Abstract
We studied the mechanisms by which C16:0 lysophosphatidylcholine (LPC) and albumin inhibit the surface activity of calf lung surfactant extract (CLSE) by using a pulsating bubble apparatus with a specialized hypophase exchange system, plus adsorption and Wilhelmy balance measurements. In the absence of inhibitors, CLSE (1 mg phospholipid/mL) reached minimum surface tension (γmin) < 1 mN/m within 5 min of bubble pulsation at 20 cycles/min at 37°C. Mixtures of CLSE:LPC had impaired surface activity depending on LPC content: γmin was raised to 5 mN/m by 14 wt % LPC, to 15 mN/m by 25-30 wt % LPC, and to >20 mN/m (67 wt % LPC), even at high CLSE concentrations (3 and 6 mg phospholipid/mL). In contrast, inhibition of CLSE by albumin was more easily abolished when surfactant concentration was raised. Mixtures of albumin (3 mg/mL) and CLSE (1 mg phospholipid/mL) had γmin >20 mN/m, but normal values of γmin < 1 mN/m were reached at higher CLSE concentration (3 mg phospholipid/mL) even when albumin concentration was increased 8-fold to 24 mg/mL. In hypophase exchange studies, LPC, but not albumin, was able to penetrate preformed CLSE surface films and raise γmin. CLSE surface films with γmin < 1 mN/m were isolated by an initial hypophase exchange with saline, and a second exchange with an LPC- containing hypophase raised γmin to ∼10 mN/m. CLSE surface films retained the ability to reach γmin < 1 mN/m in analogous hypophase exchange studies with albumin. The ability of LPC to penetrate surface films of CLSE, although albumin could not, was also demonstrated in absorption experiments in a Teflon dish, where diffusion was minimized by subphase stirring. Wilhelmy balance experiments also demonstrated that LPC could mix and interact with CLSE or dipalmitoyl phosphatidylcholine in solvent-spread surface films. The ability of LPC or other cell membrane lipids to penetrate interfacial films and raise γmin even at high surfactant concentration may increase their inhibitory actions during acute lung injury.
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Abbreviations
- ARDS:
-
acute respiratory distress syndrome
- BAL:
-
bronchoalveolar lavage
- BSA:
-
bovine serum albumin
- CLSE:
-
calf lung surfactant extract
- DPPC:
-
1,2-dipalmitoyl-sn-3-phosphocholine dipalmitoyl phosphatidylcholine
- HEPES:
-
HEPES standard buffer
- LPC:
-
C16:0 lysophosphatidylcholine
- γmin:
-
minimum surface tension
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Supported by National Institutes of Health Grant HL-56176.
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Holm, B., Wang, Z. & Notter, R. Multiple Mechanisms of Lung Surfactant Inhibition. Pediatr Res 46, 85–93 (1999). https://doi.org/10.1203/00006450-199907000-00015
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DOI: https://doi.org/10.1203/00006450-199907000-00015
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