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Ca2(+)-dependent synthesis of prostaglandin I2 and mobilization of arachidonic acid from phospholipids in cultured endothelial cells permeabilized with saponin.

The feasibility of using saponin as a permeabilization agent to study the effect of free Ca2+ concentration ([Ca2+]f) on prostaglandin I2 (PGI2) synthesis and mobilization of arachidonic acid from membrane phospholipids was investigated in cultured bovine pulmonary artery endothelial cells (BPAEC). Treatment of BPAEC with 20 micrograms/ml saponin caused selective permeabilization of the plasma membrane as determined by measurements of the release of lactate dehydrogenase and beta-hexosaminidase. In cells prelabeled with [3H]arachidonic acid for 22 h, permeabilization with 20 micrograms/ml saponin induced PGI2 synthesis and release of [3H]arachidonic acid from membrane phospholipids. These effects were dependent upon [Ca2+]f in the range 72 nM to 5 microM. Release of [3H]arachidonic acid from phospholipid classes was determined in suspensions of BPAEC prelabeled with [3H]arachidonic acid and permeabilized with 20 micrograms/ml saponin. At [Ca2+]f optimal for PGI2 synthesis, 16.2% of the total incorporated [3H]arachidonic acid was released from phosphatidylinositol (3.4%), phosphatidylethanolamine (3.5%) and phosphatidylcholine (9.3%). The time course and dependence upon [Ca2+]f of [3H]arachidonic acid release from phospholipids correlated with PGI2 synthesis. The amount of PGI2 synthesized in permeabilized BPAEC was similar to that in cell cultures treated with the calcium ionophore A23187. In comparison, however, PGI2 synthesis induced by A23187 was associated with less release of [3H]arachidonic acid from membrane phospholipids, e.g., 2.3% versus 16.2%. The greater loss of [3H]arachidonic acid from phospholipids in saponin-permeabilized BPAEC was most likely due to the loss of cell integrity and/or nonspecific effects of the detergent on phospholipases. Despite these limitations, the Ca2+ dependence observed for PGI2 synthesis and [3H]arachidonic acid mobilization suggest that saponin-permeabilization may provide a useful system for studies of the intracellular events triggered by the rise in intracellular Ca2+ which culminate in PGI2 synthesis.

6-Ketoprostaglandin F1 alpha↗

Inhibition of interferon gamma-induced macrophage microbicidal activity against Leishmania major by liposomes: inhibition is dependent upon composition of phospholipid headgroups and fatty acids.

Multilamellar liposomes of phosphatidylcholine and phosphatidylserine at a 7:3 molar ratio significantly inhibited activation of murine resident peritoneal macrophages by recombinant murine interferon-gamma for cytotoxicity against amastigotes of the protozoan parasite Leishmania major; other macrophage effector functions, such as particle phagocytosis or tumoricidal activity, were unaffected. This inhibition was not due to direct toxic effects of liposomes against parasite or macrophage, was fully reversible, and was directed at one or more early events in macrophage-LK interactions which ultimately induce microbicidal activity. Liposomes containing some natural phospholipids (phosphatidylserine, phosphatidylethanolamine, phosphatidic acid or diphosphatidyl glycerol), but not phosphatidylcholine, phosphatidylglycerol, or several synthetic saturated phospholipids, prevented the induction of macrophage microbicidal activity. Inhibition by liposomes of various composition was not related to the efficiency with which these vesicles were ingested by macrophages. Inhibitory activity was directly influenced by changes in the phospholipid head group, as well as by the number of unsaturated bonds in phospholipid fatty acids: for a given phospholipid in liposomes, inhibition was directly related to the number of unsaturated bonds among the fatty acids. These data support a role for phospholipids in postbinding regulation of macrophage activation and add to our understanding of how liposome delivery systems can be designed to avoid potential microbicidal suppressive effects.

Animals↗

The effects of serum albumin and phospholipid on sterol excretion in tissue culture cells.

Serum albumin was as effective as whole serum or alpha-globulins in facilitating sterol release from strain L mouse fibroblasts. Commercial bovine serum albumin preparations, however, had markedly different absolute effects in this regard. These differences were attributable to the variation in phospholipid content of these products. All but one of these albumins enhanced sterol release when supplemented with phospholipid. The exception was fatty acid-poor albumin which contained an adequate amount of phospholipid. Among the phospholipids examined, lecithin proved to be most effective, while phosphatidylethanolamine had little potentiating influence. As the unsaturation of the test lecithins increased, enhancement of sterol release decreased. The potentiating effect of the phospholipid was in turn dependent on the protein used, since the phenomenon was not observed with non-serum proteins like ovalbumin or with non-transport serum proteins such as gamma-globulins. The results of these studies raise the possibility that serum albumin together with phospholipid can play an important role in sterol release in tissue culture cells.

