Increased Lung Microvascular Permeability Research Articles

Serum amyloid A3 confers protection against acute lung injury in Pseudomonas aeruginosa-infected mice.

Pseudomonas aeruginosa is a gram-negative bacterium associated with serious illnesses, including ventilator-associated pneumonia and various sepsis syndromes in humans. Understanding the host immune mechanisms against P. aeruginosa is, therefore, of clinical importance. The present study identified serum amyloid A3 (SAA3) as being highly inducible in mouse bronchial epithelium following P. aeruginosa infection. Genetic deletion of Saa3 rendered mice more susceptible to P. aeruginosa infection with decreased neutrophil superoxide anion production, and ex vivo treatment of mouse neutrophils with recombinant SAA3 restored the ability of neutrophils to produce superoxide anions. The SAA3-deficient mice showed exacerbated inflammatory responses, which was characterized by pronounced neutrophil infiltration, elevated expression of TNF-α, KC/CXCL1, and MIP-2/CXCL2 in bronchoalveolar lavage fluid (BALF), and increased lung microvascular permeability compared with their wild-type littermates. BALF neutrophils from Saa3 knockout mice exhibited reduced superoxide anion production compared with neutrophils from wild-type mice. Adoptive transfer of SAA3-treated neutrophils to Saa3 knockout mice ameliorated P. aeruginosa-induced acute lung injury. These findings demonstrate that SAA3 not only serves as a biomarker for infection and inflammation, but also plays a protective role against P. aeruginosa infection-induced lung injury in part through augmentation of neutrophil bactericidal functions.

BLOCKADE OF ENDOTHELIAL GROWTH FACTOR, ANGIOPOIETIN-2, REDUCES INDICES OF ARDS AND MORTALITY IN MICE RESULTING FROM THE DUAL-INSULTS OF HEMORRHAGIC SHOCK AND SEPSIS.

We have demonstrated hemorrhagic shock "priming" for the development of indirect acute respiratory distress syndrome (iARDS) in mice following subsequent septic challenge, and show pathology characteristic of patients with iARDS, including increased lung microvascular permeability and arterial PO2/FI02 reduced to levels comparable to mild/moderate ARDS during the 48 h following hemorrhage. Loss of endothelial cell (EC) barrier function is a major component in the development of iARDS. EC growth factors, Angiopoietin (Ang)-1 and 2, maintain vascular homeostasis via tightly regulated competitive interaction with tyrosine kinase receptor, Tie2, expressed on ECs. Ang-2/Tie2 binding, in contrast to Ang-1, is believed to produce vessel destabilization, pulmonary leakage, and inflammation. Recent clinical findings from our trauma/surgical intensive care units and others have reported elevated Ang-2 in the plasma from patients that develop ARDS. We have previously described similarly elevated Ang-2 in plasma and lung tissue in our shock/sepsis model for the development of iARDS, and demonstrated effective reduction in indices of inflammation and lung tissue injury following siRNA inhibition of Ang-2 protein synthesis. In this study we show that Ang-2 in lung tissue and plasma spikes following hemorrhage (priming) and remain elevated at sepsis induction. In addition, that transient inhibition of Ang-2 function immediately following hemorrhage, suppressing priming, but not following sepsis, impacts the development of iARDS in our model. Our data demonstrate that selective temporal blockade of Ang-2 function following hemorrhagic shock priming significantly improved PO2/FIO2, decreased lung protein leak and indices of inflammation, and improved 10-day survival in our murine model for the development iARDS.

Acute Effects of Estradiol on Lung Inflammation Due to Intestinal Ischemic Insult in Male Rats

