Effect of polyene phosphatidylcholine/ursodeoxycholic acid/ademetionine on pregnancy outcomes in intrahepatic cholestasis

BACKGROUND Intrahepatic cholestasis of pregnancy(ICP)is a liver disorder that occurs in pregnant women and can lead to a range of adverse pregnancy outcomes.The condition is typically marked by pruritus(itching)and elevated levels of liver enzymes and bile acids.The standard treatment for ICP has generally been ursodeoxycholic acid and ademetionine 1,4-butanedisulfonate,but the efficacy of this approach remains less than optimal.Recently,polyene phosphatidylcholine has emerged as a promising new therapeutic agent for ICP due to its potential hepatoprotective effects.AIM To evaluate the effect of polyene phosphatidylcholine/ursodeoxycholic acid/ademetionine 1,4-butanedisulfonate on bile acid levels,liver enzyme indices,and pregnancy outcomes in patients with ICP.METHODS From June 2020 to June 2021,600 patients with ICP who were diagnosed and treated at our hospital were recruited and assigned at a ratio of 1:1 via randomnumber table method to receive either ursodeoxycholic acid/ademetionine 1,4-butanedisulfonate(control group,n=300)or polyene phosphatidylcholine/ursodeoxycholic acid/ademetionine 1,4-butanedisulfonate(combined group,n=300).Outcome measures included bile acids levels,liver enzyme indices,and pregnancy outcomes.RESULTS Prior to treatment,no significant differences were observed between the two groups(P>0.05).Post-treatment,patients in both groups had significantly lower pruritus scores,but the triple-drug combination group had lower scores than the dual-drug combination group(P<0.05).The bile acid levels decreased significantly in both groups,but the decrease was more significant in the triple-drug group(P<0.05).The triple-drug group also exhibited a greater reduction in the levels of certain liver enzymes and a lower incidence of adverse pregnancy outcomes compared to the dual-drug group(P<0.05).CONCLUSION Polyene phosphatidylcholine/ursodeoxycholic acid/ademetionine 1,4-butanedisulfonate effectively relieves pruritus and reduces bile acid levels and liver enzyme indices in patients with ICP,providing a positive impact on pregnancy outcome and a high safety profile.Further clinical trials are required prior to clinical application.

Biliary phosphatidylcholine and lysophosphatidylcholine profiles in sclerosing cholangitis

AIM:To analyze phospholipid profiles in intrahepatic bile from patients with primary sclerosing cholangitis(PSC)and secondary sclerosing cholangitis(SSC).METHODS:Intrahepatic bile specimens collected via endoscopic retrograde cholangiography from 41 patients were analyzed.Fourteen of these patients were diagnosed with PSC,10 with SSC,11 with choledocholithiasis or no identifiable biliary disease,and 6 with cholangiocellular carcinoma(CCC).Bile acid,cholesterol,protein,and bilirubin contents as well as pancreas lipase activity in bile were determined by biochemical methods.Phosphatidylcholine(PC)and lysophosphatidylcholine(LPC)species were quantified using nanoelectrospray ionization tandem mass spectrometry.RESULTS:Bile from all the examined patient groups showed a remarkably similar PC and LPC species composition,with only minor statistical differences.Total biliary PC concentrations were highest in controls(8030±1843 mol/L)and lowest in patients with CCC(1969±981 mol/L)(P=0.005,controls vs SSC and CCC,respectively,P<0.05).LPC contents in bile were overall low(4.2%±1.8%).Biliary LPC/PC ratios and ratios of biliary PC to bilirubin,PC to cholesterol,PC to protein,and PC to bile acids showed no intergroup differences.CONCLUSION:PC and LPC profiles being similar in patients with or without sclerosing cholangitis,these phospholipids are likely not of major pathogenetic importance in this disease group.

