Carboxyl Ester Lipase Protects Against Metabolic Dysfunction-Associated Steatohepatitis by Binding to Fatty Acid Synthase

Carboxyl ester lipase(CEL),a pivotal enzyme involved in lipid metabolism,is recurrently mutated in obese mice.Here,we aimed to elucidate the functional significance,molecular mechanism,and therapeutic potential of CEL in metabolic dysfunction-associated steatohepatitis(MASH).Hepatocyte-specific carboxyl ester lipase gene(Cel)knockout(Cel^(DHEP))and wildtype(WT)littermates were fed with cholinedeficient high-fat diet(CD-HFD)for 16 weeks,or methionine-and choline-deficient diet(MCD)for three weeks to induce MASH.Liquid chromatography–mass spectrometry and co-immunoprecipitation were employed to identify the downstream targets of CEL.CD-HFD/MCD-fed WT mice received intravenous injections of CEL-adeno-associated viral,serotype 8(AAV8)to induce specific overexpression of CEL in the liver.We observed a decrease in CEL protein levels in MASH induced by CD-HFD or MCD in mice.Cel^(DHEP) mice fed with CD-HFD or MCD exhibited pronounced hepatic steatosis,inflammation,lipid peroxidation,and liver injury compared to WT littermates,accompanied by increased hepatic nuclear factor kappa-light-chain-enhancer of activated B cell(NF-jB)activation.Consistently,Cel knockdown in mouse primary hepatocytes and AML12 cells aggravated lipid accumulation and inflammation,whereas CEL overexpression exerted the opposite effect.Mechanistically,CEL directly bound to fatty acid synthase(FASN),resulting in reduced FASN SUMOylation,which in turn promoted FASN degradation through the proteasome pathway.Furthermore,inhibition of FASN ameliorated hepatocyte lipid accumulation and inflammation induced by Cel knockdown in vivo and in vitro.Hepatocyte-specific CEL overexpression using AAV8-Cel significantly mitigated steatohepatitis in mice fed with CD-HFD or MCD.CEL protects against steatohepatitis development by directly interacting with FASN and suppressing its expression for de novo lipogenesis.CEL overexpression confers a therapeutic benefit in steatohepatitis.

Enzymatic synthesis, antioxidant ability and oil-water distribution coefficient of troxerutin fatty acid esters

Troxerutin fatty acid esters were prepared using troxerutin and fatty acid vinyl esters as substrates in pyridine through enzymatic route. The structures of as-prepared compounds were identified by FT-TR, NMR, and ESI-HRMS. Using alkaline protease(≥30 mg/mL) as enzyme, maximum yields reached 58% at 3:1(vinyl hexanoate to troxerutin) in pyridine(water content ≤1%). The yields gradually declined as chain length of acyl donors rose. The antioxidation abilities of the as-obtained compounds were confirmed by both DPPH free radical scavenging and potassium ferricyanide reduction methods. The antioxidation ability of troxerutin fatty acid esters was found lower than that of troxerutin. However, the logP values of troxerutin fatty acid esters varied from 0.15 to 1.94, suggesting that troxerutin fatty acid esters had better lipophilicity than troxerutin(logP =-2.12) when compared to their oil-water distribution coefficients. Overall, these findings look promising as reference for further development of future troxerutin.

Synthesis, Crystal Structure, and Herbicidal Activity of(3R,4R)-4,7,7-Trimethyl-6-oxabicyclo[3.2.1]octane-3,4-diol Mono-fatty Acid Esters

