow fell progressively in each group during hypotension, but the magnitude of this response was unaffected by the level of PaO2. In contrast, RVR during severe hypotension was 0.42 +/- 0.11 and 2.08 +/- 0.57 mmHg per 100 g for normoxia and hypoxia, respectively (P less than 0.05), and the increase in RVR during severe hypotension compared to normotension was greater in the hypoxic than normoxic group (P less than 0.05). Thus, in unanesthetized rabbits, hypoxia reduces RBF but does not change RVR significantly during normotension, and hypoxia potentiates the increase in RVR during hemorrhagic hypotension.     

Hemodynamic and metabolic effects of ethanol in canine hemorrhagic shock

Ethanol has been reported to cause myocardial suppression, exaggerated hypotension, and increased mortality in various animal models of hemorrhagic shock. Previous studies have not used a fixed-volume graded hemorrhage model and have not monitored cardiac output or metabolic parameters such as serum glucose and lactate levels. We studied hemodynamic and metabolic changes after administration of ethanol in a 50% graded hemorrhage model in conditioned, anesthetized beagles after orogastric ethanol loading. The hemorrhage was done over a 60-min period followed by a 90-min stabilization period. The ethanol group (n = 6) had significantly higher heart rates during the stabilization period. Mean arterial pressures (MAP) were lower in the ethanol group during the stabilization period. The change from baseline MAP 30 min after hemorrhage was -31% in the control group and -53% in the ethanol group (P less than .05 using Wilcoxon ranked sum test). Serum glucose and lactate levels were higher in the ethanol group. These results indicate that ethanol impairs hemodynamics and alters glucose and lactate metabolism in dogs in the fixed-volume graded hemorrhage model. The effect of these changes on morbidity and mortality remains to be determined.     

Hemodynamic effects of naloxone on hemorrhagic shock in the beagle

Naloxone reverses the hypotension in various types of hemorrhagic shock models. What has yet to be firmly established is the mechanism by which naloxone reverses the hypotension. In a canine hemorrhagic shock model, impedance cardiography and invasive methods were used to measure various cardiovascular parameters. All dogs (beagles, 10-15 kg) were bled to and maintained at a mean arterial blood pressure (MAP) of 60 mmHg for 90 min and were then given either naloxone (2 mg/kg; n = 6) or an equivalent volume of saline (n = 6) intravenously (IV). After another 90 min observation period, the shed blood was reinfused. No significant differences in the preshock and shock cardiodynamics were noted between the naloxone and the control animals. During the treatment period, MAP was significantly increased in the naloxone group. There was no increase in cardiac output (CO), stroke volume (SV), end diastolic volume (EDV), dP/dt max, dP/dt/P, or HI (the impedance contractility index) over control animals. The most significant parameter improvement was total peripheral resistance (TPR). The data suggest that naloxone in this hemorrhagic shock model improves hemodynamics primarily by increasing vascular resistance.     

Regional gastric mucosal blood flow measurements by hydrogen gas clearance in the anesthetized rat and rabbit

Hydrogen gas clearance using 3% hydrogen in air and platinum contact electrodes was employed for measuring antral and corpus mucosal blood flow in anesthetized animals. Significantly greater antral than corpus mucosal blood flow was consistently demonstrated. Corpus but not antral mucosal blood flow showed a significant dose-related increase with intravenous pentagastrin. Vasopressin induced a significant dose-related decrease in both antral and corpus mucosal blood flow. Simultaneous measurement of basal corpus mucosal blood flow by hydrogen gas clearance and of gastric mucosal blood flow by aminopyrine clearance gave similar values, but the changes with intravenous pentagastrin or vasopressin measured by aminopyrine clearance were of a much higher order of magnitude. Hydrogen gas clearance, however, reflected changes in left gastric artery blood flow much more closely than did aminopyrine clearance. Therefore, we conclude that the hydrogen gas clearance technique as described is valid for measuring regional gastric mucosal blood flow. It is safe and has potential application in human studies.     

Rectal mucosal hemodynamics, evaluated by reflectance spectrophotometry, in patients with chronic hepatitis

To evaluate rectal mucosal hemodynamics in patients with chronic hepatitis, we employed reflectance spectrophotometry and examined the results in relation to the presence and severity of chronic hepatitis. Twenty-six patients with histologically diagnosed chronic hepatitis and 21 controls were examined for rectal vascular findings by endoscopy. Indices (I) of rectal mucosal oxygen saturation (ISO₂) and rectal mucosal hemoglobin (IHb) concentration were measured. To minimize the effects of systemic anemia, the IHb was divided by blood Hb concentration, giving the rectal index for Hb (RHb). The relationship between rectal mucosal hemodynamics and the histological grade of chronic hepatitis was studied. Rectal vascular lesions were observed in three patients with chronic hepatitis (11.5%). The RHb in patients with chronic hepatitis was significantly higher than that in the controls (5.74 ± 0.71 and 4.82 ± 1.12, respectively; P < 0.01). There was no significant difference in ISO₂ levels (44.23 ± 5.84 and 41.94 ± 4.91, respectively). No significant correlation was observed between rectal mucosal hemodynamics and the histological severity of chronic hepatitis, although rectal mucosal hemodynamics changed in patients with chronic hepatitis. Early vascular changes were observed in the rectal mucosa of patients with chronic hepatitis. (Received Oct. 23, 1997; accepted Dec. 19, 1997)     