Biological Transport↗

Tissue distribution of EDTA encapsulated within liposomes containing glycolipids or brain phospholipids.

Multilameller liposomes were prepared with various asialoglycolipids, gangliosides, sialic acid, or brain phospholipids in the liposome membrane and with ethylenediaminetetraacetic acid (EDTA) encapsulated in the aqueous compartments. The liposomes containing glycolipids or sialic acid were prepared from a mixture of phosphatidylcholine, cholesterol, and one of the following test substances: galactocerebroside, glucocerebroside, galactocerebroside sulfate, mixed gangliosides, monosialoganglioside GM1, monosialoganglioside GM2, monosialoganglioside GM3, disialoganglioside GD1a, or sialic acid. The liposomes containing brain phospholipids were mixtures of either sphingomyelin and cholesterol or a brain total phospholipid extract and cholesterol. Distributions of 14C-labeled EDTA were determined in mouse tissues from 15 min to 6 h or 12 h after a single injection of liposome preparation. Liver uptake up encapsulated EDTA was lowest from all liposome preparations containing sialic acid or sialogangliosides, regardless of the amount of sialic acid moiety present or the identity of the particular ganglioside; highest uptake of encapsulated EDTA by liver was from liposomes containing galactocerebroside or brain phospholipids. Lungs and brain took up the largest amounts of EDTA from liposomes containing sphingomyelin and lesser amounts from liposomes containing GD1a. Use of mouse brain phospholipid extract to prepare liposomes did not increase uptake of encapsulated EDTA by the brain. EDTA in liposomes containing monosialogangliosides, brain phospholipids, galactocerebroside, or sialic acid was taken up well by spleen and marrow. Highest thymus uptake of encapsulated EDTA was from liposomes containing GD1a. These results demonstrate that inclusion of sialogangliosides in liposome membranes decreases uptake of liposomes by liver, thus making direction of encapsulated drugs to other organs more feasible. Liposomes containing glycolipids also have potential uses as probes of cell surface receptors.

Animals↗

Phospholipid and fatty acid composition of erythrocyte membrane from wild Japanese serow (Capricornis crispus).

The phospholipid composition and fatty acid composition of the individual phospholipids were determined in erythrocyte membrane of wild Japanese serow, Capricornis crispus, and compared with those of Japanese cattle. Sphingomyelin (SM) contributed more than 50% to the total phospholipids, with only 3% phosphatidylcholine, 30% phosphatidylethanolamine and 11% phosphatidylserine. This phospholipid composition and ratio of phospholipid to protein in erythrocyte membrane of wild serow were quite similar to those of Japanese cattle. However, marked differences in fatty acid composition were found, especially in lignoceric acid 24:0 and nervonic acid 24:1 of sphingomyelin which were major constituents (approximately 60%) of that phospholipid.

Animals↗

Control of gastric mucus phospholipid content and composition by cholinergic and adrenergic mediators.

1. The secretion of choline-containing phospholipids by gastric mucosal cells in response to neural mediators was investigated using beta-adrenergic and cholinergic agents. 2. A 2.7-fold increase in phospholipid secretion occurred with isoproterenol, while pilocarpine evoked 1.4-fold increase and the effects were inhibited by the respective antagonists. 3. The phospholipid secretory responses were stimulated by dibutyryl-cAMP and phorbol myristate acetate (PMA), but not by 4 alpha-phorbol-12,13-didecanoate which does not activate protein kinase C. The effects of dibutyryl-cAMP and PMA were additive, the the PMA induced phospholipid secretion was inhibited by a protein kinase C inhibitor, tetracaine. 4. The phospholipids secreted in response to isoproterenol showed a 2.1-fold decrease in lysophosphatidylcholine, while those secreted in response to pilocarpine were enriched 2.3-fold in lysophosphatidylcholine, and 1.5-fold in sphingomyelin, and showed 23% lower content of phosphatidylcholine. 5. The results suggest that cholinergic and beta-adrenergic mediators participate in defining the gastric mucus phospholipid content and composition, and hence influence the mucosal protective capability.