Intestinal ischemia and reperfusion (intestinal I/R) causes acute lung inflammation that is characterized by leukocyte migration, increased lung microvascular permeability, and, in severe forms, noncardiogenic pulmonary edema and acute respiratory distress syndrome. Female sex hormones interfere with immune response, and experimental and clinical evidence shows that females are more resistant than males to organ injury caused by gut trauma. To reduce the lung inflammation caused by intestinal I/R, we have acutely treated male rats with estradiol. Intestinal I/R was performed by the clamping (45 min) of the superior mesenteric artery (SMA), followed by 2 h of intestinal reperfusion (unclamping SMA). Groups of rats received 17β estradiol (E2, 280 µg/kg, i.v., single dose) 30 min after the SMA occlusion (ischemia period) or 1 h after the unclamping of SMA (reperfusion period). Leukocytes influx into the lung and microvascular leakage were assessed by lung myeloperoxidase activity and Evans blue dye extravasation, respectively. The lung expression of adhesion molecules (intercellular adhesion molecule 1, platelet endothelial cell adhesion molecule 1, and vascular cell adhesion molecule [VCAM]) was evaluated by immunohistochemistry. Interleukin 1β (IL-1β), IL-10, and NOx concentrations were quantified in supernatants of cultured lung tissue. We have found that intestinal I/R increased the lung myeloperoxidase activity and Evans blue dye extravasation, which were reduced by treatment of rats with E2. Intestinal I/R increased ICAM-1 expression only, and it was decreased by E2 treatment. However, E2 treatment reduced the basal expression of platelet endothelial cell adhesion molecule 1. E2 treatment during intestinal ischemia was effective to reduce the levels of IL-10 and IL-1β in explant supernatant, but only IL-10 levels were reduced by E2 at reperfusion phase. The treatment with E2 did not affect NOx concentration. Taken together, our data suggest that estradiol modulates the lung inflammatory response induced by lung injury, likely by acute effects. Thus, acute estradiol treatment could be considered as a potential therapeutic agent in ischemic events.

Pulmonary Endothelial Cell Activation During Experimental Acute Kidney Injury

Acute kidney injury (AKI) leads to increased lung microvascular permeability, leukocyte infiltration, and upregulation of soluble inflammatory proteins in rodents. Most work investigating connections between AKI and pulmonary dysfunction, however, has focused on characterizing whole lung tissue changes associated with AKI. Studies at the cellular level are essential to understanding the molecular basis of lung changes during AKI. Given that the pulmonary microvascular barrier is functionally abnormal during AKI, we hypothesized that AKI induces a specific proinflammatory and proapoptotic lung endothelial cell (EC) response. Four and 24 h after kidney ischemia/reperfusion injury or bilateral nephrectomy, murine pulmonary ECs were isolated via tissue digestion followed by magnetic bead sorting. Purified lung ECs were analyzed for changes in mRNA expression using real-time SuperArray polymerase chain reaction analysis of genes related to EC function. In parallel experiments, confluent rat pulmonary microvascular ECs were treated with AKI or control serum to evaluate functional cellular alterations. Immunocytochemistry and FACS analysis of Annexin V/propidium iodide staining were used to evaluate cytoskeletal changes and promotion of apoptosis. Isolated murine pulmonary ECs exhibited significant changes in the expression of gene products related to inflammation, vascular reactivity, and programmed cell death. Further experiments using an in vitro rat pulmonary microvascular EC system revealed that AKI serum induced functional cellular changes related to apoptosis, including structural actin alterations and phosphatidylserine translocation. Analysis and segregation of both upregulated and downregulated genes into functional roles suggest that these transcriptional events likely participate in the transition to an activated proinflammatory and proapoptotic EC phenotype during AKI. Further mechanistic analysis of EC-specific events in the lung during AKI might reveal potential novel therapeutic targets for the deleterious kidney-lung crosstalk in the critically ill patient.

TRPC1‐Mediated Ca2+ Entry Increases Lung Microvascular Permeability

Despite the evidence that transient receptor potential canonical (TRPC1) channel can mediate Ca2+ entry in endothelial cells the role of TRPC1 activity in the mechanism of increased lung vascular permeability remains unclear. We isolated lungs from TRPC1−/− and wild type (WT) mice and activated Ca2+ entry by two independent mechanisms to determine the role of TRPC1 in regulating lung microvascular permeability. We perfused lungs with either thapsigargin, that activates Ca2+ entry by depleting intracellular stores or diacylglycerol (DAG), that activate Ca2+ entry independent of store depletion. We determined lung microvascular permeability by determining filtration co‐efficient (Kfc), a measure of hydraulic conductivity across endothelial exchange barrier. We show that in WT lungs thapsigargin increased lung Kfc whereas thapsigargin failed to increase Kfc in lungs isolated from TRPC1−/− mice. Interestingly, OAG, a cell permeable analog of DAG, increased Kfc in TRPC1−/− mice but the Kfc response was significantly lower than that observed in WT mice lungs. We next determined the contribution of TRPC1 in regulating lung microvascular permeability in response to thrombin ligation of protease activating receptor 1 (PAR‐1) which activate Ca2+ entry both by store‐dependent and DAG‐dependent manner. We show that PAR‐1 activation by specific PAR‐1 peptide increased Kfc in WT lungs whereas it failed to induce a response in lungs isolated from TRPC1−/− mice. We also address the role of TRPC1 in the lung inflammatory response to sepsis induced by i.p. injection of LPS. Prevention of lung injury in TRPC1−/− mice was associated with decreased mortality in response to LPS challenge. Thus, TRPC1 through its ability to regulate Ca2+ entry is a crucial determinant of increased lung microvascular permeability and inflammatory response.