Effects of experimental hypotension on hemodynamics and renin secretion rate

The effects of hypotension on systemic and renal hemodynamics, plasma renin activity (PRA), and renin secretion rate (RSR) were determined in dogs anesthetized with sodium pentobarbital plus chloralose. Renal blood flow (RBF) was determined with microspheres (15 micron) and with an electromagnetic flowmeter connected to an extra-corporeal circuit from the femoral artery to the renal artery. Hypotension was induced by nitroprusside infusion, which decreased peripheral resistance, and by hemorrhage, which reduced cardiac output. RSR increased in both forms of hypotension, but the increase following hemorrhage was greater than that after nitroprusside. Thus, when the mean arterial pressure (MAP) was reduced to 75 mmHg, RSR increased from 470 +/- 26 units/min to 990 +/- 12 units/min with nitroprusside and from 415 +/- 13 units/min to 1,509 +/- 21 units/min following hemorrhage. At MAP of 50 mmHg, RSR increased to 1,541 +/- 64 units/min with nitroprusside and to 2,254 +/- 98 units/min following hemorrhage. Nitroprusside increased renin secretion not only by an increase in sympathetic beta adrenergic activity through the baroreceptor reflex, but also by its direct vasolidatory effect in the renal circulation. In hemorrhagic hypotension, the increase in renin secretion was accompanied by renal vasoconstriction. The greater increase in RSR following hemorrhage than after nitroprusside at given levels of hypotension may be explained by a stronger beta adrenergic activation, the activation of prostaglandin and kallikrein systems, a lower microvascular pressure level, and/or smaller pulse pressure and lower sodium load in the macula densa. The comparison of renin secretion at the same degree of hypotension induced by different hemodynamic alterations serves to elucidate the mechanisms of renin secretion.     

The incidence and mechanism of hypotension following thrombolytic therapy for acute myocardixal infarction with streptokinase-containing agents -- lack of relationship to pretreatment streptokinase resistance

The incidence, amplitude, mechanism and relationship to priorexposure to streptococcal antigen of bloodpressure changes tostreptokinase-containing thrombolytic agents were investigatedin 125 patients treated with either 15 x 10⁶ IU streptokinaseover 60 min or 30 U anistreplase over 5 min, within 6 h of onsetof acute myocardial infarction. Twenty-one of 52 patients withanterior and 34 of 73 with inferior myocardial infarction hada hypotensive response. There were no signficant differencesin the incidence, duration or amplitude of hypotension betweenthe two treatment groups. The maximum mean fall in systolicblood pressure was 16.9 mmHg (95% confidence limits, CL 12.2to 24.5 mmHg), and the maximum mean fall in diastolic bloodpressure was 13.7 mmHg (CL 10.3 to 17.1 mmHg), starting 4 minafter start of therapy and resolving within 34 min. Blood pressurechanges were well tolerated. Hypotension was not related topretreatment streptokinase resistance titre, or anti-SK IgGconcentration, to changes in plasma fibrinogen, B-ß15–42 peptide, D-dimer—as indices of thrombin activationand fibrin (-ogen) breakdown — to plasma viscosity. The blood pressure changes following treatment with streptokinase-containingthrombolytic agents in acute myocardial infarction are frequentbut well tolerated. The mechanism of hypotension remains unclear,but is not related to prior exposure to streptococcal antigen.     

Impaired gastric mucosal energy metabolism in congestive gastropathy in cirrhotic patients

To clarify the characteristics of congestive gastropathy, we investigated gastric mucosal hemodynamics and energy metabolism in cirrhotic patients, using a reflectance spectrophotometry system and high performance liquid chromatography. The index of the gastric mucosal blood volume of cirrhotic patients with esophageal varices was significantly higher, and the index of gastric mucosal blood oxygenation significantly lower, than those in controls, thus indicating congestion and hypoxia in the gastric mucosa. Energy charge levels in the gastric mucosa of cirrhotic patients with esophageal varices were also significantly decreased. The energy charge level showed a strong linear correlation with the index of mucosal blood oxygenation in the antral (r=0.996,P<0.01) and body (r=0.994,P<0.01) mucosa of the stomach. These findings suggest that congestive gastropathy in a portal hypertensive state causes hypoxia in the gastric mucosa, leading to a mucosal energy deficit that may increase mucosal susceptibility to aggressive factors.     

