Adaptation to hypoxia has attracted much public interest because of its clinical significance. However, hypoxic adaptation in the body is complicated and difficult to fully explore. To explore previously unknown conserved mechanisms and key proteins involved in hypoxic adaptation in different species, we first used a yeast model for mechanistic screening. Further multi-omics analyses in multiple species including yeast, zebrafish and mice revealed that glycerophospholipid metabolism was significantly involved in hypoxic adaptation with up-regulation of lysophospholipid acyltransferase (ALE1) in yeast, a key protein for the formation of dipalmitoyl phosphatidylcholine [DPPC (16:0/16:0)], which is a saturated phosphatidylcholine. Importantly, a mammalian homolog of ALE1, lysophosphatidylcholine acyltransferase 1 (LPCAT1), enhanced DPPC levels at the cell membrane and exhibited the same protective effect in mammalian cells under hypoxic conditions. DPPC supplementation effectively attenuated growth restriction, maintained cell membrane integrity and increased the expression of epidermal growth factor receptor under hypoxic conditions, but unsaturated phosphatidylcholine did not. In agreement with these findings, DPPC treatment could also repair hypoxic injury of intestinal mucosa in mice. Taken together, ALE1/LPCAT1-mediated DPPC formation, a key pathway of glycerophospholipid metabolism, is crucial for cell viability under hypoxic conditions. Moreover, we found that ALE1 was also involved in glycolysis to maintain sufficient survival conditions for yeast. The present study offers a novel approach to understanding lipid metabolism under hypoxia and provides new insights into treating hypoxia-related diseases.

BACKGROUND: Egg-derived peptides are becoming increasingly popular due to their biological activity and non-toxic effects. The egg-derived peptides Arg-Val-Pro-Ser-Leu (RVPSL) and Gln-Ile-Gly-Leu-Phe (QIGLF) display strong angiotensin-converting enzyme inhibitory activity and they can be taken up by intestinal epithelial cells. The interaction of the egg-derived peptides RVPSL and QIGLF with the membrane remains unclear.
RESULTS: The position and structure of the peptides in the membrane were calculated. The maximum density values of RVPSL and QIGLF were 2.27 and 1.22 nm from the center of the 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) membrane, respectively, indicating that peptides penetrated the membrane-water interface and were embedded in the membrane. The interaction of RVPSL and QIGLF with the DPPC membrane did not affect the average area per lipid or the lipid sequence parameters. The thermodynamic parameters ΔH, ΔG, and ΔS of the interaction between the peptide RVPSL with the DPPC membrane were 17.91 kJ mol-1 , -17.63 kJ mol-1 , 187.5 J mol-1 ·k-1 , respectively. The thermodynamic parameters ΔH, ΔG, and ΔS of the interaction between peptide QIGLF with DPPC membrane were 17.10 kJ mol-1 , -17.12 kJ mol-1 , 114.8 J mol-1 ·k-1 , respectively.
CONCLUSIONS: The results indicated that the binding of peptides RVPSL and QIGLF to DPPC is an endothermic, spontaneous, and entropy-driven reaction. The results of the study are relevant to the problem of the low bioavailability of bioactive peptides (BP). © 2023 Society of Chemical Industry.

Background: Acinetobacter baumannii-mediated bacterial pneumonia is a common disease that is harmful to human health. Dipalmitoylphosphatidylcholine (DPPC) is the major lipid component of the pulmonary surfactant (PS) found in the alveolar space; the PS helps to keep surface tension low, which allows for improved oxygen delivery. Resveratrol (RE) is a phytoalexin found in plants that is released in response to injury or infection. The therapeutic effect of Re is limited due to its low solubility and bioavailability. In this study, we report pulmonary delivery of Re-loaded DPPC liposomal large porous microparticles (RDLPMs) for treatment of A. baumannii-induced pneumonia. Methods: Novel RDLPMs were prepared by rotary evaporation and a freeze-drying method in this study. RDLPMs were evaluated by the particle size, electric potential, in vitro release, and particle size distribution. A rat model of A. baumannii-mediated pneumonia was established and used for pharmacodynamic evaluations. Results: The Re-loaded DPPC liposomes (RDLs) consisted of Re/DPPC (1:3, mol/mol) and DPPC/cholesterol (3:1, w/w), with a hydration time of 15 minutes. The RDLs had a high encapsulation efficiency of 69.8% ± 1.6%, a mean size of 191.5 ± 4.5 nm, and a high zeta potential of 12.4 ± 1.5 mV. The RDLPMs were composed of mannitol/large porous microparticles/RDLs (1:4:2, w/w/w) and had a loading efficiency of 2.20% ± 0.24%. The RDLPMs had an aerodynamic diameter (2.73 ± 0.65 μm), a good fluidity (28.30° ± 6.13°), and demonstrated high lung deposition (fine particle fraction = 43.33%). Surprisingly, while penicillin showed better microbial inhibition than the RDLPMs and Re groups in vitro, the RDLPMs were more effective in vivo. Conclusion: The RDLPMs showed good powder properties for pulmonary delivery. The RDLPMs may inhibit the nuclear factor kappa-B pathway and downregulate the expression of cytokines downstream of tumor necrosis factor-α and interleukin-1β. As well as, RDLPMs demonstrated some antibacterial properties against A. baumannii bacteria. Re, when delivered in RDLPMs as a dry powder inhaler, is a promising substitute for antibiotics in the treatment of A. baumannii pneumonia.