Four mono-fatty acid esters(Ia～d) were synthesized via the esterification of(3R,4R)-4,7,7-trimethyl-6-oxabicyclo[3.2.1]octane-3,4-diol(I) with fatty acid chlorides(CH_3(CH_2)_nCOCl, n = 0, 2, 4, 6) and structurally characterized by means of H RMS, IR, ~1 H-NMR, ^(13)C-NMR and X-ray diffraction. Compounds Ia～d all belong to monoclinic system, P2_1/c space group. Intermolecular O(2)–H(2)···O(1) hydrogen bonds, intramolecular O(2)–H(2)···O(3) hydrogen bonds and van der Waals' interaction between fatty acid ester groups link each of the mono-fatty acid ester molecules into a bilayer structure similar to liposome with the exposed hydrophobic moiety and the sandwiched lipophilic moiety. Especially, compounds Ia～d could be dissoluble or scattered in aqueous solution and showed hydrophilic/lipophilic property-dependent herbicidal activity against the dicotyledon plant rape(Brassica campestris) and the monocotyledon plant barnyard grass(Echinochloa crus galli). At the concentration of 10 mmol·L^(-1), the inhibition rates of compounds Ia～d against the root growth of rape are 31.9, 90.8, 99.5 and 100%, respectively and the inhibition rates against the shoot elongation of barnyard grass are 19.3, 50.0, 80.2 and 100%, respectively.

Epoxidation of Unsaturated Fatty Acid Methyl Esters in the Presence of SO_3H-functional Brφnsted Acidic Ionic Liquid as Catalyst

The epoxidation of unsaturated fatty acid methyl esters(FAMEs)by peroxyacetic acid generated in situ from hydrogen peroxide and acetic acid was studied in the presence of SO3H-functional Brnsted acidic ionic liquid (IL)[C3SO3HMIM][HSO4]as catalyst.The effects of hydrogen peroxide/ethylenic unsaturation ratio,acetic acid concentration,IL concentration,recycling of the IL catalyst,and temperature on the conversion to oxirane were studied.The kinetics and thermodynamics of unsaturated FAMEs epoxidation and the kinetics of oxirane cleavage of the epoxidized FAMEs by acetic acid were also studied.The conversion of ethylenic unsaturation group to oxirane, the reaction rate of the conversion to oxirane,and the rate of hydrolysis(oxirane cleavage)were higher by using the IL catalyst.

Studies on Production of Biodiesel by Esterification of Fatty Acids by a Lipase Preparation from Candida sp. 99-125

A self-made lipase preparation from Candida sp. 99-125 was used for the production of biodiesel through enzymatic esterification of fatty acids. The crude lipase powder and fermentation broth were immobilized on a cheap fiber cloth carrier. The conditions of lipase-catalyzed esterification between long-chain fatty acids and methanol in a solvent system were investigated in detail, including the temperature, pH value, substrate concentration, solvent, absorbent agent, enzyme dosage and purity, immobilization method, the mode of addition of substrate. The results show that reaction temperature, pH of lipase micro-environment, substrate concentration, enzyme dosage and purity affect the esterification strongly. Several new methods and enzymatic procedures for improving the enzymatic reaction involving the process cost are also discussed, such as fossil diesel fuel as reaction solvent, immobilization method, multi-step gradient addition of methanol. The esterification degree of 92.8% was obtained with oleic acid and methanol under the optimal reaction condition after 12.5 h reaction time. The half-life of the immobilized lipase preparation from crude free lipase powder for esterification was 15 days.

Inhibitory effect of schisandrin B on free fatty acid-induced steatosis in L-02 cells

AIM:To investigate the effects of schisandrin B (Sch B) on free fatty acid (FFA)-induced steatosis in L-02 cells.METHODS:Cellular steatosis was induced by incubating L-02 cells with a FFA mixture (oleate and palmitate at the ratio of 2:1) for 24 h.Cytotoxicity and apoptosis were evaluated by 3-(4,5-dmethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and Annexin V/propidium iodide staining,respectively.Cellular total lipid was determined using a photocolorimetric method after Nile red staining,and triglyceride content was measured using an enzymatic kit.To study the effects of Sch B on steatosis,L-02 cells were treated with Sch B (1-100 μmol/L) in the absence or presence of 1 mmol/L FFA for 24 h,and cellular total lipid and triglyceride levels were measured.To explore the mechanisms of action of Sch B in the steatotic L-02 cells,mRNA levels of several regulators of hepatic lipid metabolism including adipose differentiation related protein (ADRP),sterol regulatory element binding protein 1 (SREBP-1),peroxisome proliferator-activated receptor (PPAR)-α and PPAR-γ were measured by quantitative real-time polymerase chain reaction (PCR),and protein levels of ADRP and SREBP-1 were measured by immunoblotting.RESULTS:Treatment with 1 mmol/L FFA for 24 h induced intracellular lipid accumulation in L-02 cells comparable to that in human steatotic livers without causing apparent apoptosis and cytotoxicity.Sch B mitigated cellular total lipid and triglyceride accumulations in the steatotic L-02 cells in a dose-dependent manner.Quantitative real-time PCR and Western blot analyses revealed that treatment of L-02 cells with 100 μmol/L Sch B reverted the FFA-stimulated up-regulation of ADRP and SREBP-1.CONCLUSION:Sch B inhibits FFA-induced steatosis in L-02 cells by,at least in part,reversing the up-regulation of ADRP and SREBP-1.

