HPLC/QTOF-MS metabolomics analysis applied to identify skin biomarkers of UVC-induced skin injury in mice and preventive effects of abietic acid

Abietic acid (AA) is a main constituent from pine resin, which has definite therapeutical effects for treating skin ulcers and tumor. Here, we explored the metabolome changes in skin tissues of mice with UVC-induced skin injury treated with AA by a HPLC-QTOF-MS/MS method. Model mice were induced with UVC irradiation. Skin histopathological changes were examined by routine HE staining. Metabolomic analysis technology and pattern recognition statistical method were applied to analyze the metabolites in the skin tissues of mice to study the therapeutic effect of AA on UVC-induced skin injury in mice. Ceramides, sphingosines, glycyl-L-glutamine, dihydroorotic acid, adenosine, dCMP and lyso-phosphatidylcholines can be used as biomarkers of UVC-induced skin injury. AA can improve the pathological tissue from the pathway of skin lipid and purine pyrimidine metabolism to achieve the therapeutic effect. AA can effectively treat UVC-induced skin injury in mice

Ceramidas Plasmáticas na Estratificação de Risco das Doenças Cardiovasculares / Plasma Ceramides in Cardiovascular Disease Risk Stratification

Resumo A produção de ceramida ocorre em todo o corpo e desempenha um papel importante na manutenção da fisiologia normal. No entanto, os níveis de ceramidas são alterados em estados de doença, principalmente durante o desenvolvimento de diabetes e dislipidemia. A produção de ceramidas também está associada à instabilidade das placas ateroscleróticas. Estudos recentes revelam que pacientes com doença arterial coronariana instável apresentam níveis plasmáticos aumentados de ceramidas (principalmente C16, C18 e C24:1). Atualmente, são consideradas biomarcadores emergentes nas doenças cardiovasculares, sendo utilizadas na predição de instabilidade da placa aterosclerótica e eventos cardiovasculares adversos de forma independente aos fatores de risco tradicionais. Com o objetivo de descrever e discutir o papel das ceramidas na estratificação das doenças cardiovasculares, o desenvolvimento desta revisão narrativa contextualiza a importância desse biomarcador no cenário atual da cardiologia.

Abstract Ceramide production takes place throughout the body and plays a key role in the maintenance of normal physiology. However, ceramide levels are altered during disease states, particularly considering the development of diabetes and dyslipidemia. Ceramide production is also associated with atherosclerotic plaque instability. Recent studies revealed that patients with unstable coronary artery disease (CAD) presented increased plasma ceramide levels (especially C16, C18, and C24:1). These molecules are currently considered emerging biomarkers of cardiovascular diseases (CVD), being used for predicting atherosclerotic plaque instability and adverse cardiovascular events independently from traditional risk factors. With the aim of describing and discussing the role of ceramides in the stratification of cardiovascular diseases, this narrative review contextualizes the importance of this biomarker in the present cardiology scenario.

Research progress of ceramide in cardiovascular diseases / 中国医师杂志

Ceramide is a kind of sphingolipid with complex function, which is involved in cell growth, differentiation, stress response, inflammatory signal transduction and apoptosis as a second messenger. In recent years, many studies have shown that it is closely related to the occurrence and risk prediction of cardiovascular diseases such as coronary heart disease, heart failure, atrial fibrillation, hypertension and so on. In this paper, the related research progress is summarized.

Role of sphingomyelinases in hepatocellular carcinoma / 临床肝胆病杂志

Sphingomyelinases (SMase) are the main enzymes that regulate the signaling pathway of sphingomyelin and the metabolism of related products, and they are involved in the key steps of the complex metabolic process of sphingomyelin. In recent years, many studies have shown that SMase is involved in the biological processes such as cell cycle arrest, cell migration, and inflammation and promotes the development and progression of hepatocellular carcinoma by regulating the apoptosis and proliferation of tumor stem cells. SMase has an important potential biological value in the development, progression, diagnosis, and treatment of hepatocellular carcinoma. This article summarizes the exact role of SMase in the development and progression of hepatocellular carcinoma, in order to provide new ideas and strategies for the clinical treatment of hepatocellular carcinoma and the development of new drugs.

