AIMTo study the chemical constituents of Hemistepta lyrata.

METHODSTo separate compounds with various chromatography technology and to elucidate their structures by chemical and spectral analysis.

RESULTSTwo compounds were isolated from Hemistepta lyrata and their structures were determined as hemiceramide (I), hemisterpene ether (II).

CONCLUSIONCompounds I and II are new compounds.

Asteraceae ; chemistry ; Ceramides ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo establish a high-performance liquid chromatography-tandem mass spectrometry (HPLC-ESI-MS) method for simultaneous quantitative analysis of different ceramide species in cells.

METHODSThe analysis was performed on an Agilent 1290 HPLC system with a ZORBAX Eclipse XDB-C8 (150.0 mm × 2.1 mm, 3.5 μL) column and a temperature of 35 ℃. Methanol with 1 mmol/L ammonium formate and 0.2% formic acid was used as mobile phase A and 100% methanol was used as mobile phase B. And the ceramides were separated by gradient elution at a flow rate of 0.3 mL/min. Electrospray ionization (ESI) with multiple reaction monitoring (MRM) was used in the analysis.

RESULTSFour ceramide species all had a good linear response in the determination ranges (R² ≥ 0.9987). The average recoveries (n = 9) were 99.1%,99.9%,100.5% and 98.2% with RSDs of 5.6%, 5.1%, 4.7% and 5.5%, respectively. In addition, the levels of ceramides in FL cells were relatively stable, while the C24-ceramide had the highest level.

CONCLUSIONThe HPLC-ESI-MS method for simultaneous analysis of ceramides has high accuracy, reproducibility and linearity, which may be used for quantification of ceramide species in cells.

Ceramides ; chemistry ; Chromatography, High Pressure Liquid ; Reproducibility of Results ; Tandem Mass Spectrometry

AIMTo study the bioactive constituents from Anthopleura pacifica.

METHODSCompounds were separated by Pyricularia oryzae bioassay-guided fractionation method with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and chemical evidence.

RESULTSA portion showing activity against P. oryzae was obtained and from the portion four compounds were identified as N-hydroxyethyl-N-tetradecanoyl-(2S,3R)-octadecasphinga-4(E), 8(E)-dienine (a), N-hydroxyethyl-N-(9Z-hexadecenoyl)-(2S,3R)-octadecasphinga-4 (E), 8 (E)-dienine (b), N-hydroxyethyl-N-hexadecanoyl-(2S,3R)-octadecasphinga-4(E), 8 (E)-dienine (c) and N-hydroxyethyl-N-(13Z-docosenoyl-(2S,3R)-octadecasphinga-4(E), 8(E)-dienine(d).

CONCLUSIONAll the four compounds are new ceramides.

Animals ; Biological Assay ; Ceramides ; chemistry ; isolation & purification ; Cnidarian Venoms ; chemistry ; Mitosporic Fungi ; drug effects ; Molecular Conformation ; Molecular Structure ; Sea Anemones ; chemistry

OBJECTIVETo observe the effects of antioxidation and ceramide content of improved prescription of Didang-tang (IPDT) on exprimental atherosclerosis(AS) rabbits.

METHODPlasm Superoxide Dismutase(SOD) activity was detected with micro-content fast detecting method, Plasm Malondialdehyde(MDA) content with improved BaMuGuoFu method, and Aortic Ceramide (CER) content with thinlayer scanning.

RESULTIPDT could effectivly improve plasma SOD activity and decrease plasma MDA content and decrease aortic CER content.

CONCLUSIONIPDT on exprimental AS is related to the improvement of antioxidation and decrease of CER content.

Animals ; Antioxidants ; pharmacology ; Arteriosclerosis ; metabolism ; Ceramides ; metabolism ; Cinnamomum ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Leeches ; chemistry ; Male ; Materia Medica ; pharmacology ; Plants, Medicinal ; chemistry ; Rabbits ; Rheum ; chemistry

OBJECTIVETo search for new antitumor active lead compounds from marine microorganism.

METHODA marine strain, Aspergillus terreus, was cultured and up-scaled in artificial seawater media, from which the metabolites were isolated and elucidated by using modern spectroscopy techniques.