Lung neutrophil retention and injury after intestinal ischemia/reperfusion.

To define the mechanisms responsible for the lung leukosequestration and injury elicited by intestinal ischemia/reperfusion (I/R). The effect of 120 minutes of superior mesenteric artery occlusion and 90 minutes of reperfusion on neutrophil deformability, lung neutrophil retention, and pulmonary microvascular permeability was determined. Compared with control surgery, intestinal I/R resulted in a significant increase in neutrophil stiffness (mean yield pressure [Pyield], 1.533 +/- 0.075 and 2.302 +/- 0.288 cm H2O, respectively) and lung neutrophil content (6.3 +/- 1.4 and 31.5 +/- 6.4 U/g wet weight, respectively). These changes were not affected by inhibition of neutrophil adherence before gut reperfusion. However, the increased lung microvascular permeability elicited by gut I/R (0.111 +/- 0.020 [control surgery] and 0.255 +/- 0.041 [I/R] mL/min/cm H2O/100 g lung tissue) was significantly attenuated by administration of antibodies directed against neutrophil or endothelial determinants of leukocyte adhesion. The results of this study suggest that intestinal I/R is a potent inflammatory stimulus that elicits an increase in neutrophil stiffness and lung neutrophil retention independent of neutrophil-endothelial cell adhesion. In contrast, the increased lung microvascular permeability elicited by gut I/R is attenuated by strategies that interfere with neutrophil-endothelial cell adhesion.

Ischemia and reperfusion injury: prevention of pulmonary hypertension and leukosequestration following lower limb ischemia

Ischemia is a common clinical event with potentially serious consequences. The major part of tissue damage occurs upon reperfusion and is mediated by activated neutrophils. Ischemia reperfusion injury is manifested by oedema and increased microvascular permeability. This study tested cardiopulmonary functions following 2 h of lower limb ischemia. Anesthetized dogs were randomized into three groups: nonischemic sham dogs ( n = 4), ischemic control dogs ( n = 8) and ischemic dogs pretreated with prostaglandin (PG)E 1 ( n = 8). In control animals, mean pulmonary artery pressure (mPAP) increased 1 min after declamping from 13.37 ± 2.61 mmHg to 16.88 ± 3.68 mm Hg ( P < 0.05). Pulmonary microvascular pressure (Pmv) increased within 1 minute of reperfusion from 7.71 ± 1.87 mm Hg to 10.54 ± 3.71 mm Hg ( P < 0.05). These changes are consistent with increased lung microvascular permeability. White blood cell count fell slightly but not significantly and lung histology showed leukosequestration in alveoli of 171 ± 22 polymorphonuclear leukocyte (PMN) 10 high powered fields (HPF) in the ischemic control group compared with 121 ± 56 PMN 10 HPF in the sham group ( P < 0.05). Systemic arterial pressure, cardiac output, central venous pressure and pulmonary artery wedge pressure were unaffected. In animals pretreated with PGE 1, mPAP and Pmv were unchanged during reperfusion. Lung histologic findings appeared normal and leukosequestration was not observed. PMN counts in alveoli showed 95 ± 26 PMN 10 HPF , lower than in ischemic control animals ( P < 0.05). These data indicate that lower limb ischemia during reperfusion leads to pulmonary hypertension and leukosequestration. PGE 1 infusion is effective in limiting ischemia reperfusion injury.