Effect of amitriptyline on lipid deposition and biochemical metabolism in a cell model of nonalcoholic fatty liver disease / 临床肝胆病杂志

ObjectiveTo investigate the effect of amitriptyline on lipid deposition and biochemical metabolism in a cell model of nonalcoholic fatty liver disease (NAFLD) by regulating the acid sphingomyelinase (ASM)/ceramide (CE) pathway. MethodsHepG2 and L02 cells were cultured in vitro to establish a cell model of NAFLD. MTT colorimetry was used to measure cell proliferation rate, and oil red O staining was used to observe the change of lipid droplets in cells. In the experiment, the cells were divided into normal control group, model group, Ami group, TNFα group, and Ami+TNFα group. An automatic biochemical analyzer was used to measure the levels of triglyceride (TG) and total cholesterol (TC) in cells and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in supernatant; ELISA was used to measure the levels of CE and ASM in cells; Western blot was used to measure the protein expression ASM in cells, and RT-PCR was used to measure the mRNA expression of ASM in cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Turkey test was used for further comparison between two groups. ResultsCompared with the normal control group, the NAFLD model group had significant increases in the protein and mRNA expression of ASM and the levels of CE, TG, TC, ALT, and AST (all P＜0.05). Compared with the model group, the Ami group had significant reductions in the protein and mRNA expression of ASM and the levels of CE, TG, TC, ALT, and AST (all P＜0.05), and the TNFα group had significant increases in the protein and mRNA expression of ASM and the levels of CE, TG, ALT, and AST (all P＜0.05). Compared with the TNFα group, the Ami+TNFα group had significant reductions in the protein and mRNA expression of ASM and the levels of CE, TG, TC, ALT, and AST (all P＜0.05). ConclusionThe ASM/CE pathway promotes lipid accumulation and may lead to hepatocyte steatosis, and amitriptyline can alleviate lipid deposition in NAFLD hepatocytes by inhibiting the ASM/CE pathway.

Sphingolipid Mediators of Myocardial Pathology

Cardiomyopathy is the leading cause of mortality worldwide. While the causes of cardiomyopathy continue to be elucidated, current evidence suggests that aberrant bioactive lipid signaling plays a crucial role as a component of cardiac pathophysiology. Sphingolipids have been implicated in the pathophysiology of cardiovascular disease, as they regulate numerous cellular processes that occur in primary and secondary cardiomyopathies. Experimental evidence gathered over the last few decades from both in vitro and in vivo model systems indicates that inhibitors of sphingolipid synthesis attenuate a variety of cardiomyopathic symptoms. In this review, we focus on various cardiomyopathies in which sphingolipids have been implicated and the potential therapeutic benefits that could be gained by targeting sphingolipid metabolism.

Ceramides: Nutrient Signals that Drive Hepatosteatosis

Ceramides are minor components of the hepatic lipidome that have major effects on liver function. These products of lipid and protein metabolism accumulate when the energy needs of the hepatocyte have been met and its storage capacity is full, such that free fatty acids start to couple to the sphingoid backbone rather than the glycerol moiety that is the scaffold for glycerolipids (e.g., triglycerides) or the carnitine moiety that shunts them into mitochondria. As ceramides accrue, they initiate actions that protect cells from acute increases in detergent-like fatty acids; for example, they alter cellular substrate preference from glucose to lipids and they enhance triglyceride storage. When prolonged, these ceramide actions cause insulin resistance and hepatic steatosis, 2 of the underlying drivers of cardiometabolic diseases. Herein the author discusses the mechanisms linking ceramides to the development of insulin resistance, hepatosteatosis and resultant cardiometabolic disorders.

Role of ceramide in development and progression of nonalcoholic fatty liver disease / 临床肝胆病杂志

Nonalcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome caused by liver damage factors except alcohol and has the major feature of diffuse macrovesicular hepatocyte steatosis.Thetwo-hit hypothesis can partly explain the pathogenesis of NAFLD.Recent studies have found that ceramide is a key molecular messenger involved in the development and progression of NAFLD,and as a sphingolipid,it is closely associated with the two-hit hypothesis.This article reviews the role of ceramide in NAFLD.

Role of sphingolipids signaling in pathogenesis of pulmonary fibrosis / 中国药理学与毒理学杂志

Pulmonary fibrosis is a group of chronic lung diseases caused by various factors and characterized by chronic inflammations,lung tissue structure damage,increase of pulmonary interstitial collagen and massive deposition of extracellular matrix (ECM). Because of its complicated etiology, there is no effective treatment currently. Recent studies showed that the activation of sphingolipids signaling and pulmonary fibrosis were closely related. This paper describes the composition and function of sphingolipids signaling pathway and its effect on fibrosis in order to provide new ideas about further study of the pathogenesis of pulmonary fibrosis and methods of prevention.

