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.

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.

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.

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.

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.

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.

LRH-1 senses signaling from phosphatidylcholine to regulate the expansion growth of digestive organs via synergy with Wnt/β-catenin signaling in zebrafish

Liver receptor homolog-1(LRH-1)is an orphan nuclear receptor that is critical for the growth and proliferation of cancer cells and other biological processes,including lipid transportation and metabolism,sexual determination and steroidogenesis.However,because homozygous lrh-1mice die in utero,the regulatory mechanisms involved in embryonic development mediated by this receptor are poorly understood.In the present study,we performed transcription activator-like effector nuclease(TALEN)-mediated loss-of-function assays,taking advantage of zebrafish external fertilization,to investigate the function of lrh-1.The digestive organs were affected by lrh-1 depletion as a result of cellcycle arrest(at the checkpoint of G1 to S phase),but not cell apoptosis.Biochemical analysis revealed that LRH-1 augments the transcriptional activity of b-catenin 1 and 2 via physical interactions.Screening the specific ligand(s)sensed by LRH-1 during organogenesis revealed that phosphatidylcholine(PC),a potential ligand,is the upstream target of LRH-1 during endoderm development.These data provide evidence for the crosstalk between the PC/LRH-1 and Wnt/β-catenin signaling pathways during the expansion growth of endoderm organs.

Transmembrane Ca^(2+) gradient-mediated phosphatidylcholine modulating sarcoplasmic reticulum Ca^(2+)-ATPase

The sarcoplasmic reticulum (SR) Ca2+-ATPase was purified and reconstituted into the sealed phospholipids vesicles with or without transmembrane Ca2+ gradient. The role ofphospholipids, especially phosphatidylcholine(PC), in the modulation of Ca2+-ATPase by transmembrane Ca2+ gradient was investigated. The results are as follows, (i) Incubated with phospholiplds, the enzyme activity of the delipidated Ca2+-ATPase is inhibited by Ca2+ and the highest inhibition is observed in the presence of PC. (ii) When there exists a transmembrane Ca2+ gradient (higher Ca2+ concentration inside vesicles, 1 000μmol/L:50μmol/L, similar to the physiological condition), the inhibition of Ca2+-ATPase by transmembrane Ca2+ gradient can be only observed in the vesicles containing PC:PE, but not in those containing PS:PE or PG:PE. The highest inhibition is obtained at a 50.50 molar ratio of PC:PE. (iii) By comparing the effects of PC differing in acyl chains, higher inhibition of Ca2+-ATPase is observed in vesicles

Effect of Phosphatidylcholine on the Steady State Fluorescence of Chlorophyll in Photosystem Ⅱ Particles

Phosphatidylcholine (PC) accounts for less than 1% of the total lipids in plant photosystem II (PSII) particles. In this experiment, PSII particles were reconstituted with PC to construct PSII\|PC vesicles. The effect of PC on the steady state fluorescence of chlorophyll (Chl) in PSII particles was studied. The results show that PC significantly affected the fluorescence intensity, but did not obviously affect the fluorescence emission band peak position. PC also did not obviously affect the absorbance at 436?nm or the amide I band peak position in FT\|IR spectroscopy of PSII particles. The results suggest that PC may affect the light energy transfer from the antenna chlorophyll molecules to the reaction center chlorophyll molecule (P680).

BIOMIMETIC SURFACE PREPARATION OF INERT POLYMER FILMS VIA GRAFTING LONG MONOALKYL CHAIN PHOSPHATIDYLCHOLINE

To explore construction of novel mimicking biomembrane on biomaterials surfaces, a new polymerizable phosphatidylcholine containing a long monoalkyl chain ended with acryl group （AASOPC） was designed and synthesized, which was easily derived from the terminal amino group of 9-（2-amino-ethylcarbamoyl）-nonyl-l-phosphatidyl-choline （ASOPC） reacting with acryloyl chloride. The obtained AASOPC was grafted on poly（ethylene terephthalate） （PET） via surface-initiated atom-transfer radical polymerization （SI-ATRP） to form mimicking biomembrane. These modified surface structures of PET were investigated using water contact angle （WAC）, X-ray photoelectron spectroscopy （XPS） and atomic force microscopy （AFM）. The results indicated that the new mimicking phosphatidylcholine biomembrane could be prepared on inert polymer surfaces by using the acryloyl phosphatidylcholine （AASOPC） via surface-initiated atom transfer radical polymerization （SI-ATRP）.