Iron oxide nanoparticles (IONPs) are one of the most important components in airborne particulate matter that originally generated from traffic emission, iron ore mining, coal combustion and melting of engine fragments. Once IONPs entered respiratory tract and deposit in the alveoli, they may interact with pulmonary surfactant (PS) that distributed in the alveolar lining. Thereafter, it is necessary to investigate the interaction of inhaled IONPs and PS, which helps the understanding of health risk of respiratory health induced by IONPs. Using dipalmitoyl phosphatidylcholine (DPPC), the major components of PS, as a lipid model, we explored the interaction of DPPC with typical IONPs, Fe3O4 NPs and amino-functionalized analogue (Fe3O4-NH2 NPs). DPPC was readily adsorbed on the surface of both IONPs. Although DPPC corona depressed the cellular uptake of IONPs, IONPs@DPPC complexes caused higher cytotoxicity toward RAW 264.7 macrophages, compared to pristine IONPs. Mechanistic studies have shown that IONPs react with intracellular hydrogen peroxide, which promotes the Fenton reaction, to generate hydroxyl radicals. Iron ions could oxidize lipids to form lipid peroxides, and lipid hydroperoxides will decompose to generate hydroxyl radicals, which further promote cellular oxidative stress, lipid accumulation, foam cell formation, and the release of inflammatory factors. These findings demonstrated the phenomenon of coronal component oxidation, which contributed to IONPs-induced cytotoxicity. This study offered a brand-new toxicological mechanism of IONPs at the molecular level, which is helpful for further understanding the adverse effects of IONPs.

OBJECTIVE: Lung surfactant protects lung tissue and reduces the surface tension in the alveoli during respiration. Particulate matter with an aerodynamic diameter of less than 2.5 μm (PM2.5), which invades primely through inhalation, can deposit on and interact with the surfactant layer, leading to changes in the biophysical and morphological properties of the lung surfactant.
METHODS: Langmuir monolayers of 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) and clinical surfactant Calsurf were investigated with a PM2.5 model injected into the water subphase, which were characterized by surface pressure-area isotherms, Brewster angle microscopy, atomic force microscopy, fluorescent microscopy, and x-ray photoelectron spectroscopy. The binding between DPPC/Calsurf and PM2.5 was studied using isothermal titration calorimetry.
RESULTS: PM2.5 induced the expansion of the monolayers at low surface pressure (п) and film condensation at high п. Aggregation of PM2.5 mainly occurred at the interface of liquid expanded/liquid condensed (LE/LC) phases. PM2.5 led to slimmer and ramified LC domains on DPPC and the reduction of nano-sized condensed domains on Calsurf. Both DPPC and Calsurf showed fast binding with PM2.5 through complex binding modes attributed to the heterogeneity and amphiphilic property of PM2.5. This study improves the fundamental understanding of PM2.5-lung surfactant interaction and shows useful implications of the toxicity of PM2.5 through respiration process.

Background: Acute lung injury is a severe respiratory disorder characterized by overwhelming lung inflammation. Dipalmitoylphosphatidylcholine (DPPC) is the major lipid component of pulmonary surfactant, which here acts as a carrier delivery system for drugs, while also preserving surface tension in the lung. The clinical development of naringenin (NG) is limited by its low solubility and bioavailability. Methods: Novel NG-loaded DPPC phytosomes for dry powder inhalation (NPDPIs) were prepared by solvent evaporation and a freeze-drying method. The particle size, electric potential, in vitro release, and lung deposition were characterized. A rat model of acute lung injury was established and used for pharmacodynamic evaluations. Results: A mixture of NG/DPPC 1:2 (w/w) formed stable phytosomes with the addition of appropriate ethanol. The phytosomes had high complexation efficiency (92.1% ± 1.87%) with NG, a small mean size (150.8 ± 6.9 nm), and a high zeta potential (20.97 ± 0.55 mV). NPDPIs composed of mannitol/DPPC/NG (4:2:1, w/w/w) presented a satisfactory appearance, good fluidity, quick reconstitution to naringenin phytosomes (NGPs), and small (167.2 nm) reconstituted NGPs. The aerodynamic diameter (12.48 μm) and fine particle fraction (23.90%) were suitable for pulmonary delivery by inhalation. The in vivo NPDPIs demonstrated efficacy in a rat model of acute lung injury. NPDPIs significantly inhibited the phosphorylation of P38 in the mitogen-activated protein kinase pathway and suppressed oxidative stress. Surprisingly, the DPPC vehicle exhibited potential effects against acute lung injury by protecting respiratory function. Conclusions: NPDPIs were developed for sustained drug release, promoting pulmonary bioavailability of drug and protecting against acid-induced acute lung injury in rats by pulmonary delivery. NPDPIs are a promising dry powder inhaler for clinical application in acute lung injury.

