Pulmonary Phospholipids Research Articles

Pulmonary Phospholipid Components as Promising Natural Inhibitors against COVID-19 Mpro; Molecular Docking Analysis Based Study

The ongoing pandemic of COVID-19 caused by the severe acute respiratory syndrome SARS-CoV-2 has become a global crisis. Phospholipids are structural components of mammalian cell membranes that suppress viral attachment to the plasma membrane and subsequent replication in lung cells. Using the molecular docking approach, the inhibitory activity of phosphatidylcholine, dipalmitoylphosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, lysobisphosphatidic acid and sphingomyelin against SARS CoV-2 by targeting main protease (Mpro, PDB code: 6LU7) has been investigated. All phospholipids established excellent binding to Mpro active bocket by forming several H-bonds with the catalytic amino acids Cys145 and His4, as well as various amino acids involved in the bocket. Furthermore, a potent binding affinity is increased from -7.01 to -9.16 kcal/mol compared to compound N3 (N-[(5-methylisoxazol-3-yl)carbonyl]alanyl- L (where L = valyl-N-1-(1R,2Z)-4-(benzyloxy)-4-oxo-1-{[(3R)-2-oxopyrrolidin-3-yl]methyl}but-2- enyl)-L-leucinamide), a peptide linker, inhibitor for Covid-19 main protease. Co-crystalline ligand of enzyme 6LU7 of -9.99 kcal/mol. The sphingomyelin has the same binding affinity to main protease when compared to compound N3. These findings implied that the selected compounds have the potential to be developed as novel SARS-CoV-2 inhibitors. Therefore, improved, well-designed, potent and structurally and pharmacokinetically effective drugs are urgently needed. Further investigations should focus on validating and finalizing effective drugs for COVID-19 beyond preliminary in silico and in vivo screening.

Repeated inhalation exposure of rats to an anionic high molecular weight polymer aerosol: Application of prediction models to better understand pulmonary effects and modes of action

Opposed to the wealth of information available for kinetic lung overload-related effects of poorly-soluble, low-toxicity particles (PSP), only limited information is available on biodegradable high molecular weight (HMW) organic polymers (molecular weight >20,000 Da). It is hypothesized that such types of polymers may exert a somewhat similar volume displacement-related mode of action in alveolar macrophages as PSP; however, with a differing biokinetics of the material retained in the lung. This polyurethane polymer was examined in single and 2-/13-week repeated exposure rat inhalation bioassays. The design of studies was adapted to that commonly applied for PSP. Rats were nose-only exposed for 6h/day for the respective study duration, followed by 1-, 2- and 4-week postexposure periods in the single, 2- and 13-week studies, respectively. While the findings in bronchoalveolar lavage (BAL) and histopathology were consistent with those typical of PSP, they appear to be superimposed by pulmonary phospholipidosis and a much faster reversibility of pulmonary inflammation. Kinetic modeling designed to estimate the accumulated lung burden of biopersistent PSP was also suitable to simulate the overload-dependent outcomes of this biodegradable polymer as long as the faster than normal elimination kinetics was observed and an additional 'void space volume' was added to adjust for the phagocytosed additional fraction of pulmonary phospholipids. The changes observed following repeated inhalation exposure appear to be consistent with a retention-related etiopathology (kinetic overload). In summary, this study did not reveal evidence of any polymer-specific pulmonary irritation or parenchymal injury. Taking all findings into account, 7 mg polymer/m(3) (exposure 6h/day, 5-days/week on 13 consecutive weeks) constitutes the point of departure for lower respiratory tract findings that represent a transitional state from effects attributable to an overload-dependent pulmonary inflammation and phospholipidosis. In regard to extrapulmonary toxicity, no effects were found up to the maximum concentration of 107 mg/m(3) examined.