Comparison Between the Influences of Phosphatidylcholine and Triton X-100 on the Protein Secondary Structures and Oxygen-evolving Activity of Photosystem Ⅱ

The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X_100 (TX_100) treatment were studied by using Fourier transform_infrared (FT_IR) spectroscopic technique and oxygen electrode. Phosphatidylcholine reconstitution showed no significant effect on the protein secondary structures of PSⅡ membrane but an increase of the rate of PSⅡ_mediated oxygen_evolution. The phosphatidylcholine lipids with different length of acyl chains displayed different capabilities to stimulate oxygen_evolution. In contrast, perturbation of the bilayer lipids by TX_100 resulted in obvious changes of the protein secondary structures within the PSⅡ membrane and in the loss of the PSⅡ_mediated oxygen_evolving activity. The results indicate the importance of membrane integrity in maintaining the stability of the photosynthetic membrane proteins.

Modeling, Simulation of a Simulated Moving Bed for Separation of Phosphatidylcholine from Soybean Phospholipids

A rate model, which considers axial dispersion, external mass transfer, intraparticle diffusion and nonlinear isotherms, and ports periodic switching is adopted to simulate the simulated moving bed （SMB） process. The effects of flow rate in Sections 2 and 3 and switching time on the operating performance parameters： purity, recovery, productivity and dcsorbent consumption are studied. A simulation approach is applied to simulate the operation and performance of the SMB. The model predicts the performance of the transient and cyclic steady state behavior to a reasonably good extent, and provides guidance operation condition of the SMB process.

LARGELY IMPROVED BLOOD COMPATIBILITY OF POLYURETHANE BY BLENDING WITH FLUORINATED PHOSPHATIDYLCHOLINE POLYURETHANE

The improvement of biocompatibility of polyurethanes was investigated.The results demonstrate that the blood compatibility of polyurethanes can be further improved by just simply mixing with the fluorinated phosphatidylcholine poly(carbonate urethane)s(FPCPCUs).The solution blending was done by mixing poly(ether urethane)(PEU)with FPCPCU in different compositions.An increased blood compatibility of the blend films was observed with the increase of FPCPCU content,and when FPCPCU content reached to 40 wt%(40F...

Moderating effect of synthesized docosahexaenoic acid-enriched phosphatidylcholine on production of Th1 and Th2 cytokine in lipopolysaccharide-induced inflammation

Objective: To evaluate effects of docosahexaenoic acid-enriched phosphatidylcholine(DHAPC) on cytokine production induced by lipopolysaccharide(LPS). Methods: The culture supernatants of splenocytes exposed to DHA-PC along with LPS were harvested to determine the production of Th 1(IFN-kines. Cytokines were m毭 and IL-2) and Th2 [IL-4, IL-5, IL-6 and IL-12/IL-23(p40)] cytoeasured using ELISA. Results: Co-administration of DHAPC with LPS resulted in significantly lower IL-2 expression compared to that observed with administration of only LPS(P<0.01). Treatment with DHA-PC and LPS significantly increased IL-5 expression(P<0.01). Moreover, co-administration of DHA-PC with LPS significantly decreased IL-6 and IL-12/IL-23(p40) expressions compared to that observed with administration of only LPS(P<0.01). Conclusions: Our results suggest that DHA-PC inhibits pro-inflammatory cytokines [IL-2, IL-6 and IL-12/IL-23(p40)] expression on induction of inflammation.

Separation of phosphatidylcholine from soybean phospholipids by simulated moving bed

A simulated moving bed (SMB), equipped with eight silica-gel columns, was used to separate phosphatidylcholine (PC) from soybean phospholipids. The effects of flow rate in Sections 2 (Q2) and 3 (Q3), switching time, feed flow rate and feed concentration on the operating performance parameters: purity, recovery, productivity and desorbent consumption were studied. Operating conditions leading to more than 90% purity in both outlet streams have been identified, together with those achieving optimal performance. Regions leading to complete separation are observed and explained theoretically. As the mass-transfer effect was not considered, the triangle theory only gives initial guesses for the optimal operating conditions.