Pulmonary embolism as a cause of pulmonary oedema

Acute pulmonary oedema is a well-described complication of pulmonary embolism. However, the relationship between these two conditions is not widely appreciated by physicians and the diagnosis of an underlying pulmonary embolism in patients with pulmonary oedema may well be missed. We describe two cases in which pulmonary oedema complicated pulmonary embolism, and discuss the possible factors involved in the development of oedema. These include altered left ventricular (LV) function, increased lung microvascular permeability, overperfusion and reper-fusion injury. We also emphasise the importance of considering the possibility of an underlying pulmonary embolism in patients with pulmonary oedema so that appropriate thrombolytic therapy can be given early.

Acute lung injury following reperfusion after ischemia in the hind limbs of rats.

In this study, we proposed that oxygen free radicals participate in the acute pulmonary injury that follows limb ischemia/reperfusion. Using an established model of hind limb ischemia, reproducible lung injury occurred after reperfusion. Lung microvascular permeability was measured with 125I-BSA and increased two-fold after 30 minutes of reperfusion. Pulmonary injury was blocked with DMSO, DMTU, allopurinol, indomethacin, and SOD plus catalase. The degree of pulmonary neutrophil sequestration as assessed by tissue myeloperoxidase activity was significantly diminished in animals pretreated with antioxidants. Pretreatment with indomethacin did not attenuate the neutrophil sequestration within the pulmonary parenchyma. These data suggest that increased lung microvascular permeability and neutrophil accumulation occur following hind limb ischemia/reperfusion. Therapeutic interventions with oxygen radical inhibitors blocked this process, while the prostaglandin inhibitor, indomethacin, only reduced lung permeability.

Thromboxane A2 mediates increased pulmonary microvascular permeability following limb ischemia.

Lower torso ischemia and reperfusion lead to respiratory dysfunction characterized by pulmonary hypertension and increased lung microvascular permeability. This is associated with lung leukosequestration and thromboxane (TX) generation. This study tests the role of elevated TX levels following muscle ischemia in mediating remote lung injury. Anesthetized sheep prepared with chronic lung lymph fistulae underwent 2 hours of bilateral hind limb tourniquet ischemia. In untreated controls (n = 7), 1 minute after reperfusion there was a transient increase in plasma immunoreactive (i)-TXB2 levels from 211 to 735 pg/ml (p less than 0.05), and at 30 minutes, lung lymph i-TXB2 levels rose from 400 to 1,005 pg/ml (p less than 0.05). At 1 minute, the mean pulmonary arterial pressure (MPAP) increased from 13 to 38 mm Hg (p less than 0.05) and pulmonary microvascular pressure (Pmv) from 7 to 18 mm Hg (p less than 0.05). Lung lymph flow (QL) rose from 4.3 to 8.3 ml/30 min (p less than 0.05), the lymph/plasma (L/P) protein ratio was unchanged from 0.6, and the lymph protein clearance increased from 2.6 to 4.6 ml/30 min (p less than 0.05). Two hours after reperfusion, neutrophils were observed sequestered in lung capillaries and proteinaceous exudates were found in alveoli in contrast to sham-operated animals (n = 3). To maximize lung vascular surface area and achieve a pressure independent L/P protein ratio a left atrial balloon was inflated during one group of ischemia-reperfusion experiments (n = 5). This resulted in a baseline rise in MPAP to 20 mm Hg (p less than 0.05); a 4.3-fold increase in QL (p less than 0.05), a decrease in the L/P ratio from 0.70 to 0.28 (p less than 0.05) and a protein reflection coefficient (sigma d) of 0.72. During reperfusion the L/P ratio rose to 0.49 (p less than 0.05) and the sigma d decreased to 0.51 (p less than 0.05), documenting an increase in lung microvascular permeability. In contrast to untreated ischemic controls, inhibition of TX synthetase with OKY 046 (n = 6) reduced plasma i-TXB2 levels to 85 pg/ml (p less than 0.05) but also increased i-6-keto-PGF1 alpha levels to 78 pg/ml relative to 15 pg/ml in untreated controls (p less than 0.05). OKY 046 prevented the increase in MPAP, Pmv, QL, and lymph protein clearance (p less than 0.05). Lung histology was normal in distinction to the leukosequestration in untreated ischemic controls.(ABSTRACT TRUNCATED AT 400 WORDS)