Boosted Biodegradability and Tribological Properties of Mineral Base Oil by Methyl Diethanolamine Fatty Acid Esters

Methyl diethanolamine fatty acid esters, viz. methyl diethanolamine octanoate and methyl diethanolamine oleate,were prepared. Their impacts on the biodegradability and tribological properties of mineral base oil 400 SN were evaluated by a tester for fast evaluating the biodegradability of lubricants and by a four-ball tester, respectively. The results showed that methyl diethanolamine octanoate and methyl diethanolamine oleate both could markedly promote the biodegradation of the oil and improved its tribological properties. The improvement of biodegradability was attributed to the enhanced growth and quantity of microbes by methyl diethanolamine fatty acid esters. The worn surfaces were analyzed by a scanning electron microscope(SEM) equipped with an energy dispersive spectrometer(EDS) and an X-ray photoelectron spectroscope(XPS). The results indicated that the enhancement of friction-reducing and anti-wear properties of the mineral oil was attributed to the formation of complicated boundary lubrication films composed of species such as Fe_2O_3, Fe_3O_4 and organic nitrogen-containing compounds with a structure of –C-N-or R-NH_2.

Synthesis and Primary Research on Antitumor Activity of Three New Panaxadiol Fatty Acid Esters

For making use of Ginseng resources that exhibit an antitumor activity and for finding new anticancer drugs, three new fatty acid ester compounds： 3/β-acetoxy panaxadiol （ Ⅰ ）, 3β-palmitic acid aceloxy panaxadiol （ Ⅱ ） , and 3β-octadecanoic acid aceloxy panaxadiol（ Ⅰ , Ⅱ, and m） were synthesized with panaxadiol, diacetyl oxide, palmityl chloride and stearyl chloride, and their structures were determined via MS, ^13C NMR, IR, TLC, and so on. The molar yields of the three compounds are 75.14%, 79. 08%, and 72. 57%, respectively. Meanwhile, the antitumor activity of the three new panaxadiol fatty acid ester derivatives and panaxadiol was compared by using the method of MTT. Tumor cell used was Vero cell line. Positive control was 5-FU, blank was an RPMI1640 culture medium, negative control was an RPMI1640 culture medium and the solvent for drugs to be tested. Compound Ⅰ has the strongest antitumor activity followed by panaxadiol; compounds Ⅱ and Ⅲ have similar and weakest antitumor activities. Furthermore, the antitumor activities of the panaxadiol fatty acid ester derivatives show positive correlation with the concentration of the test group, but show no relationship with the molecular weight of fatty acid. The methods that are used to synthesize the three compounds with high yields and strong antitumor activities are simple and show a great potential for meeting the needs of industrial manufacture of these drugs.

Preparation of Highly Concentrated Silver Nanoparticles in Reverse Micelles of Sucrose Fatty Acid Esters through Solid-Liquid Extraction Method

Silver nanoparticles were synthesized in reverse micelles consisting of sucrose fatty acid esters by dissolving reactant powder in the water pool of reverse micelles through the solid-liquid extraction. Silver nanoparticles having various sizes and shapes were obtained at high concentration. The size of silver nanoparticles was controlled by reaction temperature. Moreover, the size of silver nanoparticles was dependent upon the average esterification degree of sucrose fatty acid esters forming reverse micelles. The wavelength in the peaks, which corresponded upon the localized surface plasmon resonance of resultant silver nanoparticles, was correlated with their sizes.

