Alveolar and lung liquid clearance in anesthetized rabbits

Alveolar and lung liquid clearance were studied over 8 h in intact anesthetized ventilated rabbits by instillation of either isosmolar Ringer lactate (2 ml/kg) or autologous plasma (2 or 3 ml/kg) into one lower lobe. The half time for lung liquid clearance of the isosmolar Ringer lactate was 3.3 h and that for plasma clearance was 6 h. In the plasma experiments, the alveolar protein concentration after 1 h was 5.2 +/- 0.8 g/dl, which was significantly greater than the initial instilled protein concentration of 4.3 +/- 0.7 g/dl (P less than 0.05). Thus alveolar protein concentration increased by 21 +/- 12% over 1 h, which matched clearance from the entire lung of 19 +/- 11% of the instilled volume. Overall the rate of alveolar and lung liquid clearance in rabbits was significantly faster than in prior studies in dogs and sheep. The fast alveolar liquid clearance rate in rabbits was not due to higher endogenous catecholamine release, because intravenous and alveolar (5 x 10(-5) M) propranolol did not slow the clearance. Also, beta-adrenergic therapy with alveolar terbutaline (10(-5) or 10(-4) M) did not increase the alveolar or lung liquid clearance rates. Phloridzin (10(-3) M) did not slow alveolar liquid clearance. However, amiloride (10(-4) M) inhibited 75% of the basal alveolar liquid clearance in rabbits, thus providing evidence that alveolar liquid clearance in rabbits depends primarily on sodium-dependent transport. This rabbit study provides further evidence for important species differences in the basal rates of alveolar liquid and solute clearance as well as the response to beta-adrenergic agonists and ion transport inhibitors.     

Alveolar pressure inhomogeneity during low-frequency oscillation of excised canine lobes

To quantify the inhomogeneity of alveolar pressures (PA) during cyclic changes in lung volume similar to those present during spontaneous breathing, inhomogeneity of PA was measured with an alveolar capsule technique in six excised canine lungs. The lungs were ventilated by a quasi-sinusoidal pump with a constant end-expiratory lung volume and tidal volumes of 10, 20, and 40% of vital capacity at breathing frequencies ranging from 5 to 45 breaths/min. Inhomogeneity of PA was quantified as the sample standard deviation of pressures measured in three capsules. A component of inhomogeneity in phase with flow and a smaller component out of phase with flow were present. The in-phase component increased approximately linearly with flow. The ratio of inhomogeneity to flow was smaller at large tidal volumes and, at the two higher tidal volumes studied, the ratio was greater during inspiration than during expiration. If these data are interpreted in terms of a simple circuit model, this degree of inhomogeneity implies an approximately twofold variation in regional time constants. Despite these considerable differences in time constants, the absolute amount of inhomogeneity as defined by the sample standard deviation of the three PA's was small (maximum 0.57 +/- 0.32 cmH2O at the highest breathing frequency and tidal volume) because airway resistance in the canine lung was small.     

Alveolar liquid and protein clearance from normal dog lungs

To determine whether liquid and protein clearance from the air spaces and lungs of anesthetized and unanesthetized dogs is the same as in sheep, we quantified these variables at three different time periods (4, 8, and 12 h) by instilling heparinized plasma (3 ml/kg) labeled with 125I-albumin into one lower lobe. Protein clearance, measured from the residual 125I-albumin in the lung homogenate, was slow and monoexponential (approximately 1%/h), similar to our previous data for protein clearance from the lungs in sheep. Lung liquid clearance in dogs, however, was 50% less than in previous experiments in sheep. Residual lung liquid (as percent of instilled) was 88.7 +/- 7.0 at 4 h, 70.5 +/- 9.1 at 8 h, and 64.0 +/- 5.8 at 12 h. At each time period, alveolar protein concentration increased by 0.6 +/- 0.4 g/dl at 4 h, 1.3 +/- 1.2 g/dl at 8 h, and 2.1 +/- 0.8 g/dl at 12 h. This increase in alveolar protein concentration was proportional to the volume of liquid removed from the lungs. beta-Adrenergic agonist therapy with terbutaline (10(-5) M mixed with the instilled plasma) doubled the volume of liquid cleared from the lungs over 4 h, and the alveolar protein concentration increased proportionally. However, lung liquid clearance in dogs that were treated with beta-agonists was proportionally (50%) less than in sheep treated with beta-agonists. The slower liquid clearance in dogs compared with sheep cannot be explained by differences in hemodynamics, pulmonary blood flow, anesthesia, mode of ventilation, or alveolar surface area.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sequence of perivascular liquid accumulation in liquid-inflated dog lung lobes

The peribronchovascular interstitium of the lung is a potential space that expands in pulmonary edema with the formation of large liquid cuffs. To study the time course of cuff formation we inflated nine isolated dog lung lobes with liquid to total lung capacity, rapidly froze them in liquid N2 after inflation periods of 1-300 min, then photographed 20 blocks of each lobe at X3 magnification. From the photographs we measured the ratio of cuff area to vessel area for arteries and veins of 0.05-8 mm diam. We found that the cuff-to-vessel area ratio attained a maximum value of 3-4, which was independent of vessel size. However, the first cuffs to reach maximum size were those around vessels of 0.1-0.5 mm diam, whereas cuffs around larger vessels filled more slowly. No cuffs were visible around vessels smaller than 0.1 mm diam. After 45 min cuffs had formed around 99% of all vessels larger than 0.5 mm diam but had formed around only 38% of veins and 91% of arteries of smaller diameter. We simulated the observed rate and pattern of cuff growth using electrical analog models. The filling pattern and model analyses suggest that liquid entered the interstitium from an air space site associated with arteries of approximately 0.1-1.0 mm diam, spread to adjacent sites, and eventually reached the lobe hilum. The estimated perivascular interstitial flow resistance decreased approximately 100-fold with cuff expansion.     