Body mass related variations in the polar lipid fatty acyl chain composition of the mammalian lung and alveolar surfactant

In nine mammalian species (mouse - cattle: 21.5 g-503 kg) lung total phospholipids (PL), alveolar surfactant phosphatidylcholine (PC) and sphingomyelin (SM) fatty acyl (FA) chain composition was tested relating to body mass (BM) and resting respiratory rate (RRR) associated adaptations. In PL, PC and SM oleic acid (C18:1 n9) provided negative correlations with RRR. Palmitic acid (C16:0) was strongly, positively correlated with RRR in the pulmonary PLs, and myristic (C14:0) acid correlated positively with RRR in the surfactant PCs. In pulmonary PLs negative allometry was found for myristic, palmitic, palmitoleic (C16:1 n7) and docosahexaenoic (C22:6 n3) acids and total saturation, while oleic (C18:1 n9), alpha-linolenic (C18:3 n3) and gondoic (C20:1 n9) acids, total n9 FA s and monounsaturation increased allometrically. In surfactant PC FA s palmitic acid provided negative, while oleic acid and monounsaturation positive allometry; the average FA chain length (ACL) was identical in all species. Surfactant SM FA composition was fully species independent for palmitic and arachidonic acids, total saturation, monounsaturation and ACL. The in vivo lipid peroxidation rate was species independent. The variability of lung PLs was consonant with the "membrane pacemakers theory", while surfactant PC composition was mostly related to RRR.

Time Resolved Studies of Interfacial Reactions of Ozone with Pulmonary Phospholipid Surfactants Using Field Induced Droplet Ionization Mass Spectrometry

Field induced droplet ionization mass spectrometry (FIDI-MS) comprises a soft ionization method to sample ions from the surface of microliter droplets. A pulsed electric field stretches neutral droplets until they develop dual Taylor cones, emitting streams of positively and negatively charged submicrometer droplets in opposite directions, with the desired polarity being directed into a mass spectrometer for analysis. This methodology is employed to study the heterogeneous ozonolysis of 1-palmitoyl-2-oleoyl-sn-phosphatidylglycerol (POPG) at the air-liquid interface in negative ion mode using FIDI mass spectrometry. Our results demonstrate unique characteristics of the heterogeneous reactions at the air-liquid interface. We observe the hydroxyhydroperoxide and the secondary ozonide as major products of POPG ozonolysis in the FIDI-MS spectra. These products are metastable and difficult to observe in the bulk phase, using standard electrospray ionization (ESI) for mass spectrometric analysis. We also present studies of the heterogeneous ozonolysis of a mixture of saturated and unsaturated phospholipids at the air-liquid interface. A mixture of the saturated phospholipid 1,2-dipalmitoyl-sn-phosphatidylglycerol (DPPG) and unsaturated POPG is investigated in negative ion mode using FIDI-MS while a mixture of 1,2-dipalmitoyl-sn-phosphatidylcholine (DPPC) and 1-stearoyl-2-oleoyl-sn-phosphatidylcholine (SOPC) surfactant is studied in positive ion mode. In both cases FIDI-MS shows the saturated and unsaturated pulmonary surfactants form a mixed interfacial layer. Only the unsaturated phospholipid reacts with ozone, forming products that are more hydrophilic than the saturated phospholipid. With extensive ozonolysis only the saturated phospholipid remains at the droplet surface. Combining these experimental observations with the results of computational analysis provides an improved understanding of the interfacial structure and chemistry of a surfactant layer system when subject to oxidative stress.

Allometric scaling of fatty acyl chains in fowl liver, lung and kidney, but not in brain phospholipids

The phospholipid (PL) fatty acyl chain (FA) composition (mol%) was determined in the kidney, liver, lung and brain of 8 avian species ranging in body mass from 150g (Japanese quail, Coturnix coturnix japonica) to 19kg (turkey, Meleagris gallopavo). In all organs except the brain, docosahexaenoic acid (C22:6 n3, DHA) was found to show a negative allometric scaling (allometric exponent: B=-0.18; -0.20 and -0.24, for kidney, liver and lung, respectively). With minor inter-organ differences, smaller birds had more n3 FAs and longer FA chains in the renal, hepatic and pulmonary PLs. Comparing our results with literature data on avian skeletal muscle, liver mitochondria and kidney microsomes and divergent mammalian tissues, the present findings in the kidney, liver and lung PLs seem to be a part of a general relationship termed "membranes as metabolic pacemakers". Marked negative allometric scaling was found furthermore for the tissue malondialdehyde concentrations in all organs except the brain (B=-0.17; -0.13 and -0.05, respectively). In the liver and kidney a strong correlation was found between the tissue MDA and DHA levels, expressing the role of DHA in shaping the allometric properties of membrane lipids.