Verapamil attenuates lung vascular responses to endotoxin in sheep

Experiments were conducted on five chronically instrumented unanesthetized sheep to determine the effects of verapamil, a calcium channel inhibitor, on the pulmonary hemodynamic and microvascular permeability responses to endotoxemia. Paired control endotoxemia experiments (E) and endotoxemia with verapamil treatment (30-60 micrograms.kg-1.min-1) experiments (V + E) were conducted on each sheep in random order. In the V + E experiments sheep were pretreated with a continuous intravenous infusion of verapamil 1.5-2.0 h before endotoxin infusion (1.0 microgram/kg, given over 15 min). Verapamil significantly increased base-line pulmonary arterial pressure, left atrial pressure, lung lymph flow rate, and circulating blood leukocyte levels and significantly decreased base-line cardiac output. During the endotoxin response, verapamil significantly attenuated both phase I pulmonary arterial hypertension and phase II lung lymph flow rate compared with control endotoxin experiments. The results indicate that verapamil attenuates both the pulmonary hemodynamic and increased lung microvascular permeability response to endotoxin in sheep. In a series of in vitro experiments, verapamil was found to be a potent inhibitor of phorbol myristate acetate-induced superoxide production in isolated sheep granulocytes. These data suggest that the beneficial in vivo effects of verapamil during endotoxemia may in part be due to its inhibition of increased free cytosol calcium concentration and/or inhibition of toxic O2 metabolite production.

Inhalation injury in burned patients: effects and treatment

Pulmonary pathology in major thermal injury is found in 30–80 per cent of burn fatalities. The incidence and mortality from inhalation injury increases both with age and increasing burn size. Toxic smoke inhalation injury, characterized by increased lung microvascular permeability, is attenuated by increasing cardiac output to normal levels, indicating that fluid restriction after inhalation injury may lead to excessive lung fluid formation and hypoxia. Fluid administration of approximately 2ml/kg/% area burned above the calculated resuscitation volume is required following an inhalation injury to provide adequate support for the systemic circulation and maintain cardiac output at normal levels. This additional volume does not contribute to the development of pulmonary oedema, but may decrease its formation by increasing shear forces thus reducing polymorphonuclear leucocyte deposition in the pulmonary microcirculation. Nasotracheal intubation is preferred when airway integrity is compromised by inhalation injury. The advantages are non-operative placement, ease of discontinuation, minimal bacterial contamination and leaving neck burns undisturbed. The administration of the appropriate antibiotics for documented infection is recommended, while steroids have been shown to be of no benefit.

Local pulmonary activation of proteolytic enzymes after Escherichia-coli-induced lung injury in sheep.

Activation of several cascade systems, e.g. coagulation, fibrinolysis, kallikreinkinin, complement and eicosanoid systems, has been implicated in the etiology of septic-lung microvascular injury. A chronic lung lymph fistula preparation in sheep (n = 9) was used to study coagulation, kallikrein-kinin and eicosanoids during Escherichia coli septicemia. Lung lymph flow and lymph composition indicated an increased lung microvascular permeability approximately 2 h after infusion of bacteria. Stable prothrombin and antithrombin III levels in lymph contradicted local activation of the coagulation cascade in the lung and systemic activation was not evident until 4 h after bacteria infusion. Lymph thromboxane B2 and 6-keto PGF1 alpha peaked early (1 h). Reduced lymph prekallikrein and kallikrein inhibitors indicated local activation of this system in the lung. Systemic activation of kallikrein could not be demonstrated. Thus, (1) changes in systemic blood may not adequately reflect local events and (2) studies of proteolytic enzymes and other inflammatory mediators in lung may contribute to clarifying the etiology of microvascular injury.

Perilla ketone: a model of increased pulmonary microvascular permeability pulmonary edema in sheep.

A model of increased microvascular permeability pulmonary edema was developed in chronically instrumented unanesthetized sheep using perilla ketone (PK). PK did not cause changes in pulmonary hemodynamics but did cause marked increases in the flow of protein-rich lung lymph. The changes in lung lymph flow were accompanied by radiographic evidence of both interstitial and alveolar pulmonary edema as well as hypoxemia. PK did not cause acute changes in lung mechanics. Dynamic compliance of the lungs and FRC decreased later, concomitant with the changes in lung lymph flow, radiographic evidence for pulmonary edema, and hypoxemia. Resistance to air flow across the lungs and specific conductance did not change significantly after PK infusion. The severity of the radiographic evidence for pulmonary edema observed after PK correlated with the severity of the concomitant hypoxemia and changes in dynamic compliance of the lungs. PK did not cause increases in the concentrations of cyclooxygenase products of arachidonic acid in lung lymph or plasma or changes in blood leukocyte counts. We conclude that PK causes increased lung microvascular permeability pulmonary edema without acute changes in pulmonary hemodynamics. This model permits study of the pathophysiologic aspects of increased lung microvascular permeability without the concomitant functional alterations that complicate most other experimental models of diffuse lung injury.