Adrenergic beta-Agonists↗

Thin-layer chromatography and high-performance liquid chromatography for the assay of fatty acid compositions of individual phospholipids in platelets from non-insulin-dependent diabetes mellitus patients: effect of eicosapentaenoic acid ethyl ester administration.

Eight major phospholipids were separated by a TLC method with a one-dimensional developing system without any pretreatment of the plate and the fatty acids incorporated into each phospholipid class were analysed by an improved HPLC method with a simple elution system, which has advantages with respect to resolution and analysis time. The fatty acid compositions of individual phospholipids in platelets were investigated following administration of ethyl cis-5,8,11,14,17-eicosapentaenoate for more than 13 weeks to patients with non-insulin-dependent diabetes mellitus. The cis-5,8,11,14,17-eicosapentaenoic acid compositions of all phospholipid classes were significantly increased with decreasing platelet aggregation rates after the administration. These results suggested that the present method provides the complete separation of individual phospholipids in sufficient amounts to allow fatty acid analysis on the isolated phospholipid moieties.

Blood Platelets↗

Insulin inhibits changes in the phospholipid profiles in sciatic nerves from streptozocin-induced diabetic rats: a phosphorus-31 magnetic resonance study.

Sciatic nerve phospholipids obtained from insulin-treated streptozocin-induced diabetic, non-treated streptozocin-induced diabetic, and healthy, control male Sprague-Dawley rats after eighteen weeks of diabetes were studied by 31P NMR spectrometry. Eleven phospholipids resonances were identified as follows: Phosphatidic acid (Chemical shift, 0.30 ppm), dihydrosphingomyelin (0.13 ppm), ethanolamine plasmalogen (0.07 ppm), phosphatidylethanolamine (0.03 ppm), phosphatidylserine (-0.05 ppm), sphingomyelin (-0.09 ppm), lysophosphatidylcholine (-0.28 ppm), phosphatidylinositol (-0.30 ppm), alkylacylglycerophosphorylcholine (-0.78 ppm), choline plasmalogen (-0.80 ppm), and phosphatidylcholine (-0.84 ppm). Diabetic rats showed that phosphatidylcholine was significantly elevated (p < 0.05), and ethanolamine plasmalogen and choline plasmalogen were significantly lower when compared with both control and insulin treated rats. The choline ratio (choline-containing phospholipids over noncholine phospholipids) was significantly elevated in the diabetic group, when compared with both control and insulin-treated groups. The ethanolamine ratio (ethanolamine-containing phospholipids over nonethanolamine phospholipids) and the ratio of the ethanolamine ratio over the choline ratio, was significantly elevated in the control and the insulin-treated groups when compared with the diabetic group. The presence of phosphatidic acid and the significance in phosphatidylcholine and ethanolamine plasmalogen, suggested that insulin had a role in the phosphatidylcholine metabolism in the rat nerve.

Animals↗

Gas chromatography-mass spectrometry method for determination of phospholipid peroxides; II. Transesterification to form pentafluorobenzyl esters and detection with picogram sensitivity.

The purpose of this study is to develop a higher sensitivity method to detect phospholipid peroxides in tissues to more effectively investigate the role of lipid peroxidation in pathology. We recently presented a gas chromatography-mass spectrometry (GC-MS) method for identification and measurement of phospholipid peroxides at the 10 ng level (van Kuijk, et al. J. Free Rad. Biol. Med., 1:215-225, 1985) based on transesterification to form fatty acid methyl ester derivatives. While this method is quite useful under some circumstances, more sensitivity was desirable to analyse trace amounts of lipid peroxidation products in small samples of various tissues. We present a method to prepare pentafluorobenzyl esters of fatty acids and hydroxy fatty acids directly from phospholipids by transesterification at room temperature or from triglycerides at 60 degrees C. Oxidized lipids are extracted into dichloromethane and peroxide functions are reduced with sodium borohydride. Transesterification of phospholipids is carried out in dichloromethane with 20% (v/v) pentafluorobenzyl alcohol, 1% (w/v) potassium tert-butoxide and 0.2 N (m-trifluoromethyl phenyl) trimethyl ammonium hydroxide at room temperature for 30 min. Pentafluorobenzyl esters are purified on open silica columns, O-trimethyl silyl derivatives of the alcohol functions are formed, and GC-MS is carried out with negative ion chemical ionization detection. The single or double oxidation products of oleate, linoleate, arachidonate, and docosahexaenoate were detected with 10 pg sensitivity in oxidized rat retina phospholipids or synthetic phospholipids, by negative ion chemical ionization with specific ion monitoring of carboxylate anions.