HPLC Analysis of Egg Yolk Phosphatidylcholine by Evaporative Light Scattering Detector

Egg yolk phosphatidylcholine(EYPC) is being widely used in food and pharmaceutical industries nowadays owing to its surface activity,pharmaceutical usefulness,and so on.Common determination methods of phospholipids were based on the American Oil Chemists' Society(AOCS) Official Method Ja7b-91,in which n-hexane/2-propanol/acetate buffer was used as the mobile phase.In order to achieve desired results,gradient elu-tion or buffer solution was used,which made the detection process more complicated.Moreover,water or buffer solution could affect the silica gel column both on its lifespan and the separation efficiency significantly.In this study,different mobile phase and detector were used to simplify EYPC analyzing process instead of using water within the mobile phase.The optimized HPLC operating conditions are as follows:pure methanol as a mobile phase,flow rate of 1.0 ml·min-1,silica gel column(250 mm×4.6 mm,5 μm,Inertsil GLTM),column temperature 30 ℃ and low temperature evaporative light scattering detector(40 ℃,0.35 MPa) as used.Under this optimal condition,the linear relative coefficient of the standard curve is 0.998 and the recovery was in the range of 96.83%-101.58% with a relative standard deviation of 1.79%(n=6).

Formulation of self-nanoemulsifying drug delivery systems containing monoacyl phosphatidylcholine and Kolliphor^(■) RH40 using experimental design

The development of self-nanoemulsifying drug delivery systems(SNEDDS) to enhance the oral bioavailability of lipophilic drugs is usually based on traditional one-factor-at-a-time approaches. These approaches may be inadequate to analyse the effect of each excipient and their potential interactions on the emulsion droplet size formed when dispersing the SNEDDS in an aqueous environment. The current study investigates the emulsion droplet sizes formed from SNEDDS containing different levels of the natural surfactant monoacyl phosphatidylcholine to reduce the concentration of the synthetic surfactant polyoxyl 40 hydrogenated castor oil(Kolliphor ~? RH40). Monoacyl phosphatidylcholine was used in the form of Lipoid S LPC 80(LPC, containing approximately 80% monoacyl phosphatidylcholine, 13% phosphatidylcholine and 4% concomitant components). The investigated SNEDDS comprised of long-chain or medium-chain glycerides(40% to 75%), Kolliphor ~? RH40(5% to 55%), LPC(0 to 40%) and ethanol(0 to 10%). D-optimal design, multiple linear regression, and partial least square regression were used to screen different SNEDDS within the investigated excipient ranges and to analyse the effect of each excipient on the resulting droplet size of the dispersed SNEDDS measured by dynamic light scattering. All investigated formulations formed nano-emulsions with droplet sizes from about 20 to 200 nm. The use of mediumchain glycerides was more likely to result in smaller and more monodisperse droplet sizes compared to the use of long-chain glycerides. Kolliphor~? RH40 exhibited the most significant effect on reducing the emulsion droplet sizes. Increasing LPC concentration increased the emulsion droplet sizes, possibly because of the reduction of Kolliphor~? RH40 concentration. A higher concentration of ethanol resulted in an insignificant reduction of the emulsion droplet size. The study provides different ternary diagrams of SNEDDS containing LPC and Kolliphor ~? RH40 as a reference for formulation developers.

Effect of Sources and Concentrations of Soybean Phosphatidylcholine on Diluted Goat Semen Equilibrated at 4 ℃

The objective of the present study was to determine the proper sources and concentrations of soybean lecithin （phosphatidylcholine, PC） to be used as substitute for hen egg yolk in extender for preserving goat semen. Two sources of soybean lecithin （20% and 95% soybean phosphatidylcholine; PC20 and PC95） and three concentrations （1%, 2% and 3% v/v） of PC20 and PC95 supplemented in Tris-citric acid-fructose （TCF） extender were tested. The TCF extender supplemented with 20% hen egg yolk was used as a control. Fresh semen samples were collected from 3 goats by artificial vagina. Seminal plasma was removed by centrifugation and sperm pellets were pooled together and divided into 7 groups according to types of extender. The diluted semen samples were kept at 4 ℃ （equilibration）. The semen qualities including progressive motility, sperm viability, sperm plasma membrane integrity and tail abnormalities were evaluated before dilution and after 4 hrs equilibration. It was found that the progressive motility of equilibrated semen in egg yolk and PC20 extenders were higher than those in PC95 extender （P 〈 0.05）. Sperm viability was lower in 1% and 2% PC95 extender compared to other extenders （P 〈 0.05）. PC20 extender maintained the sperm membrane integrity and normal tail morphology at low temperature better than egg yolk and PC95 （P 〈 0.05）. It can be concluded that 20% soybean phosphatidylcholine supplemented in TCF extender at 1%-3% （v/v） is as effective as hen egg yolk to preserve goat semen during equilibration at 4 ℃ for 4 hrs .