Induction of a Hardening Phenomenon and Quantitative Changes of Ceramides in Stratum Corneum

BACKGROUND: Hardening phenomenon of human skin after repeated exposure to the irritants is well-known, but the precise mechanism remains elusive. OBJECTIVE: To modify the previous experimental model of hardening phenomenon by repeated applications of two different concentrations of sodium lauryl sulfate (SLS) solutions to Korean healthy volunteers and to investigate the quantitative changes of ceramides in stratum corneum before and after chronic repeated irritation. METHODS: Eight hundred microliters of distilled water containing 0.1% and 2% SLS was applied for 10 minutes on the forearm of 41 healthy volunteers for 3 weeks. After an intervening 3-week rest, 24-hour patch tests with 1% SLS were conducted on previously irritated sites. Transepidermal water loss (TEWL), erythema index and quantity of ceramide were measured in the stratum corneum before and after irritation. RESULTS: TEWL values on the sites preirritated with 2% SLS were lower than those with 0.1% SLS. Hardening phenomenon occurred in 24 volunteers at day 44. The changes in ceramide levels were not significantly higher in the hardened skin than in the non-hardened skin. CONCLUSION: Repetitive stimulation with a higher concentration of SLS can more easily trigger skin hardening.

LASS family and tumor / 国际肿瘤学杂志

LASS gene family is a group of highly conserved and longevous gene LASS protein has a ceramide synthase activity,which can synthesize different types of ceramides.Ceramide is a key intermediate in tumor cells growth,signal transduction and other aspects.Thus it is inferred that the expression of LASS gene can suppress the occurrence and development of tumor within a certain range.

Ceramides and terpenoids from Russula austrodelica Singer / Ceramidas y terpenoides desde Russula austrodelica Singer

A mixture of ceramides and known terpenes, was obtained from the fruiting bodies of Russula austrodelica. The structures were determined from chemical and spectroscopic evidence. R. austrodelica is a mycorrhizal fungus that grow in the Nothophagus forests of southern Chile. This is the first report of the isolation of ceramides in Chilean mushrooms.

Una mezcla de ceramidas y de terpenos conocidos, se obtuvo de los cuerpos fructíferos de Russula austrodelica. Las estructuras fueron determinadas a partir de evidencias químicas y espectroscópicas. R. austrodelica es un hongo micorrícico que crecen en los bosques de Nothophagus del sur de Chile. Este es el primer informe del aislamiento de ceramidas en hongos chilenos.

Effects of ceramide-induced apoptosis on oncotherapy / 国际肿瘤学杂志

Ceramide (Cer),as the center molecular of sphingolipids metabolism,functions as a bioactive mediator of different cellular processes,such as cell growth,differentiation,senescence and apoptosis,besides being structural component of cellular membrane.It can inhibit cell proliferation and promote cell apoptosis via multiple signal pathawys.Chemicals anti-cancer properties based on Cer metabolism that upregulate its level are confirmed in plenty of experimental researches,and parts of research productions are applied in anticancer clinic experiments.

Effect of ceramide on peritoneal mesothelial cells apoptosis induced by peritoneal dialysis solution / 中华肾脏病杂志

ObjectiveToexploretheeffectof ceremideonprocess of peritoneal mesothelial cells(PMCs) apoptosis induced by peritoneal dialysis solution(PDS).Methods PMCs were cultured with normal DMEM,1.5％ PDS and 4.25％ PDS.4.25％ mannitol was used as high osmotic pressure control.Ceremide were detected by LC-MS-MS.Flow cytometry was used in apoptosis analysis.Bax,p53 and bcl-2 protein expressions were detected by Western blotting.Results (1) PDS caused the increase of intracellular ceremide in PMCs,and normal and high osmotic pressure controls had no such effect.As the acidic sphigomyelinase inhibitor,desipramine significantly inhibited the production of ceramide induced by 4.25％ PDS [(56.08±12.24) μg/L vs (91.25:t:15.89) μg/L,P＜0.01]. (2) Compared with 1.5％ PDS,4.25％ PDS stimulated PMCs apoptosis (26.65％±6.21％ vs 4.04％±1.86％,P＜0.01),up-regulated bax and p53 proteins expression (P＜0.01),and down-regulated bcl-2 protein exprssion(P＜0.05).Desipramine obviously inhibited the apoptosis induced by 4.25％ PDS,decreased bax and p53 proteins expression,increased bcl-2 protein expression(P＜0.05).Exogenous C2-ceremide reversed the effect of desipramine(P＜0.05).Conclusion The increase of intracellular ceremide may play an important role in the PMCs apoptosis induced by high glucose PDS.