Investigation of <i>β</i>-Cryptoxanthin Fatty Acid Ester Compositions in Citrus Fruits Cultivated in Japan

In this study, a catalog of β-cryptoxanthin (βCX) and its fatty acid esters (βCXFAs) in citrus fruits were constructed. The peel and pulp of citrus fruits from nine Japanese cultivars and one foreign cultivar were surveyed by supercritical fluid chromatography/triple quadrupole mass spectrometry (SFC/QqQMS). In each citrus sample, the major components were βCX and its laurate ester, myristate ester, palmitate ester, and oleate ester. However, the composition ratio of βCXFA varied with the citrus breed and between the pulp and the peel of the fruit in most cultivars. The SFC/QqQMS system could be used to obtain new information about βCXs and carry out further discussion on the feature and tendency of each citrus variety.

Effects of Yttrium Doping on the Performance of Ru-Based Catalysts for Hydrogenation of Fatty Acid Methyl Ester

The highly dispersed supported ruthenium-yttrium (Ru-Y) bimetallic catalysts were prepared by impregnation method and their catalytic performance for hydrogenation of ester was fully investigated. The catalyst was characterized by X-ray diffraction and field emission scanning electron microscopy. The results show that the average particle diameter of the bimetallic crystallites was less than 10 nm. The effects of the reaction temperature, the hydrogen pressure, the amount of catalyst and the proportion of yttrium in catalyst on the hydrogenation of ester were studied. The experimental results show that the introduction of yttrium not only changed the chemical and textural properties of ruthenium-based catalyst but also controlled the formation of Ru-Y alloy. The Ru-Y catalyst (Ru-2%Y/TiO2) exhibited high catalytic activity and good selectivity towards the higher alcohols. Under optimal reaction conditions of 240°C and 5 MPa hydrogen pressure, the conversion of palm oil esters was above 93.4% while the selectivity towards alcohol was above 99.0%.

Optimization of Wax Esters Production from Palm Fatty Acid Distillate and Oleyl Alcohol over Amberlyst 15

Wax esters were derived from long chain fatty acids and long chain alcohols with chain length of 12 carbons or more. These compounds have many potential applications in cosmetics, pharmaceuticals and food industries. The present work focuses on the synthesis of wax esters using palm fatty acid distillate and oleyl alcohol catalyzed by Amberlyst 15. Response surface methodology （RSM） based on a five-level, three-variable central composite design （CCD） was used to evaluate the interactive effects of synthesis, of amount of Amberlyst 15 catalyst （21.6-38.4% w/w）, reaction time （18-102 min） and molar ratio （palm fatty acid distillate to oleyl alcohol, 1：1.16-1：2.84） on the percentage conversion of palm fatty acid distillate. The optimum conditions derived via RSM were： amount of catalyst 33% w/w, reaction time 95 minute and palm fatty acid to oleyl alcohol molar ratio 1：2.7. The actual experimental conversion was 81.52% under optimum condition, which compared well to the maximum predicted value of 80.50%. Analysis of the yield showed that at optimum condition, 80.54% wax esters were produced.

Emulsification Properties of Lactose Fatty Acid Esters

Sugar fatty acid esters (SFAE) are a class of synthetic emulsifiers used in the food, pharmaceutical, and personal care industries. The influence of the fatty acid chain length on the emulsification properties of lactose fatty acid esters (LFAE) including lactose monooctanoate (LMO), lactose monodecanoate (LMD), lactose monolaurate (LML) and lactose monomyristate (LMM) was investigated in this study. The stability of the emulsions as well as the oil droplet size distribution in 20% soybean oil-in-water emulsions was measured at 0.1%, 0.25% and 0.5% of LFAE concentrations. In order of LFAE with the strongest emulsion stabilization characteristics were LML, LMD, LMO and LMM. Oil droplet distributions resulted in the same trend, with LML and LMD maintaining the smallest droplet sizes and thus the most stable emulsion. The hydrophilic-lipophilic balance (HLB) and critical micelle concentrations were determined for each LFAE. An increase in HLB value was seen with an increased CMC value for each LFAE, showing the strength of the linear relationship between these two measured values. Additionally, there was a decrease in HLB and CMC values with a decrease in the fatty acid chain length of each LFAE. This research showed that LML and LMD formed more stable emulsions, even with HLB and CMC values higher than those of LMM suggesting HLB and CMC values alone do not predict emulsifier effectiveness.