Separate basolateral and apical phosphatidylcholine secretion routes in intestinally differentiated tumor cells

AIM:To investigate whether the secretion of phospha-tidylcholine(PC)in intestinal mucus occurs by apical secretion or via basolateral excretion and to determine its subsequent passage across the tight junctions to the apical mucus.METHODS:We addressed this question using the po-larized intestinally differentiated tumor cell line CaCo-2 grown on filters to confluence in Transwell culture chambers.The released PC and sphingomyelin(Sph)from apical and basolateral media were analyzed by mass spectrometry.RESULTS:The secreted PC species were identical in both compartments indicating the same intracellular origin of PC.However,PC secretion into the basolateral compart-ment was more effective,and the PC:Sph ratio in the ba-solateral compartment was signif icantly higher than that in the apical compartment(8.18 ± 1.84 vs 4.31 ± 1.22,P = 0.01).Both pathways were temperature sensitive and were unaltered in the presence of cyclosporine.CONCLUSION:The data demonstrate the PC secre-tion capacity of CaCo-2 cells and indicate two sepa-rated apical and basolateral release mechanisms.

Determination of Phosphatidylcholine Content in Zophobas morio L. by High Performance Liquid Chromatography

In this study, a high performance liquid chromatographic method was established to determine the content of phosphatidylcholine in Zophobas morio L. The samples were extracted with methyl alcohol, pttrified, analyzed by high performance liquid chromatography, and quantified with external standard method. The results showed a good linear correlation within the concentration range of 250 - 2 000 μg/ml, and the correlation coefficient was 0. 999 6. The standard recovery rate ranged from 97% to 109%, and the relative standard deviation was lower than 2.4%. Furthermore, phosphatidylcholine contents ~f Z. mor/o individuals at different growth stages were determined. According to the results, the variation pattern of phosphatidylcholine contents of Z. mor/o with individual weight was consistent with the binomial curve, and the correlation coefficient was R^2=0. 865 （P 〈0.01 ）. This study laid methodological foundation for the development of Z. mor/o functional products.

Effect of Mono-, Di-, and Trihydric Alcohols on Lipase-Catalyzed Alcoholysis of Phosphatidylcholine in Hexane

In this study, alcoholysis of phosphatidylcholine(PC) catalyzed by two lipases(Novozym 435 and Lipozyme RM IM) was performed in n-hexane to evaluate the effects of mono-, di-, and trihydric alcohols(ethanol, 1-butanol, 1,2-ethanediol,1,2-propanediol, and glycerol) on the alcoholysis reactions. In the reactions, as PC was converted into lysophosphatidylcholine(LPC),LPC was simultaneously alcoholyzed and converted into glycerophosphorylcholine(GPC). The alcohols affected the alcoholysis of both PC and LPC. When alcoholysis was catalyzed by Novozym 435, the reaction efficiencies of the selected alcohols in the alcoholysis of PC followed the order 1,2-propanediol ≈ ethylene glycol> 1-butanol > ethanol > glycerol, and the reaction efficiencies of these alcohols in the alcoholysis of LPC were 1,2-propanediol > ethylene glycol> 1-butanol ≈ glycerol > ethanol. For alcoholysis catalyzed by Lipozyme RM IM, the reaction efficiencies of alcohols in alcoholysis of PC followed the order ethylene glycol ≈1,2-propanediol > glycerol > ethanol > 1-butanol, and the reaction efficiencies in the alcoholysis of LPC were ethylene glycol >glycerol > 1,2-propanediol > ethanol > 1-butanol. In general, in the lipase-catalyzed alcoholysis of PC, reaction efficiencies with dihydric alcohols are higher than those with mono-and trihydric alcohols.