Metabolic precursors of surfactant disaturated-phosphatidylcholine in preterms with respiratory distress

Our objective was to study the metabolic precursors of surfactant disaturated-phosphatidylcholine (DSPC) in preterm infants with respiratory distress syndrome (RDS) on mechanical ventilation. We performed 46 DSPC kinetic studies in 23 preterms on fat-free parenteral nutrition and mechanical ventilation (birth weight = 1167 +/- 451 g, gestational age = 28.5 +/- 2.0 weeks). Eight infants received a simultaneous intravenous infusion of U(13)C-glucose and [16,16,16](2)H-palmitate, eight infants received U(13)C-glucose and (2)H(2)O, and seven received U(13)C-palmitate and (2)H(2)O. Surfactant DSPC kinetics were calculated from the isotopic enrichments of DSPC-palmitate from sequential tracheal aspirates and its metabolic precursors in plasma or urine. DSPC fractional synthesis rate (FSR) was 17 +/- 11, 21 +/- 16, and 15 +/- 6%/day from glucose, palmitate, and body water, respectively (P = 0.36). DSPC-FSR from U(13)C-glucose and (2)H(2)O were significantly correlated and yielded similar estimates (difference of -0.1 +/- 3%) (P = 0.91). The difference in the 15 infants receiving palmitate versus (2)H(2)O or palmitate versus glucose was +6.0 +/- 12%/day (P = 0.21). There was a significant correlation between DSPC-FSRs from plasma glucose and plasma FFA. The contribution of glucose versus palmitate to DSPC-FSR was 49 +/- 20% versus 51 +/- 20%, respectively. Plasma glucose and FFA showed similar contributions to DSPC-FSR in infants with RDS and fat-free parenteral nutrition. FSRs from (2)H(2)O or glucose were highly correlated.

Reversibility of Pulmonary Abnormalities by Conditional Replacement of Surfactant Protein D (SP-D) in Vivo

Surfactant protein D (SP-D) gene-targeted mice develop severe pulmonary disease associated with emphysema, pulmonary lipidosis, and foamy macrophage infiltrations. To determine the potential reversibility of these abnormalities, transgenic mice were developed in which SP-D was conditionally replaced in the respiratory epithelium of SP-D(-/-) mice. SP-D was not detected in the absence of doxycycline. Treatment with doxycycline after birth restored pulmonary SP-D concentrations and corrected pulmonary pathology at adulthood. When SP-D was replaced in adult SP-D(-/-) mice, alveolar SP-D was restored within 3 days, pulmonary lipid abnormalities were corrected, but emphysema persisted. In corrected adult SP-D(-/-) mice, loss of SP-D caused focal emphysema and pulmonary inflammation but did not cause phospholipid abnormalities characteristic of SP-D(-/-) mice. Thus, abnormalities in surfactant phospholipid homeostasis and alveolar macrophage abnormalities were readily corrected by restoration of SP-D. However, once established, emphysema was not reversed by SP-D. SP-D-dependent processes regulating surfactant lipid homeostasis were disassociated from those mediating emphysema.

A comparison of amniotic fluid fetal pulmonary phospholipids in normal and diabetic pregnancy

Our purpose was to determine whether there are differences in the timing of the appearance of various amniotic fluid fetal pulmonary phospholipids in normal and diabetic pregnancy. A case-control study of 295 subjects with diabetes and 590 control subjects was performed by use of gestational age-matched amniocentesis specimens analyzed for lecithin/sphingomyelin (L/S) ratio, phosphatidylinositol (PI), and phosphatidylglycerol (PG) composition. Diabetic subjects were stratified according to type of diabetes, degree of blood glucose control, and birth percentile of the neonate. There was no difference in L/S ratios over gestational age by type of diabetes or quality of glycemic control. Women with preexisting diabetes had significantly higher PI levels at 33 to 35 weeks' gestation, which became similar to levels of control subjects after 36 weeks, whereas patients with gestational diabetes mellitus and control subjects had similar PI levels throughout. In diabetic subjects, the onset of production of PG was delayed from 35.9 +/- 1.1 weeks (controls) to 38.7 +/- 0.9 weeks (overt diabetics) and 37.3 +/- 1.0 weeks for gestational diabetes mellitus (P <.001). The delay in PG synthesis was not related to infant sex, level of maternal glucose control, or fetal macrosomia. Fetal pulmonary maturation, as evidenced by the onset of PG production in the amniotic fluid, is delayed in diabetic pregnancy by 1 to 1.5 weeks. This delay appears to be associated with an early and sustained elevation in amniotic fluid PI levels at 32 to 34 weeks.