Sequence of interstitial liquid accumulation in liquid-inflated sheep lung lobes

In the initial stages of pulmonary edema, liquid accumulates in the lung interstitium and appears as cuffs around pulmonary vessels. To determine the pattern, rate, and magnitude of cuff formation, we inflated sheep lungs to capacity with liquid (inflation pressure 19 cmH2O) for 3-300 min. After freezing the lobes in liquid N2, we measured perivascular cuff size and total perivascular volume in frozen blocks of each lobe and compared the results with previous measurements in dog lungs. Total cuff volume in sheep lungs reached a maximum value of 5% of air space volume, compared with 9% in dog lungs. In sheep lungs 94% of vessels greater than or equal to 0.5 mm diam and 16% of smaller vessels were surrounded by cuffs. In dog lungs these values were 99 and 47%, respectively. The ratio of cuff area to vessel area reached a maximum of 2.3 in sheep lungs and 3.4 in dog lungs. In an electrical analogue model designed to simulate cuff growth, estimated interstitial resistance to liquid flow was 6-15 times higher than similar estimates in dog lungs. These species differences might be the result of differences in the composition of the interstitial gel or to differences in the mechanical linkage between the lung parenchyma and vessel wall.     

Micropuncture measurement of alveolar liquid pressure in excised dog lung lobes

We have investigated the mechanism of alveolar liquid filling in pulmonary edema. We excised, degassed, and intrabronchially filled 14 dog lung lobes from nine dogs with 75, 150, 225, or 350 ml of 5% albumin solution, and then air inflated the lobes to a constant airway pressure of 25 cmH2O. By use of micropipettes, we punctured subpleural alveoli to measure alveolar liquid pressure by the servo-null technique. Alveolar liquid pressure was constant in all lobes despite differences in lobe liquid volume and averaged 10.6 +/- 1.3 cmH2O. Thus, in all lobes a constant pressure drop of 14.4 cmH2O existed from airway to alveolar liquid across the air-liquid interface. We attribute this finding, on the basis of the Laplace equation, to an air-liquid interface of constant radius in all the lobes. In fact, we calculated from the Laplace equation an air-liquid interface radius which equalled morphological estimates of alveolar radius. We conclude that in the steady state, alveoli that contained liquid have a constant radius of curvature of the air-liquid interface possibly because they are always completely liquid filled.     

Dopamine increases lung liquid clearance during mechanical ventilation

Short-term mechanical ventilation with high tidal volume (HVT) causes mild to moderate lung injury and impairs active Na+ transport and lung liquid clearance in rats. Dopamine (DA) enhances active Na+ transport in normal rat lungs by increasing Na+-K+-ATPase activity in the alveolar epithelium. We examined whether DA would increase alveolar fluid reabsorption in rats ventilated with HVT for 40 min compared with those ventilated with low tidal volume (LVT) and with nonventilated rats. Similar to previous reports, HVT ventilation decreased alveolar fluid reabsorption by ~50% (P < 0.001). DA increased alveolar fluid reabsorption in nonventilated control rats (by ~60%), LVT ventilated rats (by approximately 55%), and HVT ventilated rats (by ~200%). In parallel studies, DA increased Na+-K+-ATPase activity in cultured rat alveolar epithelial type II cells (ATII). Depolymerization of cellular microtubules by colchicine inhibited the effect of DA on HVT ventilated rats as well as on Na+-K+-ATPase activity in ATII cells. Neither DA nor colchicine affected the short-term Na+-K+-ATPase alpha1- and beta1-subunit mRNA steady-state levels or total alpha1- and beta1-subunit protein abundance in ATII cells. Thus we reason that DA improved alveolar fluid reabsorption in rats ventilated with HVT by upregulating the Na+-K+-ATPase function in alveolar epithelial cells.     

Epidermal growth factor increases lung liquid clearance in rat lungs

Epidermal growthfactor (EGF) has been reported to stimulate the proliferation ofepithelial cells and increase Na⁺flux andNa⁺-K⁺-ATPasefunction in alveolar epithelial cell monolayers. Increases inNa⁺-K⁺-ATPasein alveolar type II cells (AT2) have been associated with increasedactive Na⁺ transport and lungedema clearance across the rat alveolar epithelium in a model ofproliferative lung injury. Thus we tested whether administration ofaerosolized EGF to rat lungs would increase activeNa⁺ transport and lung liquidclearance. Sixteen adult Sprague-Dawley male rats were randomized tothree groups. To a group of six rats, an aerosol generated from 20 µgof EGF in saline was delivered to the lungs, to a second group of fiverats only aerosolized saline was delivered, and a third group of fiverats without treatment served as the control. Forty-eight hourspostaerosolization of rat lungs with EGF there was an ~40% increasein active Na⁺ transport and lungliquid clearance compared with control rats, in the absence of changesin²²Na⁺,[³H]mannitol, andalbumin permeabilities. TheNa⁺-K⁺-ATPaseactivity in AT2 cells harvested from these lungs was increased in ratsthat received aerosolized EGF compared with AT2 cells from both controlrats and rats receiving aerosolized saline. These results support thehypothesis that in vivo delivery of EGF aerosols upregulates alveolarepithelialNa⁺-K⁺-ATPaseand increases lung liquid clearance in rats.