Rapid semi-quantification of triacylglycerols,phosphatidylcholines,and free fatty acids in the rice bran of one grain

We developed a microplate assay method for determining the contents of triacylglycerols（TAGs）, phosphatidylcholines（PCs）, and free fatty acids（FFAs） in the rice bran of one grain using enzymatic reactions. In this method, enzymes from commercially available kits were used. Optimum reaction conditions were established. It was found that Nonidet P-40 was the optimal among the three surfactants used（Triton X-100, Tween 40, and Nonidet P-40） when lipid was dissolved in a reaction solution. Using this method, it was possible to quantify TAGs, PCs, and FFAs in concentration ranges of 7–150, 5–70, and 8–200 mg L-（–1）, respectively. Furthermore, when the TAG contents in the rice bran were measured, the values closely corresponded to those obtained by extracting from large amounts of rice bran. However, sufficient data on the PC and FFA contents in rice bran are not available for valid comparisons. Although this method can accurately quantify the TAG contents in the rice bran of one grain, the accuracy of the PC and FFA contents has not been verified. Hence, future study is necessary.

Phosphatidylcholine deficiency increases ferroptosis susceptibility in the Caenorhabditis elegans germline

Ferroptosis,a regulated and iron-dependent form of cell death characterized by peroxidation of membrane phospholipids,has tremendous potential for the therapy of human diseases.The causal link between phospholipid homeostasis and ferroptosis is incompletely understood.Here,we reveal that spin-4,a previously identified regulator of the“B12-one-carbon cycle-phosphatidylcholine(PC)”pathway,sustains germline development and fertility by ensuring PC sufficiency in the nematode Caenorhabditis elegans.Mechanistically,SPIN-4 regulates lysosomal activity which is required for B12-associated PC synthesis.PC deficiency-induced sterility can be rescued by reducing the levels of polyunsaturated fatty acids,reactive oxygen species,and redox-active iron,which indicates that the sterility is mediated by germline ferroptosis.These results highlight the critical role of PC homeostasis in ferroptosis susceptibility and offer a new target for pharmacological approaches.

Dietary Lipid Intervention in the Prevention of Brain Aging

As people live longer,the burden of aging-related brain diseases,especially dementia,is increasing.Brain aging increases the risk of cognitive impairment,which manifests as a progressive loss of neuron function caused by the impairment of synaptic plasticity via disrupting lipid homeostasis.Therefore,supplemental dietary lipids have the potential to prevent brain aging.This review summarizes the important roles of dietary lipids in brain function from both structure and mechanism perspectives.Epidemiological and animal studies have provided evidence of the functions of polyunsaturated fatty acids(PUFAs)in brain health.The results of interventions indicate that phospholipids—including phosphatidylcholine,phosphatidylserine,and plasmalogen—are efficient in alleviating cognitive impairment during aging,with plasmalogen exhibiting higher efficacy than phosphatidylserine.Plasmalogen is a recognized nutrient used in clinical trials due to its special vinyl ether bonds and abundance in the postsynaptic membrane of neurons.Future research should determine the dose-dependent effects of plasmalogen in alleviating brain-aging diseases and should develop extraction and storage procedures for its clinical application.

Synthesis of biodegradable waterborne phosphatidylcholine polyurethanes for soft tissue engineering applications

To further improve the biocompatibility of polyurethanes,a new lysine 2-(2-aminoethoxy)ethanol phosphatidylcholine(LAPC)is synthesized to use as chain extender for preparing a series of phosphatidylcholine polyurethanes(PCPUs).Poly(e-caprolactone)(PCL)and poly(ethylene glycol)(PEG)are used as soft segments,and L-lysine ethyl ester diisocyanate(LDI),LAPC and L-lysine are used as hard segments.The obtained PCPUs exhibit appropriate mechanical properties with break elongation of 1000–1700%,tensile strength of 7–22 MPa,and relatively high elastic modulus of 11–18 MPa,which could admirably satisfy the requirement for soft tissue engineering scaffolds.The phosphatidylcholine structures can increase the hydrophilicity of PCPU surfaces,which effectively reduce protein adsorption and platelet adhesion while promoting the cell proliferation.In addition,the LAPC chain extender,PCPU films and ultimate degradation products of PCPUs are proved to be nontoxic in cytotoxicity test.More interestingly,the cytokine release test of macrophages manifests that both LAPC and PCPU degradation products could effectively improve the proliferation of macrophages and induce them into a wound-healing phenotype.Thus,the obtained PCPUs have greatly potential applications of soft tissue engineering scaffolds for tissue repair and wound healing.