Surfactant phospholipids in the bronchoalveolar lavage fluid (BALF) of children who develop acute lung injury (ALI)

Introduction Children with ALI represent 5% of admissions to our Paediatric Intensive Care Unit [1]. Although there has been much research in adults with ALI, little is known about the pathophysiology in children [2]. More specifically there is a paucity of information on the pulmonary phospholipid (PL) changes during ALI children. Methods All children without pre-existing lung pathology who developed ALI (defined as PaO2/FiO2 Results Over 9 months, 40(8.8%) children of 452 admissions developed ALI. Ten of 26 eligible children were enrolled in the study; six parents declined consent and 10 children were either too unstable for BALF collection or were not recruited in time. In the study group, the dipalmitoyl PC (DPPC) content in BALF decreased from control values (42.8 ± 6.5% of total PC) to a minimum of 23.1 ± 11.9% (P < 0.01) between days 2–3, but increased to 37.8 ± 4.8% pre-extubation. These changes were accompanied by reciprocal increases in the concentrations of monounsaturated PC species characteristic of inflammatory cells. Conclusion Changes in the PC profile of BALF samples in children with ALI indicate that there is an increase in the cellular breakdown products. We speculate that this alteration in the PC profile with a lowering of DPPC may contribute to the disease process of ALI in children.

Activity of Pulmonary Surfactant Protein-D (SP-D) in Vivo Is Dependent on Oligomeric Structure

Pulmonary surfactant protein-D (SP-D) is a member of the collectin family of C-type lectins that is synthesized in many tissues including respiratory epithelial cells in the lung. SP-D is assembled predominantly as dodecamers consisting of four homotrimeric subunits each. Association of these subunits is stabilized by interchain disulfide bonds involving two conserved amino-terminal cysteine residues (Cys-15 and Cys-20). Mutant recombinant rat SP-D lacking these residues (RrSP-Dser15/20) is secreted in cell culture as trimeric subunits rather than as dodecamers. In this study, transgenic mice that express this mutant were generated to elucidate the functional importance of SP-D oligomerization in vivo. Expression of RrSP-Dser15/20 failed to correct the pulmonary phospholipid accumulation and emphysema characteristic of SP-D null (mSP-D-/-) mice. Expression of high concentrations of the mutant protein in wild-type mice reduced the abundance of disulfide cross-linked oligomers of endogenous SP-D in the bronchoalveolar lavage fluid and demonstrated a phenotype that partially overlapped with that of the SP-D-/- mice; the animals developed emphysema and foamy macrophages without the associated abnormalities in alveolar phospholipids typical of SP-D-/- mice. Development of foamy macrophages in SP-D-deficient mice is not secondary to the increased abundance of surfactant phospholipids. Disulfide cross-linked SP-D oligomers are required for the regulation of surfactant phospholipid homeostasis and the prevention of emphysema and foamy macrophages in vivo.

Occurrence of Oxidized Metabolites of Arachidonic Acid Esterified to Phospholipids in Murine Lung Tissue

Isolation and characterization of murine pulmonary phospholipids revealed the normal occurrence of 10 isobaric eicosanoids corresponding to the incorporation of one oxygen atom into the arachidonate esterified to glycerophospholipids. Lungs from mice were removed and lipids were extracted and then separated into free carboxylic acid and phospholipids. Phospholipids were hydrolyzed to yield the free carboxylic acids prior to analysis. Reverse-phase HPLC and electrospray tandem mass spectrometry were used to identify and quantitate six monohydroxyeicosatetraenoic (HETE) and four epoxyeicosatetraenoic (EET) acid regioisomers using d8-HETE as internal standard. HETEs esterified to phospholipids were found to increase following intratracheal administration of tBuOOH (36 mg/kg), but not the levels of esterified EETs. Chiral analysis of esterified 15-HETE revealed an R/S ratio of 0.96, suggesting operation of a free radical mechanism responsible for generation of this monohydroxy arachidonate phospholipid, and this enantiomeric ratio was 1.10 following treatment of the mouse lung with tBuOOH. These results are consistent with a free-radical-based mechanism of oxidation of pulmonary glycerophospholipids containing arachidonate.