Borohydrides↗

Comparable interaction of doxorubicin with various acidic phospholipids results in changes of lipid order and dynamics.

We have characterized the interaction of the antitumor drug doxorubicin with model membranes of the anionic phospholipids dioleoylphosphatidic acid (DOPA), dioleoylphosphatidylserine (DOPS), cardiolipin and dioleoylphosphatidylglycerol (DOPG) as compared to the zwitterionic dioleoylphosphatidylcholine (DOPC) or dioleoylphosphatidylethanolamine (DOPE). The saturating binding levels were: 2.4 (DOPA), 1.3 (cardiolipin), 1.5 (DOPS, DOPG) and 0.02 (DOPC) doxorubicin per lipid phosphorus (mol/mol). The half-saturating free drug concentrations were comparable for DOPA, cardiolipin, DOPS and DOPG: 20, 16, 35 and 18 microM, respectively. Doxorubicin fluorescence revealed the simultaneous existence of at least two populations of bound drug in the various anionic phospholipids: (1) fluorescent molecules with chromophores that reside between the lipid molecules and (2) above 0.01-0.02 doxorubicin bound per lipid phosphorus: non-fluorescent drug-stacks that are closer to the aqueous phase than the fluorescent molecules. Small-angle X-ray scattering indicated that doxorubicin can reorganize anionic phospholipid dispersions into closely-packed multilamellar structures. Addition of the drug caused leakage of entrapped 6-carboxyfluorescein. Neither 2H-NMR on [2-2H]serine-labelled DOPS nor 31P-NMR revealed any significant effect of doxorubicin on headgroup conformation, but 2H-NMR on di[11,11-2H2]oleoyl-labelled phospholipids showed that the drug had a strong acyl chain-disordering effect on anionic phospholipids. 2H-NMR relaxation measurements indicated that the drug immobilized the headgroups and acyl chains of anionic phospholipids. The implications of these observations for the cellular activity of the drug are indicated.

Animals↗

Diets rich in eicosapentaenoic acid and gamma-linolenic acid affect phospholipid fatty acid composition and production of prostaglandins E1, E2 and E3 in turbot (Scophthalmus maximus), a species deficient in delta 5 fatty acid desaturase.

Duplicate groups of juvenile turbot, (Scophthalmus maximus), were fed diets containing either Marinol K (MO), a marine fish oil rich in eicosapentaenoic acid (EPA; 20:5, n-3) or borage oil (BO), rich in gamma-linolenic acid (GLA; 18:3, n-6), for a period of 12 weeks. Individual phospholipid fatty acid compositions from hearts of fish fed BO had significantly more 18:2, n-6, GLA, 20:2, n-6, dihomo-gamma-linolenic acid (DHGLA; 20:3, n-6) and total n-6 polyunsaturated fatty acids (PUFA), but significantly less arachidonic acid (AA; 20:4, n-6), compared to fish fed MO. In both phosphatidylcholine (PC) and phosphatidylethanolamine (PE) from heart, the DHGLA was increased by over 50-fold in fish fed BO while AA was reduced by over two-thirds, compared to fish fed MO. In brain, EPA was the major C20 PUFA, i.e. potential eicosanoid precursor in all phospholipids from fish fed MO, with the EPA level being twice that of AA in brain phosphatidylinositol (PI). DHGLA was the major C20 PUFA in all phospholipid classes from fish fed BO. In kidney and gill, EPA was the predominant C20 PUFA in all phospholipid classes, except PI, in fish fed MO. In kidney of fish fed BO, DHGLA was the major C20 PUFA in all phospholipid classes, except PE. In gill of fish fed BO, DHGLA was the major C20 PUFA in all phospholipid classes, including PI, where DHGLA was over 2.5-fold greater than AA. In homogenates of heart, kidney and gill from BO-fed fish the prostaglandin E1 (PGE1) concentration was significantly increased compared to MO-fed fish. In heart and kidney homogenates from fish fed MO the PGE3 concentration was significantly increased compared to fish fed BO. The ratio of PGE2/PGE1 was significantly reduced in brain, heart, kidney and gill homogenates from fish fed BO compared to those fed MO.