Ternary Liquid-Liquid Equilibrium for Systems of Fatty Acid Methyl Ester(Methyl Palmitate/Methyl Stearate)+Ethanol+Glycerol at Atmospheric Pressure

Liquid-liquid equilibrium data of two ternary systems methyl palmitate+ethanol+glycerol and methyl stearate+ethanol+glycerol at(318.2 and 333.2)K and atmospheric pressure were measured. The values of distribution coefficient and selectivity were calculated, which indicates that glycerol can be separated from fatty acid ester by using ethanol as an extraction solvent. The consistency of the isothermal tie-line data were checked by the Othmer-Tobias equation. The correlation coefficients R2 are higher than 0.993,9 for all the fitted curves. The NRTL activity coefficient model was applied to the correlation of the measured tie-line data. The root mean square deviation(RMSD)values are less than 0.007 for all the systems, which shows a good predictive capability of this model for such systems.

Artificial Triterpenoid Fatty Acid Ester Isolated From the Leaves of Phytolacca icosandra L

The methanol extract form the leaves of Phytolacca icosandra L.,afforded the unprecedented artificial triterpenoid fatty acid ester 1 derived from the new natural triterpenoid phytolaccagenic acid 3-O-myristate(1a),along with the three known triterpenoids serjanic,acinosolic and phytolaccagenic acid(2-4).Their structures were stablished by HR-EI-MS,1D and 2D NMR techniques.The possible mechanistic formation of 1 is proposed,and the in vitro toxicity of all compounds was assessed using the brine shrimp lethality assay(BSLA).

Reduced lysosomal acid lipase activity: A new marker of liver disease severity across the clinical continuum of non-alcoholic fatty liver disease?

Lysosomal acid lipase(LAL)plays a key role in intracellular lipid metabolism.Reduced LAL activity promotes increased multi-organ lysosomal cholesterol ester storage,as observed in two recessive autosomal genetic diseases,Wolman disease and Cholesterol ester storage disease.Severe liver steatosis and accelerated liver fibrosis are common features in patients with genetic LAL deficiency.By contrast,few reliable data are available on the modulation of LAL activity in vivo and on the epigenetic and metabolic factors capable of regulating its activity in subjects without homozygous mutations of the Lipase A gene.In the last few years,a less severe and non-genetic reduction of LAL activity was reported in children and adults with non-alcoholic fatty liver disease(NAFLD),suggesting a possible role of LAL reduction in the pathogenesis and progression of the disease.Patients with NAFLD show a significant,progressive reduction of LAL activity from simple steatosis to non-alcoholic steatohepatitis and cryptogenic cirrhosis.Among cirrhosis of different etiologies,those with cryptogenic cirrhosis show the most significant reductions of LAL activity.These findings suggest that the modulation of LAL activity may become a possible new therapeutic target for patients with more advanced forms of NAFLD.Moreover,the measurement of LAL activity may represent a possible new marker of disease severity in this clinical setting.

Lipids and fatty acids in Calanus sinicus during over-summering in the southern Yellow Sea

Over-summering is a crucial period for Calanus sinicus in the southern Yellow Sea,where it is a key member of the zooplankton community.Lipids play an important role in copepod diapause,which is part of their over-summering strategy.We investigated how different fatty acids and lipid classes,including wax esters,changed during over-summering of C.sinicus during three cruises in June and August 2011 and November 2010,corresponding to the pre-,during and post-diapause periods,respectively.Large amounts of lipids were accumulated,mainly wax esters as previously found in C.finmarchicus during its diapause,and most of the storage lipids were used during over-summering.Wax ester polyunsaturated fatty acids(PUFAs) showed the most variation of the fatty acids(FAs),while the percentage composition of FAs in polar lipids was relatively stable.Selective use of wax ester PUFAs has already been shown to play important roles in the winter diapause of Calanus species in other regions,and our FA results show that this is the case for the Yellow Sea Cold Bottom Water(YSCBW) population that diapauses in summer.