Antenatal triiodothyronine improves neonatal pulmonary function in preterm lambs.

To characterize 1) pulmonary gas exchange, 2) pulmonary function, 3) lung fluid and tissue phospholipid content, and 4) thyroid hormone in the premature lamb (0.85 of term) after intra-amniotic administration of 100 micrograms of triiodothyronine (T3) 2 weeks before delivery. Nine fetal lambs were given 100 micrograms of intra-amniotic T3 under ultrasound guidance at 112 +/- 1 days' gestation and delivered at 126 +/- 1 days (term = 149 days). Five saline-injected animals served as controls. Arterial blood gases, pulmonary mechanics, and lung volumes were compared between groups for 1 hour after delivery. At delivery, tracheal fluid and blood was taken for T3, and thyroxine (T4) levels. Tracheal fluid and lung tissues were assayed for total phosphorus and disaturated phosphatidylcholine. Triiodothyronine-treated lambs had significantly higher mean arterial pH and lower PCO2 than controls (P < .05) with a trend toward higher mean PO2. The dynamic lung compliance was increased by 54% with a 40% proportional increase in tidal volume and minute ventilation in the T3-treated group (P < .05). Functional residual capacity increased 69% (P < .05) without a change in specific compliance. The tracheal fluid and pulmonary phospholipids and tracheal fluid and plasma T3 and T4 levels were not different between the two groups. A single 100 micrograms dose of antenatal T3 significantly improves neonatal gas exchange and lung compliance. The improvement in lung function was not accompanied by an increase in pulmonary surfactant production. It is inferred that T3 improved lung function via accelerated structural development of the lung with an alternative possible effect on parenchymal connective tissue matrix.

Pulmonary phospholipid saturation increases with glucocorticoid receptor mRNAs in late gestation but not due to labor in the fetal rhesus monkey

We report here changes in abundance of pulmonary cytosolic phospholipase A2 (cPLA2) and glucocorticoid receptor (GR) mRNAs and phospholipid (PL) fatty acids (FA) with gestational age, in either spontaneous or androstenedione-induced premature labor for 15 fetal rhesus monkeys and 1 neonate. Pulmonary RNA and lipids were extracted from the lungs of the rhesus monkeys, gestational ages ranging from 140 days to term. Northern hybridization analysis was performed for both cPLA2 and GR mRNAs. The results demonstrated the transcript of the cPLA2 constantly expressed and that of GR increased gradually in the lungs during the gestational stages studied. Phosphatidylcholine (PC), phosphatidylglycerol (PG) and phosphatidylinositol (PI) were separated by thin layer chromatography and the fatty acid composition in these PL was quantified by gas chromatography (GC). Regression analysis demonstrated that the expression of GR, mRNAs increased significantly with gestational age (P < 0.01). Furthermore, the level of GR mRNA is significantly correlated with that of cPLA2, with Pearson correlation coefficient of 0.83 (P < 0.01). The PC palmitate percentage increased significantly with gestational age (P < 0.01), while the PC unsaturation index and total polyunsaturated FA decreased during late gestation (P < 0.01). Changes in PG FA with gestational age were similar to those in PC, while PI FA did not change with gestational age. There was no effect of either spontaneous or induced labor on mRNA levels of cPLA2 and GR or the FA composition of pulmonary PL studied. We conclude that the pulmonary PL FA compositions are under developmental control.

Effects of maternal dexamethasone therapy on fetal lung development in the rhesus monkey.