8,11,14-Eicosatrienoic Acid↗

Distribution of membrane phospholipids in the rabbit neural retina, optic nerve head and optic nerve.

Since diseases of the neural retina and optic nerve can result in alteration of biological membranes, this study determines similarities and differences in the membrane phospholipid content of the neural retina, optic nerve head, and optic nerve to serve as baseline data. Neural retina, optic nerve head, and optic nerve were dissected, isolated as 5 sets from 20 rabbits and frozen in liquid N2. Separate pooled-tissue extracts were prepared for each set of tissues and phosphorus-31 nuclear magnetic resonance (31P NMR) analyses performed. Ten phospholipids were quantified (respective neural retina, optic nerve head, and optic nerve mole % are given for the 5 major phospholipids detected): phosphatidylcholine (PC), 44.61, 27.67, 26.40; PC plasmalogen or alkylacyl PC (CPLIP); phosphatidylinositol (PI); sphingomyelin (SM); phosphatidylserine (PS), 12.63, 14.77, 15.09; phosphatidylethanolamine (PE), 21.21, 9.59, 8.69; PE plasmalogen (EPLAS), 11.07, 30.96, 33.93; an unidentified (unknown) phospholipid (U) at the chemical-shift value of 0.13 ppm; diphosphatidylglycerol (DPG); and phosphatidic acid (PA), 0.46, 2.92, 1.57. Significant differences between the various tissues were determined by the one-way analysis of variance, using a Scheffé range value of P < 0.05. The neural retina in all phospholipids detected except for the uncharacterized (unknown) phospholipid was significantly different from the optic nerve head tissue. The optic nerve head was significantly different from the optic nerve in PC, CPLIP, PE, EPLAS, U, DPG, and PA. The data provide a baseline for studies on pathologically changed neural retina, optic nerve head, and optic nerve.

Animals↗

Application of 31P MRS to the analysis of phospholipid changes in plasma of patients with acute leukemia.

The aim of the experiment was to evaluate the changes of phospholipid concentrations in patients (n=30) with acute leukemia compared with reference group of healthy volunteers (n=21). The analysis focused on the following phospholipids (PL) collected from plasma: phosphatidylcholine (PC), plasmalogen of phosphatidylcholine (CPLAS), lysophosphatidylcholine (LPC), sphingomyelin (SM), phosphatidylethanolamine (PE), and phosphatidylinositol (PI). Phospholipid extracts were obtained by Folch's method from 4 ml of plasma. 31P MR spectra were obtained on an AMX Bruker 300 MHz (7.05 T) spectrometer. Calculation of concentration based on integral intensity of the phospholipid relative to an internal concentration standard of MDPA. For healthy volunteers, the following values of phospholipid concentrations were obtained: (5.18+/-1.615) mmol/l for PC+CPLAS; (0.364+/-0.178) mmol/l for LPC; (1.211+/-0.411) mmol/l for SM; (0.343+/-0.124) mmol/l for PI+PE. PLs of patients were assayed at least twice: at the time of diagnosis and, when appropriate, at the time of complete remission from the disease (CR). At the time of diagnosis, the mean concentrations of studied compounds were: (1.602+/-0.716) mmol/l for PC+CPLAS; (0.041+/-0.048) mmol/l for LPC; (0.398+/-0.198) mmol/l for SM; (0.045+/-0.071) mmol/l for PI+PE. After attainment of complete remission (CR), the respective values were as follows: (4.094+/-1.886) mmol/l for PC+CPLAS; (0.295+/-0.139) mmol/l for LPC; (1.123+/-0.634) mmol/l for SM; (0.230+/-0.125) mmol/l for PI+PE. All concentrations found in patients at the time of diagnosis were significantly lower than in reference group and in those benefited from complete remission (CR). By contrast the differences in concentrations of phospholipids in plasma between patients with complete remission (CR) and healthy volunteers were no statistically significant.

Acute Disease↗

Cholesterol affects spectrin-phospholipid interactions in a manner different from changes resulting from alterations in membrane fluidity due to fatty acyl chain composition.