Prostaglandin E1 prevents increased lung microvascular permeability during intravascular complement activation in sheep.

Prostaglandin E1 (PGE1) inhibits a variety of functions of activated neutrophils including respiratory burst, release of leukotriene B4, and adherence to endothelial cells. To determine if PGE, alters the pathophysiology of complement-induced lung vascular injury, experiments were conducted in anesthetized sheep with lung lymph fistulas given a 1-hour infusion of zymosan-activated plasma. PGE1 (30 ng/min/kg) or its saline vehicle was infused intravenously for 90 minutes beginning 30 minutes before the infusion of activated plasma. PGE1 had no effect on leukocyte count, the initial hypoxemia and thromboxane A2 release, or the development of acute pulmonary hypertension. However, PGE1 prevented steady-state increases in lung lymph flow that in vehicle-treated sheep signaled an increase in lung microvascular permeability. Furthermore, extraction of PGE1 by pulmonary endothelial cells was unaffected by the infusion of activated plasma. We propose that PGE1 prevented the increase in lung vascular permeability by inhibiting adherence of activated neutrophils to endothelial cells.

Computer analysis of proteins separated by polyacrylamide gradient pore gel electrophoresis

A method for measuring lymph-to-plasma (L/P) protein concentration ratios obtained from protein fractions separated by polyacrylamide gradient gel electrophoresis is presented. A curve-fitting technique is used to decompose lymph and plasma electropherograms containing multiple components into individual components, eliminating protein-protein overlap regions. This allows the concentration of each component in the mixture to be measured accurately, yielding more precise estimates of L/P ratios. This technique consists of three phases. (1) Individual electropherograms are constructed for proteins of various sizes by taking a weighted average of measured electropherograms obtained from the two protein standards closest in size to the protein of interest, (2) Using these generated standard curves, the multicomponent lymph and plasma curves are decomposed into the least number of equally spaced components that yield a good fit. A linear least-squares method is used to do this. Each protein fraction is multiplied by the total measured protein concentration to provide a concentration for each component. (3) Finally, L/P concentration ratios of protein fractions with visible peaks were computed by applying an averaging technique to the equally spaced protein fractions. Plots of sheep lung L/P ratio versus protein size obtained in this manner were compared to L/P ratios obtained using a method of analysis which does not correct for protein overlap. The corrected L/P ratios showed less scatter than the uncorrected curves. Lung lymph data analyzed with the correction method indicated an increased lung microvascular permeability for large proteins following endotoxin infusion, whereas the uncorrected curves were too noisy to support this concept.

Changes in lung vascular permeability after heart-lung transplantation

We have employed multiple indicator dilution techniques (MID) in six patients after heart-lung transplantation to assess changes in the lung vascular permeability-surface area product for urea (PS). Serial PS values for the patients when normalized to the predicted total lung capacity (TLC) in liters, ranged between 1.04 and 6.27 ml/sec/TLC (patient 1), 0 and 2.76 ml/sec/TLC (patient 2), 0.59 and 2.88 ml/sec/TLC (patient 3), 0.13 and 1.23 ml/sec/TLC (patient 4). The elevated values for PS in patient 1 exceed the lethal range described by K. L. Brigham et al. ( J. Clin. Invest. 72: 339, 1983) for severe ARDS. This strongly suggests a severely increased lung microvascular permeability in this patient possibly secondary to rejection as indicated by endomyocardial biopsy. PS values for surviving patients 2–6 fell well below the corresponding lethal value for ARDS patients. We conclude that PS urea derived from MID provides an indicator of the status of lung microvascular integrity in heart-lung transplant recipients.

Superoxide dismutase with heparin prevents increased lung vascular permeability during air emboli in sheep.