Enhanced Ionization of Phosphatidylcholines during MALDI Mass Spectrometry Using DCTB as Matrix

Phosphatidylcholines （PCs） are the most abundant phospholipids constituents of the cellular membrane, which exhibit a variety of biological functions. The distinct critical roles in cellular function make their analysis quite de- manding. Although MALDI-MS is one of the most powerful tools for biomolecules identification, it is still limited to phospholipids studies. The ionization of phosphatidylcholines is insufficient, and signals of PCs are frequently confused and suppressed by matrix clusters occurring in the same mass range. As an alternative matrix, T-2-（3-（4-t-butyl-phenyl）-2-methyl-2-propenylidene） malononitrile （DCTB） was introduced to overcome these problems in our study. Specifically, the signal intensity of phosphatidylcholines from soybean was enhanced more than several ten-folds using DCTB during positive ion MALDI-TOFMS. Peak overlaps caused by the wide distri- butions of series fatty acid residues from phospholipids were separated by introducing cesium cation. The occurred mass shift of 131.90 Da between [M＋Cs]＋ and [M＋H]~ （＂M＂ represents the molecular weight of the corre- sponding neutral hosphatidylcholine） was approved to be helpful in the assignments of ambiguous peaks of PCs from soybean. For real sample analysis, employing cesium chloride as an auxiliary reagent successfully facilitated the profiling and characterizing of PCs extracted from mouse lung and egg yolk followed by analysis with MALDI-TOFMS using DCTB as matrix.

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.

Kinetic studies on the aggregation behavior of phosphatidylcholines and their analogs

Phosphatidylcholines (Ln's) and their neutral analogs 1,2-diacyl-sn-glycerol-3-phosphoric acid bromoethyl esters (Pn's) have been found to exhibit different aggregation behaviors brought about by hydrophobic-lipophilic interactions (HLI) by means of kinetic probe in MeOH-H2O binary aquiorganic systems. The effects of the structure and the environment on the interesting aggregation of the amphiphilic molecules are discussed.

Changes of adipocytokine expression after diabetic rats received sitagliptin and the molecular mechanism

Objective:To study the effect of sitagliptin on adipocytokines expression in diabetic rats and its molecular mechanism.Methods:Male SD rats were chosen and randomly divided into NC group,T2DM group,SP group and SP+LY group.NC group received conventional breeding,T2DM group,SP group and SP+LY group received intraperitoneal injection of streptozotocin after 12 weeks of high-fat diet to establish diabetes animal model,SP group received sitagliptin intervention and SP+LY group received sitagliptin combined with PI3 K inhibitor LY294002 intervention.6 weeks after the intervention,serum was collected to determine the levels of biochemical indexes and adipocytokines,and visceral adipose tissue was collected to determine expression levels of adipocytokines.Results:Serum TC,TG,LDL-C,FBG,FINS,Leptin and Chemerin levels as well as HOMA-IR of T2DM group were higher than those of NC group,and HDL-C,Adiponectin and Omentin-1 levels were significantly lower than those of NC group; serum TC,TG,LDL-C,FBG,FINS,Leptin and Chemerin levels as well as HOMA-IR of SP group were lower than those of T2DM group,and HDL-C,Adiponectin and Omentin-1 levels were significantly higher than those of T2DM group; Leptin and Chemerin levels in serum and visceral adipose tissue of SP+LY group were higher than those of SP group while Adiponectin and Omentin-1 levels were significantly lower than those of SP group.Conclusion:Sitagliptin can regulate the expression of adipocytokines in adipose tissue of diabetic rats through PI3K-AKT pathway.

Visualization of sphingolipids and phospholipids in the fundic gland mucosa of human stomach using imaging mass spectrometry

AIM:To analyze the lipid distribution in gastric mucosae.METHODS:Imaging mass spectrometry(MS)is a useful tool to survey the distribution of biomolecules in surgical specimens.Here we used the imaging MS apparatus named i MScope to identify the dominant molecules present in the human gastric mucosa near the fundic glands.Five gastric specimens were subjected to iM Scope analysis.These specimens were also analyzed by immunohistochemistry using MUC5 AC,H(+)-K(+)-ATPaseβ Claudin18 antibodies.RESULTS:Three major molecules with m/z 725.5,780.5,and 782.5 detected in the gastric mucosa were identified as sphingomyelin(SM)(d18:1/16:0),phosphatidylcholine(PC)(16:0/18:2),and PC(16:0/18:1),respectively,through MS/MS analyses.Using immunohistological staining,SM(d18:1/16:0)signals were mainly colocalized with the foveolar epithelium marker MUC5 AC.In contrast,PC(16:0/18:2)signals were observed in the region testing positive for the fundic gland marker H(+)-K(+)-ATPaseβ.PC(16:0/18:1)signals were uniformly distributed throughout the mucosa.CONCLUSION:Our basic data will contribute to the studies of lipid species in physical and pathological conditions of the human stomach.