Gastric hemodynamic disturbance induced by hemorrhagic shock in rats. Role of platelet-activating factor

The role of platelet-activating factor (PAF) in the induction of rat gastric mucosal damage by ischemia-reperfusion was examined with reference to hemodynamic characteristics. Gastric mucosal hemodynamics was measured continuously, using laser-Doppler flowmetry and reflectance spectrophotometry. Not only gastric mucosal damage but also characteristic hemodynamic change--that is, ischemia with congestion--was observed in the PAF infusion and reperfusion periods of the ischemia-reperfusion model. CV-6209, a specific PAF antagonist, significantly alleviated gastric mucosal damage and hemodynamic disturbance induced by ischemia-reperfusion. These results strongly suggest that PAF is a potent mediator of gastric mucosal damage during reperfusion in the ischemia-reperfusion model.     

Intracellular calcium level, lipid peroxidation, and development of gastric mucosal injury in rat hemorrhagic shock

Although acute gastric mucosal lesions (AGMLs) develop after the mucosal ischemia and reperfusion, precise intracellular mechanism for the development of AGMLs remains unclear. In the present paper, we investigated intracellular calcium level, lipid peroxidation, and gastric mucosal lesion in rat hemorrhagic shock model. We estimated intracellular calcium by measuring phosphorylase a activity in the gastric mucosa. The mucosal phosphorylase a increased during hemorrhagic shock, while it returned to be normal after the reinfusion of shed blood. Although mucosal lipid peroxide and area of AGMLs did not increase significantly during hemorrhagic shock, they increased significantly after the reinfusion. Diltiazem, a calcium antagonist, prohibited both of the increase in mucosal phosphorylase a activity during hemorrhagic shock and the increase in lipid peroxide content and area of AGMLs after the reinfusion of the shed blood. Organ reflectance spectrophotometry revealed that the gastric corpus suffered mucosal ischemia and hypoxia during the hemorrhagic shock. The are of AGMLs had a good correlation with a level of lipid peroxidation in the gastric mucosa. These results suggest 1) that intracellular free calcium in gastric mucosa increases during the mucosal ischemia; 2) that a calcium antagonist prohibits the increased intracellular free calcium during mucosal ischemia; and 3) that a calcium antagonist prohibits the increased mucosal lipid peroxide and the development of AGMLs after the mucosal reperfusion. Thus it is concluded that the increased intracellular free calcium during ischemia plays a key role in the mucosal tissue injury in the ischemia-reperfusion state.     

Endoscopic demonstration that vasopressin but not propranolol produces gastric mucosal ischemia in dogs with portal hypertension

Endoscopic reflectance spectrophotometry was used to compare the effect of vasopressin and propranolol on gastric mucosal hemodynamics in dogs with surgically induced esophageal varices and prehepatic portal hypertension. Reflectance spectrophotometry provides indices of mucosal hemoglobin concentration (IHB) and oxygen saturation (ISO2). Hyperemia (increased IHB, normal ISO2), ischemia without congestion (decreased IHB, decreased ISO2), and ischemia with congestion (increased IHB, decreased ISO2) are accompanied by characteristic patterns of IHB and ISO2. Under anesthesia, measurements were obtained on separate days from the gastric corpus mucosa of eight dogs before and 2 to 10 min after either 1 to 5 units of intravenous vasopressin or 1 mg of propranolol. Results revealed that vasopressin (in doses that significantly reduced variceal and portal venous pressure in this animal model) produced a reduction in both IHB and ISO2, indicating gastric mucosal ischemia secondary to splanchnic vasoconstriction. On the other hand, propranolol in a dose that significantly reduced pulse rate by 27 +/- 2% had no effect on IHB or ISO2, suggesting that this dose of propranolol has no direct vasoactive effect on the gastric (splanchnic) circulation.     

Nicotine and smoking do not decrease basal gastric mucosal blood flow in anesthetized rats

The literature regarding the effect of nicotine and cigarette smoke on gastric blood flow is conflicting. The hydrogen gas clearance technique was used to measure the effects of nicotine and cigarette smoke on basal gastric mucosal blood flow in anesthetized rats. Blood flow was measured before, during, and after treatment with either intravenous nicotine (4 or 40 g/kg/min) or inhaled cigarette smoke (nicotine or nicotine free). Neither intravenous nicotine nor cigarette smoke significantly altered gastric mucosal blood flow. On the other hand, hypotension produced by hemorrhage significantly decreased mucosal blood flow (P<0.05). Thus the technique used could detect a decrease in blood flow. These findings indicate that in the anesthetized rats, hypotension but not intravenous nicotine or cigarette smoke, in the doses given, reduce gastric mucosal blood flow.This work was supported in part by NIAMDD grant AM 25891, and Veterans Administration Research Funds.