We studied the effects of bovine superoxide dismutase on the increased lung microvascular permeability to fluid and protein during air emboli in unanesthetized sheep. We measured pulmonary arterial and left atrial pressures, cardiac output, lung lymph flow, and lymph and plasma protein concentrations. In air emboli experiments we continuously infused the same dose of air bubbles 1 mm in diameter into the right atrium of each sheep to increase pulmonary vascular resistance to 2-4 times base-line values. We did experiments in pairs so that each sheep served as its own control. We found an increase in protein-rich lymph flow from the lung during embolization in untreated sheep, indicating an increase in microvascular permeability to both fluid and protein. When we pretreated the sheep with heparin (300 U/kg) and infused them with intravenous bovine superoxide dismutase (1 mg X kg-1 X h-1 beginning before the air infusion), we found that vascular pressures and pulmonary vascular resistance were not different from untreated sheep but that the expected increases in transvascular fluid and protein flow during emboli were significantly decreased (P less than 0.0005). Heparin alone did not significantly attenuate the increased microvascular permeability but we found that it greatly enhanced the effectiveness of superoxide dismutase in preserving microvascular functional integrity during air emboli. We conclude that superoxide anion, probably produced and released by leukocytes, is a central factor in the microvascular injury that results in increased permeability in the lungs of sheep during air microembolization.

Dog lymph flow in increased capillary permeability states

The lung lymph flow rate ( Q ̇ L ) is increased in edema caused by an increase in lung microvascular permeability. This increase in Q ̇ L could be caused by either a decrease in the effective resistance of the lymph vessels ( R L), or by an increase in the effective lymph driving pressure ( P L), or by a change in both R L and P L. We estimated P L and R L from the linear relationship between Q ̇ L and the pressure at the outflow end ( P O) of five cannulated dog lung lymph vessels ( R L = −ΔP O Δ Q ̇ L and P L = the P O at which Q ̇ L = 0 ). We increased lung microvascular permeability by giving the dogs 100 mg/kg of alloxan and found that Q ̇ L increased from 24.5 ± 8.9 μl/min to 112 ± 41 μl/min (mean ± SD). R L decreased from 0.35 ± 0.12 to 0.11 ± 0.04 cm H 2O min/μl and P L increased from 8.5 ± 1.5 to 15.9 ± 2.7 cm H 2O. We then increased the capillary pressures from 18.3 ± 3.8 to 41.3 ± 7.3 cm H 2O and Q ̇ L increased to 169.9 ± 47.8 μl/min. P L increased by an additional 6.3 cm H 2O but R L decreased by only an additional 0.02 cm H 2O min/μl. These results show that the Q ̇ L vs P O relationship is changed in edema secondary to an increase in microvascular permeability, and that this change can be represented as changes in R L and P L. In terms of these parameters, Q ̇ L increased in edema as a result of a decrease in R L and an increase in P L.

Leukocytes are required for increased lung microvascular permeability after microembolization in sheep.

We studied the effects of uneven pulmonary artery obstruction by microemboli on steady state transvascular fluid and protein exchange in normal and leukopenic sheep. We measured pulmonary artery and left atrial pressures, cardiac output, lung lymph flow, and lymph plasma protein concentrations. Sheep were made profoundly leukopenic by administration of intra-arterial mechlorethamine hydrochloride (0.4 mg/kg, two doses) and colchicine (0.1-0.2 mg/kg, anesthetized sheep only). In anesthetized sheep, we injected glass beads 200 micrometers in diameter via the right atrium to raise pulmonary vascular resistance to 2-3 times baseline values. With normal levels of circulating leukocytes, sheep developed an increased protein-rich lymph flow from the lung characteristic of increased permeability edema. Leukopenic sheep had a significantly attenuated response after embolization for equivalent degrees of vascular obstruction. In unanesthetized sheep, we continuously infused air bubbles 1 mm in diameter via the right atrium to raise pulmonary vascular resistance to about 2 times baseline values. Each sheep served as its own control. With normal circulating leukocyte levels, there was an increase in protein-rich lymph flow from the lung during embolization. When the air infusion ended, the sheep recovered to the baseline condition in 24 hours. We induced emboli with the same amount of air when the sheep were profoundly leukopenic; lymph and protein flow from the lung were significantly less for equivalent degrees of obstruction. We conclude that circulating leukocytes are essential for the microvascular injury that results in increased permeability in the lungs of sheep after microembolization.