RESULTTwelve compounds were isolated from mycelia and fermentation broth of A. terreus.

CONCLUSIONCompounds 1-4 were steroids, compounds 5-8 were organic acids and esters, compound 9 was an alkaloid, compound 10 was an isocoumarin, compound 11 was ceramide, compound 12 was propenyl cyclic pentanediol.

Alkaloids ; chemistry ; isolation & purification ; metabolism ; Aspergillus ; chemistry ; metabolism ; Ceramides ; chemistry ; isolation & purification ; metabolism ; Culture Media ; chemistry ; metabolism ; Esters ; chemistry ; isolation & purification ; metabolism ; Isocoumarins ; chemistry ; isolation & purification ; metabolism ; Mycelium ; chemistry ; metabolism ; Propylene Glycols ; chemistry ; isolation & purification ; metabolism ; Rhizophoraceae ; microbiology ; Steroids ; chemistry ; isolation & purification ; metabolism

Ceramides are the main lipid component maintaining the lamellae structure of stratum corneum, as well as lipid second messengers for the regulation of cellular proliferation and/or apoptosis. In our previous study, psoriatic skin lesions showed marked decreased levels of ceramides and signaling molecules, specially protein kinase C-alpha (PKC-alpha) and c-jun N-terminal kinase (JNK) in proportion to the psoriasis area and severity index (PASI) scores, which suggested that the depletion of ceramide is responsible for epidermal hyperproliferation of psoriasis via downregulation of proapoptotic signal cascade such as PKC-alpha and JNK. In this study, we investigated the protein expression of serine palmitoyltransferase (SPT) and ceramidase, two major ceramide metabolizing enzymes, in both psoriatic epidermis and non-lesional epidermis. The expression of SPT, the ceramide generating enzyme in the de novo synthesis in psoriatic epidermis, was significantly less than that of the non-lesional epidermis, which was inversely correlated with PASI score. However, the expression of ceramidase, the degradative enzyme of ceramides, showed no significant difference between the lesional epidermis and the non-lesional epidermis of psoriatic patients. This might suggest that decreased expression of SPT protein is one of the important causative factors for decreased ceramide levels in psoriasis.
Adolescent ; Adult ; Amidohydrolases/*biosynthesis/metabolism ; Apoptosis ; Cell Proliferation ; Ceramidases ; Ceramides/chemistry ; Child ; Epidermis/metabolism ; Female ; Humans ; JNK Mitogen-Activated Protein Kinases/metabolism ; Male ; Models, Biological ; Protein Kinase C-alpha/metabolism ; Psoriasis/*blood/diagnosis ; Serine C-Palmitoyltransferase/*biosynthesis

In the present study, a new ceramide, namely 2S, 3R-4E, 8E-2-(heptadecanoylamino)-heptadeca-4, 8-diene-1, 3-diol (1), along with four known steroids, including 24-methylcholesta-5, 24(28)-diene-3β-ol (2), 24-methylcholesta-5, 24(28)-diene-3β-acetate (3), 4-methyl-24-methylcholesta-22-ene-3-ol (4), and cholesterol, was isolated and characterized from CHCl/MeOH extract of Cespitularia stolonifera. A new acetate derivative of compound 1, termed 2S, 3R-4E, 8E-2-(heptadecanoylamino)-heptadeca-4, 8-diene-1, 3-diacetate (1a), was also prepared in the present study. All the structures were established on the basis of modern spectroscopic techniques, including FT-IR, 1D, 2D-NMR, HRESI-MS, and GC-MS, in addition of chemical methods. (-)-Alloaromadendren, ledane, (1)-alloaromadendren oxide, isoaromadendrene epoxide and (-)-caryophellen oxide were identified from the n-hexane fraction using GC-MS. The extract and the two ceramides (1) and (1a) exhibited significant cytotoxic activity against lung cancer A549 cells, while the extract and the two steroids (2) and (3) exhibited significant cytotoxic activity against breast cancer MCF-7 cells. The CHCl/MeOH extract exhibited significant antiulcer activity in both ethanol and acetic acid induced ulcer models in rats, as evidenced by histopathological, histochemical, and biochemical examinations.
A549 Cells ; Acetic Acid ; Animals ; Anthozoa ; chemistry ; Anti-Ulcer Agents ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Biological Products ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Breast Neoplasms ; drug therapy ; Ceramides ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Disease Models, Animal ; Ethanol ; Female ; Humans ; Lung Neoplasms ; drug therapy ; MCF-7 Cells ; Magnetic Resonance Spectroscopy ; methods ; Rats ; Spectroscopy, Fourier Transform Infrared ; methods ; Steroids ; chemistry ; isolation & purification ; pharmacology ; therapeutic use ; Ulcer ; chemically induced ; drug therapy