A large body of evidence demonstrates that antenatal glucocorticoids can accelerate fetal lung maturation. The purpose of this study was to delineate the optimal dose of dexamethasone and to determine whether a single- or multiple-injection regimen of the same total dexamethasone dosage was more effective in accelerating pulmonary development. Pregnant rhesus monkeys were injected with varying doses of dexamethasone or vehicle as either a single-bolus injection or as four separate injections, each spaced 12 hours apart. All maternal treatments were begun exactly 72 hours prior to delivery, and all fetuses were delivered by cesarean section at 135 +/- 1 days gestation. When a single injection of dexamethasone was used, fetal liver weight increased in a dose-related fashion. Fetal and maternal cortisol and fetal blood glucose concentrations were also influenced by increasing dexamethasone dosages. Fetal pulmonary phospholipids, however, were unchanged at all steroid doses examined. Multiple injections of dexamethasone generally produced more pronounced effects, even though the total dexamethasone dose remained the same. Thus, liver weight and fetal and maternal cortisol, glucose, and insulin levels were significantly influenced by the multiple administration of dexamethasone. In addition, total lung phosphatidylcholine, surfactant phosphatidylcholine, the surfactant-phosphatidylcholine-to-total-phosphatidylcholine ratio, and the surfactant-disaturated-phosphatidylcholine-to-total-lung-disaturated- phosphatidylcholine ratio were elevated after the multiple-injection regimen. A total dose of 0.5 mg dexamethasone/kg maternal body weight given in four separate injections appeared to produce the most beneficial results. These data suggest that low-dose, antenatal glucocorticoid treatment can effectively accelerate the biochemical maturation of the fetal lung.

Acute silica toxicity: attenuation by amiodarone-induced pulmonary phospholipidosis.

Exposure to the toxic mineral dust silica has been shown to produce an acute inflammatory response in the lungs of both humans and laboratory animals. Coating silica with phospholipids reduces its toxicity when studied with in vitro systems. The drug amiodarone increases phospholipid within the cells, airways, and alveoli of the lungs. This increase in phospholipid is due to amiodarone's ability to inhibit phospholipase activity within alveolar macrophages (AMs) and whole lung. The purpose of this study was to determine whether the amiodarone-induced increase in pulmonary phospholipid would protect the lungs from acute damage caused by the intratracheal instillation of silica. Treatment of male Fischer 344 rats with amiodarone for 14 days caused an increase in phospholipid content in bronchoalveolar lavage fluid and AMs compared to vehicle-treated controls. The rats were then instilled with silica or saline vehicle. At both 1 and 14 days after silica exposure, pulmonary phospholipidosis was associated with a marked reduction in acute silica-induced pulmonary damage as assessed by biochemical parameters in bronchoalveolar lavage fluid, however, the influx of neutrophils into the airspaces was not reduced. Four times more phospholipid was bound to the silica recovered from amiodarone-treated rats compared to controls. The results of these in vivo experiments indicate that pulmonary phospholipidosis attenuates the acute damage associated with the intratracheal instillation of silica in rats. By using an in vitro cell culture system, we demonstrated that, in contrast to control AMs, phospholipidotic AMs were significantly more resistant to the cytotoxicity of surfactant-coated silica.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Silica Toxicity: Attenuation by Amiodarone-Induced Pulmonary Phospholipidosis

Exposure to the toxic mineral dust silica has been shown to produce an acute inflammatory response in the lungs of both humans and laboratory animals. Coating silica with phospholipids reduces its toxicity when studied with in vitro systems. The drug amiodarone increases phospholipid within the cells, airways, and alveoli of the lungs. This increase in phospholipid is due to amiodarone's ability to inhibit phospholipase activity within alveolar macrophages (AMs) and whole lung. The purpose of this study was to determine whether the amiodarone-induced increase in pulmonary phospholipid would protect the lungs from acute damage caused by the intratracheal instillation of silica. Treatment of male Fischer 344 rats with amiodarone for 14 days caused an increase in phospholipid content in bronchoalveolar lavage fluid and AMs compared to vehicle-treated controls. The rats were then instilled with silica or saline vehicle. At both 1 and 14 days after silica exposure, pulmonary phospholipidosis was associated with a marked reduction in acute silica-induced pulmonary damage as assessed by biochemical parameters in bronchoalveolar lavage fluid, however, the influx of neutrophils into the airspaces was not reduced. Four times more phospholipid was bound to the silica recovered from amiodarone-treated rats compared to controls. The results of these in vivo experiments indicate that pulmonary phospholipidosis attenuates the acute damage associated with the intratracheal instillation of silica in rats. By using an in vitro cell culture system, we demonstrated that, in contrast to control AMs, phospholipidotic AMs were significantly more resistant to the cytotoxicity of surfactant-coated silica.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of metabolic control of pregnant diabetics on fetal lung maturity.