Enhanced water solubility, antioxidant activity, and oral absorption of hesperetin by D-α-tocopheryl polyethylene glycol 1000 succinate and phosphatidylcholine

Hesperetin,an abundant bioactive component of citrus fruits,is poorly water-soluble,resulting in low oral bioavailability.We developed new formulations to improve the water solubility,antioxidant activity,and oral absorption of hesperetin.Two nano-based formulations were developed,namely hesperetin-TPGS(D-α-tocopheryl polyethylene glycol 1000 succinate)micelles and hesperetin-phosphatidylcholine(PC)complexes.These two formulations were prepared by a simple technique called solvent dispersion,using US Food and Drug Administration(FDA)-approved excipients for drugs.Differential scanning calorimetry(DSC)and dynamic light scattering(DLS)were used to characterize the formulations’physical properties.Cytotoxicity analysis,cellular antioxidant activity assay,and a pharmacokinetic study were performed to evaluate the biological properties of these two formulations.The final weight ratios of both hesperetin to TPGS and hesperetin to PC were 1:12 based on their water solubility,which increased to 21.5-and 20.7-fold,respectively.The hesperetin-TPGS micelles had a small particle size of 26.19 nm,whereas the hesperetin-PC complexes exhibited a larger particle size of 219.15 nm.In addition,the cellular antioxidant activity assay indicated that both hesperetin-TPGS micelles and hesperetin-PC complexes increased the antioxidant activity of hesperetin to 4.2-and 3.9-fold,respectively.Importantly,the in vivo oral absorption study on rats indicated that the micelles and complexes significantly increased the peak plasma concentration(Cmax)from 2.64μg/mL to 20.67 and 33.09μg/mL and also increased the area under the concentration–time curve of hesperetin after oral administration to 16.2-and 18.0-fold,respectively.The micelles and complexes increased the solubility and remarkably improved the in vitro antioxidant activity and in vivo oral absorption of hesperetin,indicating these formulations’potential applications in drugs and healthcare products.

Study on the Aggregation and Deaggregation Behaviors of Phosphatidylcholines

Phosphatidylcholines and their analogs, the functional building block of the membrane, are recently found to mediate multiple physiological processes and exhibit a broad range of desirable pharmacological effects, which involve hydrophobic lipophilic interactions (HLI) between the phospholipid and the cell membrane. The HLI behavior of phosphatidylcholines (Ln) and their analogues 1, 2-diacyI-sn-glycerol-3-phosphoric add bro-moethyl ester (Pn), have been investigated in MeOH-H2O binary systems of different volume fractions (designated as Φ) of the organic component, by employing a-nephthylethyl lauryl ether (Np-12) as fluorescent probe. A very interesting observation is that the Ln possesses double character, i.e., it behaves both as an aggregator and as a deaggregator. The effects of the structure and the environment on the coaggregation and deag-gregation are also discussed.

Reduced phosphatidylcholine synthesis suppresses the embryonic lethality of seipin deficiency

Seipin plays a vital role in lipid droplet homeostasis,and its deficiency causes congenital generalized lipodystrophy type II in humans.It is not known whether the physiological defects are all caused by cellular lipid droplet defects.Loss-of-function mutation of seip-1,the Caenorhabditis elegans seipin ortholog,causes embryonic lethality and lipid droplet abnormality.We uncover nhr-114 and spin-4 as two suppressors of seip-1 embryonic lethality.Mechanistically,nhr-114 and spin-4 act in the“B12-one-carbon cycle-phosphatidylcholine(PC)”axis,and reducing PC synthesis suppresses the embryonic lethality of seip-1 mutants.Conversely,PC deficiency enhances the lipid droplet abnormality of seip-1 mutants.The suppression of seip-1 embryonic lethality by PC reduction requires polyunsaturated fatty acid.In addition,the suppression is enhanced by the knockdown of phospholipid scramblase epg-3.Therefore,seipin and PC exhibit opposite actions in embryogenesis,while they function similarly in lipid droplet homeostasis.Our results demonstrate that seipin-mediated embryogenesis is independent of lipid droplet homeostasis.