Effects of Prinsepia utilis Royle oil on the synthesis of ceramide and expression of ceramidase / 中华皮肤科杂志

Objective To evaluate the effects of Prinsepia utilis Royle oil (PURO) on the synthesis of ceramide and expression of acid ceramidase N-acylsphingosine amidohydrolase 1 (ASH1),and to explore the mechanisms underlying its moisturizing and skin barrier-repairing effects.Methods Keratinocytes from human foreskin tissue were classified into 2 groups to be cultured in keratinocyte-serum free medium (K-SFM) with or without the presence of PURO.Enzyme linked immunosorbent assay (ELISA) was performed to measure the level of ceramide in the culture supernatant of keratinocytes at 0,3,8,24 and 48 hours.The back of nude mice was divided into 4 areas,i.e.,test area,matrix area,blank control area and negative control area.Acetone and ether were used to destroy the epidermal barrier in the test,matrix,and blank control areas,then,the former 2 areas were topically treated with emulsions containing 1％ PURO and matrix,respectively,and the blank control area remained untreated.The epidermal barrier remained intact and untreated in the negative control area.Noninvasive methods were used to determine transepidermal water loss (TEWL),epidermal moisture content and skin lipid content in these areas on day 0,1,3,and 7.Skin tissue was obtained from these areas on day 0 and 7 followed by an immunohistochemical study for the quantification of ASH1 expression.Results The level of supernatant ceramide increased with time in the PURO-treated keratinocytes,which was significantly higher at 24 hours and 48 hours than at 0 hour (1.3817 ± 0.100 and 1.3737 ± 0.047 vs.0.7630 ± 0.143,both P ＜ 0.05).The supernatant ceramide was also elevated in the PURO-treated keratinocytes compared with untreated keratinocytes at 24 and 48 hours (both P ＜ 0.05).Noninvasive skin tests showed a gradual decrease in the TEWL,but an increase in the epidermal moisture content and skin lipid content with time in the 3 epidermal barrier-destroyed areas.As far as the test area was concerned,TEWL value was significantly lower on day 3 and 7 than on day 0 (10.85 ± 0.64 and 8.01 ± 0.58 vs.12.65 ± 0.71,both P ＜ 0.05),while a significant increment was observed in the skin lipid content on day 3 and 7 compared with day 0 (29.14 ± 0.40 and 31.30 ± 0.88 vs.27.02 ± 0.65,both P ＜ 0.05),as well as in the epidermal moisture content on day 1,3 and 7 compared with day 0 (13.98 ± 0.28,15.00 ± 0.38 and 15.86 ± 0.18 vs.11.74 ± 0.62,all P＜ 0.05).On day 7,there was a statistical decline in TEWL value,but an elevation in epidermal moisture content,skin lipid content and ASH1 expression in the test area compared with the matrix area and blank control area (all P ＜ 0.05).Also,the expression of ASH1 was upregulated on day 7 compared with day 0 in the 3 barrier-destroyed areas (all P ＜ 0.05).Conclusion PURO may exert skin-moisturizing and barrier-repairing effects by enhancing the synthesis of ceramide and expression of acid ceramidase ASH1.

Studies on chemical constituents of Anthogorgia sp / 第二军医大学学报

Objective: To study the chemical constituents of Anthogorgia sp. collected from Beihai area of Guangxi Zhuang Autonomous Region, China. Methods The Et 2O extract of Anthogorgia sp. was subjected to repeated silica gel (eluted with Pe: Ac=99:1, 49:1, 39:1, 34:1, 29:1, 24:1, 19:1, 14:1, 11:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 100%Ac; CHCl3:MeOH=24:1, 19:1, 14:1, 9:1, 7:1, 5:1, 3:1, 1:1, 1:4, 1:9, 1:39, 100% MeOH) and Sephadex LH-20 column chromatography (n-hexane:CHCl3:MeOH=2:1:1) for isolation and purification. The structures of the resultant compounds were elucidated using spectroscopic analysis (1 H NMR, 13C NMR and MS). Results: Six steroids and one ceramide were isolated and identified as: Ergost-5, 24(28)-dien-3β-ol(1),(22E,24R)-ergost-7,22-dien-3β,5α, 6β-triol(2),(22E,24S)-ergost-5,22-dien-3β-ol(3), cholesta-5-en- 3β,7β,19α-triol(4),(22E)-cholesta-5,22-dien-3β-ol(5), cholesterol(6), and N-palmitoyloctadecasphinga-4(E)-ene(7). Conclusion: All the six compounds are isolated from Anthogorgia sp. for the first time.