We studied the relationship between the metabolic control of pregnant diabetics and fetal lung maturity. In 31 diabetic and 20 normal pregnancies we analysed phospholipids in amniotic fluid and glycaemic control parameters. There were no differences in amniotic fluid lecithin/sphingomyelin (L/S) and phosphatidylinositol/sphingomyelin (PI/S) ratios between pregnant diabetics with strict metabolic control and the control group. Pregnant diabetics with poor metabolic control had significantly different L/S and PI/S ratios than the normal pregnant women. Six women in this group of poorly controlled diabetics had mature surfactant in amniotic fluid (L/S > or = 2.7 and presence of phosphatidylglycerol) at 32-34 weeks of amenorrhea; the difference was significant (p < 0.05) with respect to the control group. These six women had recurrent hypoglycaemic episodes (6.4 +/- 1.3 episodes/week) at 14-27 weeks of amenorrhea. Our findings suggest that hypoglycaemic stress on the fetus could disturb fetal synthesis of pulmonary phospholipids.

High-performance liquid chromatographic analysis of lung phospholipids and their precursors in the offspring of diabetic rats

Pulmonary phospholipids and their precursors and metabolites were assayed in the offspring of streptozotocin-induced diabetic rats at 19 and 21 days gestation and at 2 days after birth by two unique methods that employ high performance liquid chromatography combined with automated phosphorus analysis. In general, lung phospholipids were not different between offspring of control versus diabetic mothers. Levels of phosphatidylglycerol, however, were decreased in the offspring of diabetics. Lysophosphatidylcholine appears to be increased in the lungs of offspring of diabetic mothers, suggesting that maternal diabetes is associated with alterations in the remodeling of phosphatidylcholine.

Relationships among surfactant fraction lipids, proteins and biophysical properties in the developing rat lung

Lung development is associated with increases in specific phospholipids and proteins that function as critical pulmonary surfactant components. Attempts to characterize the pattern of surfactant development in fetal rat lungs have been hampered by the lack of a micromethod which will permit quantitative isolation of surface active components from small tissue specimens. As part of studies designed to elucidate the metabolic regulation of lung development in the rat, we developed sucrose density gradient centrifugation procedures to separate pulmonary phospholipids and proteins into a presumed surfactant (S) fraction and a residual (R) fraction. Electron microscopy of S pellets from mature fetuses identified predominant lamellar bodies and minimal contamination; incubation with 5 mM CaCl2 induced the appearance of tubular myelin figures, implying functional potential. This was confirmed by demonstrating low surface tension (less than 1 dyn/cm) in S, but not R, fractions at term gestation (21.5 days) and in 1-day-old neonatal lung isolates, based on dynamic measurements using the oscillating bubble technique. Surface activity was also high in the S pellets from fetuses at 20.5 days of gestation; however, at 19.5 days, minimum surface tension values of at least 19 dyne/cm were seen. These results correlated directly with biochemical analyses which indicated striking increases in three surfactant-associated proteins (SP-A, SP-B, and SP-C) after 19.5 days of gestation; a finding in agreement with previously reported data on the developmental increase of disaturated phosphatidylcholine in fetal rat lung. We conclude that isolation of S fraction components is valuable for demonstrating maturation of the fetal rat lung and may provide a useful tool for the study of regulatory mechanisms influencing surfactant production and function.

The preterm rat: a model for studies of acute and chronic neonatal lung disease.

A preterm rat model has been developed for studies of acute and chronic neonatal lung disease. Premature delivery 24 h before the normal time of delivery is associated with immature pulmonary phospholipid and antioxidant enzyme profiles. The premature lung is more fragile and ruptures at a lower lung vol (172 +/- 8 microL) than the full-term fetal lung (259 +/- 14 microL). Only 7% of premature lungs will float in liquid, after inflation to 85% of the rupture vol, compared with 87% of term fetal lungs. This lung immaturity was reflected in a survival rate of only 6% by 36 h after delivery if the preterm pups were placed in air, which increased to 47% when they were placed in greater than 95% oxygen. Though greater than 95% oxygen enhanced survival of preterm pups during the 1st wk of life, these survivors had a 50% mortality during the 2nd wk of exposure to greater than 95% oxygen. The preterm pup will tolerate intraperitoneal injection of antioxidant enzymes entrapped in liposomes and has a better retention of these liposomes in the lung compared with the term pup. We conclude that the preterm rat is a suitable model for studies of acute and chronic neonatal lung disease.