Factors aff ecting lipid and fatty acid composition of Calanus sinicus in the Yellow Sea and the East China Sea in spring

The factors aff ecting lipid and fatty acid composition of copepod Calanus sinicus in the Yellow Sea(YS)and the East China Sea(ECS)were examined in this study.In spring,there were signifi cant diff erences between these two regions for both environmental conditions and food availability.Such regional diff erence signifi cantly infl uenced the lipid and fatty acid profi les of C.sinicus.Our results show that C.sinicus has a higher lipids content in ECS,especially for wax ester and triglyceride lipids,indicating a more active and effi cient predation.According to BIO-ENV analysis,the variation of lipids profi les may be infl uenced majorly by water temperature.Moreover,the fatty acids(FAs)profi les of C.sinicus were also diff erent between YS and ECS,especially in the four major contributors,C22꞉1ω11,eicosapentaenoic acid(EPA),docosahexenoic acid(DHA),and C20꞉1ω9.The considerable amounts of self-biosynthesized FAs of herbivorous copepod(C22꞉1ω11 and C20꞉1ω9)and low DHA/EPA ratio may indicate that C.sinicus in ECS feed mainly on phytoplankton comparing to those in YS.The fatty acid profi les of C.sinicus were aff ected by the diff erences in food availability.

Free fatty acids, glucose, and insulin in type 2 diabetes mellitus

Xu et al used the HOMA2 model to estimate theβ-cell function and insulin resistance levels in an individual from simultaneously measured fasting plasma glucose and fasting plasma insulin levels.This method is based on the assumption that the glucose-insulin axis is central for the metabolic activities,which led to type 2 diabetes.However,significant downregulation of both the NKX2-1 gene and the TPD52L3 gene force an increase in the release of free fatty acids(FFAs)into the blood circulation,which leads to a marked reduction in membrane flexibility.These data favor a FFA-glucose-insulin axis.The authors are invited to extend their study with the introduction of the saturation index(number of carbon-carbon double bonds per 100 fatty-acyl chains),as observed in erythrocytes.

Effects of vitamin C deficiency or excess on growth performance, anti-oxidative response and fatty acid composition of juvenile abalone Haliotis discus hannai Ino

A 240-day feeding trial was conducted to investigate the effects of dietary vitamin C on growth performance,anti-oxidative response,and fatty acid composition of juvenile abalone Haliotis discus hannai Ino(initial body weight:0.93±0.00 g).Three semi-purified experimental diets were formulated containing 0.00,94.52,and 9649.58 mg/kg of vitamin C supplied as L-ascorbyl-2-monophosphate.The results show that there was no significant difference in weight gain ratio,daily increment in shell length,and survival rate among the three treatments.Adding dietary vitamin C(9649.58 mg/kg)significantly increased the activities of superoxide dismutase(SOD),glutathine peroxidase(GPX),glitathione-S-transferase(GST),glutathione reductase(GR),alkaline phosphatase(AKP),and lysozyme in viscera(P<0.05).In muscle,activities of phenoloxidase,catalase,SOD,GST,GR,and AKP were increased in the treatment with 9649.58 mg/kg of dietary vitamin C(P<0.05).The highest concentrations of ascorbic acid in viscera and muscle were found in the group with 9649.58 mg/kg of dietary vitamin C(P<0.05).The contents of crude protein and crude lipid in the soft body were significantly increased in the 9649.58 mg/kg group(P<0.05).Dietary vitamin C supplementation significantly decreased the contents of saturated fatty acids(14:0,16:0,and 18:0),and increased the composition of 18:2n-6 and 22:6n-3 in the soft body of abalone(P<0.05).Therefore,although there were no significant effects on the growth performance,dietary vitamin C supplementation improved the anti-oxidation and immune responses,increased specific unsaturated fatty acid(i.e.,16:1,18:1n-7,18:1n-9,18:2n-6 and 22:6n-3),and decreased specific saturated fatty acid(i.e.,14:0,16:0 and 18:0)contents in the soft body of abalone.