We previously showed that erythrocyte and brain spectrins bind phospholipid vesicles and monolayers prepared from phosphatidylethanolamine and phosphatidylserine and their mixtures with phosphatidylcholine (Review: A.F. Sikorski, B. Hanus-Lorenz, A. Jezierski, A. R. Dluzewski, Interaction of membrane skeletal proteins with membrane lipid domain, Acta Biochim. Polon. 47 (2000) 565). Here, we show how changes in the fluidity of the phospholipid monolayer affect spectrin-phospholipid interaction. The presence of up to 10%-20% cholesterol in the PE/PC monolayer facilitates the penetration of the monolayer by both types of spectrin. For monolayers constructed from mixtures of PI/PC and cholesterol, the effect of spectrins was characterised by the presence of two maxima (at 5 and 30% cholesterol) of surface pressure for erythroid spectrin, and a single maximum (at 20% cholesterol) for brain spectrin. The binding assay results indicated a small but easily detectable decrease in the affinity of erythrocyte spectrin for FAT-liposomes prepared from a PE/PC mixture containing cholesterol, and a 2- to 5-fold increase in maximal binding capacity (B(max)) depending on the cholesterol content. On the other hand, the results from experiments with a monolayer constructed from homogenous synthetic phospholipids indicated an increase in deltapi change with the increase in the fatty acyl chain length of the phospholipids used to prepare the monolayer. This was confirmed by the results of a pelleting experiment. Adding spectrins into the subphase of raft-like monolayers constructed from DOPC, SM and cholesterol (1/1/1) induced an increase in surface pressure. The deltapi change values were, however, much smaller than those observed in the case of a natural PE/PC (6/4) monolayer. An increased binding capacity for spectrins of liposomes prepared from a "raft-like" mixture of lipids could also be concluded from the pelleting assay. In conclusion, we suggest that the effect of membrane lipid fluidity on spectrin-phospholipid interactions is not simple but depends on how it is regulated, i.e., by cholesterol content or by the chemical structure of the membrane lipids.

Animals↗

Effects of lipid chain length on molecular interactions between paclitaxel and phospholipid within model biomembranes.

Molecular interactions between an anticancer drug, paclitaxel, and phosphatidylcholine (PC) of various chain lengths were investigated in the present work by the Langmuir film balance technique and differential scanning calorimetry (DSC). Both the lipid monolayer at the air-water interface and lipid bilayer vesicles (liposomes) were employed as model biological cell membranes. Measurement and analysis of the surface pressure versus molecular area curves of the mixed monolayers of phospholipids and paclitaxel under various molar ratio showed that phospholipids and paclitaxel formed a nonideal miscible system at the interface. Paclitaxel exerted an area-condensing effect on the lipid monolayer at small molecular surface areas and an area-expanding effect at large molecular areas, which could be explained by the intermolecular forces and geometric accommodation between the two components. Paclitaxel and phospholipids could form thermodynamically stable monolayer systems: the stability increased with the chain length in the order DMPC (C14:0)>DPPC (C16:0)>DSPC (C18:0). Investigation of paclitaxel penetration into the pure lipid monolayer showed that DMPC had a higher ability to incorporate paclitaxel and the critical surface pressure for paclitaxel penetration also increased with the chain length in the order DMPC>DPPC>DSPC. A similar trend was testified by DSC studies on vesicles of the mixed paclitaxel/phospholipids bilayer. Paclitaxel showed the greatest interaction with DMPC while little interaction could be measured in the paclitaxel/DSPC liposomes. Paclitaxel caused broadening of the main phase transition without significant change at the peak melting temperature of the phospholipid bilayers, which demonstrated that paclitaxel was localized in the outer hydrophobic cooperative zone of the bilayer. The interaction between paclitaxel and phospholipid was nonspecific and the dominant factor in this interaction was the van der Waals force or hydrophobic force. As the result of the lower net van der Waals interaction between hydrocarbon chains for the shorter acyl chains, paclitaxel interacted more readily with phospholipids of shorter chain length, which also increased the bilayer intermolecular spacing.

Calorimetry, Differential Scanning↗

Saturated phospholipids promote crystallization but slow down polymorphic transitions in triglyceride nanoparticles.