In this paper, the adsorption behavior of DNA on 1,2-dipalmitoyl-sn-glycero-3- phosphocholine (DPPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) mixed lipid monolayers had been studied at the air-water interface through the surface pressure-area curves (π-A), adsorption curves (π/π0-t), excess mean area (∆Aexc), excess Gibbs free energy (∆Gex) and the atomic force microscopy (AFM). π-A isotherms showed that the curves moved to larger mean molecular area after DNA added into subphase, however, the curves shifted to smaller mean molecular area when the concentration of DNA was higher than 1.2 μg/mL. The result of adsorption curves indicated that DNA molecules were spread by combining with polar head groups of lipids except the concentration of DNA was 0.4 μg/mL. ∆Aexc and ∆Gex demonstrated that DNA enlarged the interval between DPPC and POPC, and the strongest position happened at the concentration of DNA was 1.2 μg/mL. These phenomena might be the steric hindrance between DNA molecules. Morphology of surface observed by AFM was agreement with the results above, which verified our conclusion from a more intuitive aspect. This work provides useful theoretical basis for the development of novel DNA delivery materials.

In this study, the interaction between Lycium barbarum polysaccharide (LBP) and unsaturated 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) or saturated 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was explored using the Langmuir films technique and atomic force microscopy (AFM). Comparing the pure lipid monolayer with the mixed monolayers, the π-A isotherms of the mixed monolayers shifted to larger molecular areas when LBP was added to the subphase. The compression modulus showed that the compressibility of the monolayer films decreased with the addition of LBP. Adsorption curves revealed that the variation in the surface pressure of LBP with POPC was larger than that with DPPC. This phenomenon was verified by the AFM images and the number of each lipid molecule combining with polysaccharide molecules in the mixed monolayer (Ap value), indicating that hydrophobic interactions between LBP and POPC are stronger than those of DPPC. These findings lay the foundation for exploring the pharmacological mechanism of LBP as an in vivo therapeutic.

In this work, the effect of silica nanoparticles (NPs) adding to DPPC monolayer and the interaction between DPPC and silica nanoparticles are studied. Silica nanoparticles are prepared by microemulsion, meanwhile, DMDCS and APTES are used to modify silica NPs to get three types of modified silica NPs. These samples are mixed with DPPC to form mixed monolayer. By using the atomic force microscope (AFM), surface pressure-area and pressure-time isotherms, the effects of different hydrophilic-hydrophobic silica nanoparticles on the interface of lipid monolayer is analyzed. The data shows that the addition of silica nanoparticles changes the phase behavior, the collapse time and the structure of monolayer. Hydrophilic silica NPs decreases the collapse pressure and rigidity of DPPC monolayer, and makes monolayer collapse earlier since the steric hindrance leads to the resistance to compression, while hydrophobic silica NPs have less effect on monolayer in collapse pressure or rigidity but the texture of monolayer, and the addition of hydrophobic NPs causes the appearance of holes in the monolayer. We suppose that there are several possible locations of hydrophobic and hydrophilic silica nanoparticles in the air-water interface, which leads to different effects on the structure and rheological behavior of monolayer. This study can deepen the understanding on how nanoparticles affect human body since industries of nanoparticles on drug delivery, oil recovery and floatation are developing rapidly and getting more and more outside interest on a daily basis.

Cytochrome c (Cyt c) is an essential component of the inner mitochondrial respiratory chain because of its function of transferring electrons. The feature is closely related to the interaction between Cyt c and membrane lipids. We used Langmuir-Blodgett monolayer technique combined with AFM to study the interaction of Cyt c with lipid monolayers at air-buffer interface. In our work, by comparing the mixed Cyt c-anionic (DPPS) and Cyt c-zwitterionic (DPPC/DPPE) monolayers, the adsorption capacity of Cyt c on lipid monolayers is DPPS>DPPE>DPPC, which is attributed to their different headgroup structures. π-A isothermal data show that Cyt c (v=2.5 μL) molecules are at maximum adsorption quantity on lipid monolayer. Moreover, Cyt c molecules would form aggregations and drag some lipids with them into subphase if the protein exceeds the maximum adsorption quantity. π-T curve indicates that it takes more time for Cyt c molecular conformation to rearrange on DPPE monolayer than on DPPC. The compressibility study reveals that the adsorption or intermolecular aggregation of Cyt c molecules on lipid monolayer will change the membrane fluidization. In order to quantitatively estimate Cyt c molecular adsorption properties on lipid monolayers, we fit the experimental isotherm with a simple surface state equation. A theoretical model is also introduced to analyze the liquid expanded (LE) to liquid condensed (LC) phase transition of DPPC monolayer. The results of theoretical analysis are in good agreement with the experiment.