In humans, skin barrier dysfunction is thought to be responsible for enhanced penetration of allergens. Similar to conditions seen in humans, canine atopic dermatitis (CAD) is characterized by derangement of corneocytes and disorganization of intercellular lipids in the stratum corenum (SC) with decreased ceramide levels. This study was designed to evaluate the effects of a moisturizer containing ceramide on dogs with CAD. Dogs (n = 20, 3~8 years old) with mild to moderate clinical signs were recruited and applied a moisturizer containing ceramide for 4 weeks. Transepidermal water loss (TEWL), skin hydration, pruritus index for canine atopic dermatitis (PICAD) scores, and canine atopic dermatitis extent and severity index (CADESI) scores of all dogs were evaluated. Skin samples from five dogs were also examined with transmission electron microscopy (TEM) using ruthenium tetroxide. TEWL, PICAD, and CADESI values decreased (p < 0.05) and skin hydration increased dramatically over time (p < 0.05). Electron micrographs showed that the skin barrier of all five dogs was partially restored (p < 0.05). In conclusion, these results demonstrated that moisturizer containing ceramide was effective for treating skin barrier dysfunction and CAD symptoms.
Animals ; Ceramides/*therapeutic use ; Cholesterol/*therapeutic use ; Dermatitis, Atopic/complications/drug therapy/physiopathology/*veterinary ; Dog Diseases/*drug therapy/etiology/physiopathology ; Dogs ; Emollients/*therapeutic use ; Epidermis/drug effects/physiopathology/ultrastructure ; Fatty Acids, Nonesterified/*therapeutic use ; Female ; Male ; Microscopy, Electron, Transmission/veterinary ; Pruritus/drug therapy/etiology/physiopathology/veterinary ; Republic of Korea ; Ruthenium Compounds/chemistry ; Water Loss, Insensible/drug effects

Abstract Phospholipase D (PLD) plays an important role as an effector in a variety of physiological processes that reveal it to be a member of the signal transducing phospholipases. Recently, PLD2 was reported as a necessary intermediate in preventing apoptosis induced by hydrogen peroxide or hypoxia in rat pheochromocytoma (PC12) cells. The data presented here show that both PLD isozymes, PLD1 and PLD2 are also required in attenuating glutamate-induced cell death in PC12 cells. Treatment of PC12 cells with glutamate resulted in induction of apoptosis in these cells, which is accompanied by decreased PLD activity and increased ceramide concentration. Incubation of PC12 cells with exogenous C6-ceramide showed a time-dependent decrease of PLD activity. When cDNAs of PLD1 and PLD2 were transfected into PC12 cells respectively, overexpression of PLD1 or PLD2 resulted in inhibition of glutamate-induced apoptotic cell death. These data indicate that both PLD1 and PLD2 play a protective role against glutamate-induced cell death in PC12 cells.
Animals ; Apoptosis/drug effects/*physiology ; Cell Survival/drug effects ; Ceramides/pharmacology ; Dose-Response Relationship, Drug ; Enzyme Activation ; Gene Expression Regulation, Enzymologic/drug effects ; Glutamic Acid/*toxicity ; Isoenzymes/drug effects/genetics/*metabolism ; Kinetics ; PC12 Cells ; Phospholipase D/chemistry/drug effects/genetics/*metabolism ; Rats ; Sphingolipids/metabolism