Some matrix materials proposed for the preparation of solid lipid nanoparticles (e.g. trilaurin) are difficult to crystallize after processing by melt-homogenization. In an attempt to overcome this difficulty, the effect of saturated long-chain phospholipids on the crystallization of nanoparticles based on trilaurin, trimyristin, tripalmitin and tristearin was studied. The phospholipids were used as emulsifiers in combination with sodium glycocholate. Saturated phospholipids increased the crystallization temperature of the triglyceride by several degrees compared to soybean phospholipids. The crystallization pattern was more complex in such systems due to solidification of the phospholipid chains prior to triglyceride crystallization. For most triglycerides, egg lecithin also induced crystallization at higher temperatures than natural soybean lecithin. With trilaurin dispersions, the effect of phospholipids can be utilized to induce crystallization at temperatures relevant for larger scale preparation. The polymorphic transitions of the triglycerides were slower in the presence of egg and saturated lecithin leading to a higher stability of the metastable alpha-form. These effects were particularly pronounced in tristearin systems where a predominant fraction of alpha-phase particles could be observed even after long-term cold storage in dispersions containing hydrogenated soybean lecithin or DPPC. The possibility to prepare triglyceride nanoparticles stable in specific modifications offers new opportunities to study effects of polymorphic form on colloidal stability, drug loading and release properties of such dispersions.

Calorimetry, Differential Scanning↗

The effects of cholesterol on magnetically aligned phospholipid bilayers: a solid-state NMR and EPR spectroscopy study.

This paper presents the first time that both solid-state NMR spectroscopy and EPR spectroscopy are used to study the effects of cholesterol on magnetically aligned phospholipid bilayers (bicelles). Solid-state deuterium NMR spectroscopy was carried out using both chain perdeuterated 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC-d(54)) and a partially deuterated beta-[2,2,3,4,4,6-(2)H(6)]cholesterol (cholesterol-d(6)). Also, EPR spectroscopy was carried out utilizing a 3 beta-doxyl-5 alpha-cholestane (cholestane) spin probe incorporated into magnetically aligned bilayers to provide a more complete picture about the ordering and dynamics of the phospholipid and cholesterol molecules in the bicelle membrane system. The results demonstrate that cholesterol was successfully incorporated into the phospholipid bilayers. The molecular order parameters extracted directly from the (2)H NMR spectra of both DMPC-d(54) and cholesterol-d(6) were compared to that from the EPR study of cholestane. The order parameters indicate that the sterol was motionally restricted, and that the DMPC had high order and low motion for the hydrocarbon segments close to the head groups of the phospholipids and less order and more rapid motion toward the terminal methyl groups. Both methods clearly indicate an overall increase in the degree of ordering of the molecules in the presence of cholesterol and a decrease in the degree of ordering at higher temperatures. However, EPR spectroscopy and (2)H NMR spectroscopy exhibit different degrees of sensitivity in detecting the phospholipid molecular motions in the membrane. Finally, cholesterol increases the minimum alignment temperature necessary to magnetically align the phospholipid bilayers.

Dimyristoylphosphatidylcholine↗

Effect of PA-MSHA vaccine on plasma phospholipids metabolic profiling and the ratio of Th2/Th1 cells within immune organ of mouse IgA nephropathy.

Phospholipids as a class of important constituents in the biomembranes have been paid increasing attention in many fields. IgA nephropathy is now generally known to be the most common form of primary glomerulonephritis in the world. However, phospholipids metabolism in IgA nephropathy was not clear. Until recently, there was no effective treatment available for patients with IgA nephropathy. In this paper, effect of PA-MSHA vaccine on plasma phospholipid metabolic profile of mouse IgA nephropathy was investigated using high performance liquid chromatography/mass spectrometry (HPLC/MS) and principal components analysis (PCA). Female Balb/c mice were divided into four groups: model group, control group, PA-MSHA treatment group and medicine control group (dipyridamole+common threewingnut root). The experimental IgA nephropathy model was established by the immunity combination method of oral BSA and injection of SEB. It was found that combination of LC/MS technology with PCA can be successfully applied to phospholipids profile analysis, clearly classify the model group and normal group, and PA-MSHA treatment group is closer to the normal control group than medicine control group. The result showed that Th(2)/Th(1) (=CD(4)(+)CD(30)(+)/CD(4)(+)CD(30)(-)) of the model group is 20.70+/-3.57, which is significantly higher than that of the control group (1.34+/-0.14) (P<0.001). The Th(2)/Th(1) ratio of the PA-MSHA treatment group and the medicine control group are lower than that of the model group (P<0.01). It is suggested that mouse IgA nephropathy has the phospholipids metabolic abnormality, PA-MSHA vaccine cannot only regulate the abnormal phospholipids metabolism mouse with the IgA nephropathy, but also correct the over unbalance of Th(2)/Th(1) proportion.

Animals↗