Effects of Endurance Exercise and High-Fat Diet on Insulin Resistance and Ceramide Contents of Skeletal Muscle in Sprague-Dawley Rats / 당뇨병

BACKGROUND: We evaluated the effects of endurance exercise and a high-fat diet on insulin resistance and ceramide contents of skeletal muscle in Sprague-Dawley rats. METHODS: We randomly divided 32 rats into four groups: control (CON, n = 8), high fat diet (HF, n = 8), exercise (Ex, 24 m/min for 2 hours, 5 days/wk, n = 8), HF/Ex (n = 8). After 4-week treatments, plasma lipid profiles, glucose and insulin concentrations were measured. The triglycerides (TG), ceramide, and glucose transporter 4 (GLUT-4) contents were measured in the skeletal muscle. The rate of glucose transport was determined under submaximal insulin concentration during the muscle incubation. RESULTS: Free fatty acid levels were significantly higher in CON and HF than Ex (P = 0.032). Plasma glucose levels in HF were significantly higher than the two Ex groups (P = 0.002), and insulin levels were significantly higher in HF than in other three groups (P = 0.021). Muscular TG concentrations were significantly higher in HF than CON and Ex and also in HF/Ex than Ex, respectively (P = 0.005). Hepatic TG concentrations were significantly higher in HF than other three groups but Ex was significantly lower than HF/Ex (P = 0.000). Muscular ceramide content in HF was significantly greater than that in either Ex or HF/Ex (P = 0.031). GLUT-4 levels in CON and HF were significantly lower than those in Ex and HF/Ex (P = 0.009, P = 0.003). The glucose transport rate in submaximal insulin concentration was lower in CON than in either Ex or HF/Ex (P = 0.043), but not different from HF. CONCLUSION: This study suggests that high fat diet for 4 weeks selectively impairs insulin resistance, but not glucose transport rate, GLUT-4 and ceramide content in skeletal muscle per se. However, endurance exercise markedly affects the content of ceramide and insulin resistance in muscle.

Changes of Antimicrobial Peptides and Transepidermal Water Loss After Topical Application of Tacrolimus and Ceramide-dominant Emollient in Patients with Atopic Dermatitis

Increased transepidermal water loss (TEWL) and downregulated antimicrobial peptides (AMPs) are observed in patients with atopic dermatitis (AD). Tacrolimus and ceramide-dominant emollients are effective in the treatment of AD by preventing the production of inflammatory cytokines and by correcting skin barrier dysfunctions, respectively. Present study was designed to investigate the relationship between antimicrobial and barrier factors by measuring the changes of AMPs and TEWL after topical application of tacrolimus and ceramide-dominant emollient in the patients with AD. A total of three patients with AD were treated with tacrolimus in one lesion and ceramide-dominant emollient in another lesion for 4 weeks. RT-PCR and western blotting revealed that the mRNA and protein expression levels of hBD-2 and LL-37 were increased on the both study sites. Immunohistochemical analysis showed significant increase of AMPs and IL-1alpha, while, IL-4 was decreased on the both study sites. The mean changes of TEWL and AMPs showed no statistical difference between both sites. Tacrolimus and ceramide-dominant emollient influence on both TEWL and AMPs expression in patients with AD, namely they have similar effects on both of the two. This study shows that restoration of permeability barrier function is accompanied by the concomitant improvement of antimicrobial defense in patients with AD.

Metabolites of sphingolipids as targets for cancer therapy / 军事医学科学院院刊

Sphingolipids are structural components of cellular membranes.Their metabolites,such as ceramide,sphingosine,and sphingosine 1-phosphate are important bioactive molecules,which act as the first or second messengers regulating various cellular activities,including proliferation,survival,migration and neovascularization in vitro and in vivo tumor models.Sphingosine kinases 1(SPK1) is a critical regulator of the balance between ceramide and S1P.Recent studies show that ceramide,SPK1 and S1P can regulate many of the hallmarks of cancer.These studies suggest that these metabolites can serve as novel targets for cancer therapy.

Relationship between ceramide, gastrins and colorectal neoplasms / 国际外科学杂志

It is confirmed that expression of gastrins in some colorectal neoplasms cells and their tissue is abnormal. The abnormal expression of gastrins have close relationship with the occurrence and the develop-ment of colorectal neoplasms. Gastrins binding with receptor can regulate ceramides content and promote cells proliferation by series of signal transduction pathways. Blocking these signal transduction pathways may provide a novel therapeutic approach for colorectal neoplasms prevention and treatment.