2.Advance in research on regulatory mechanism and functions of neutral sphingomyelinse 2.
Acta Academiae Medicinae Sinicae 2013;35(5):581-585
Neutral sphingomyelinase 2 (nSMase2), which located mainly on the plasma membrane, hydrolyzes sphingomyelin into ceramide and plays an important role in the physiological and pathological regulation of cell apoptosis, cell growth arrest, and inflammation. nSMase2 is also involved in the development of Alzheimer's disease and the bone growth.Under neutral pH and the presence of Ca(2+), Mg(2+), and Mn(+), the activity of nSMase2 is induced by oxidative stress through phosphorylation. Furthermore, the induced interaction of anionic phospholipids and the signaling molecules like receptor for activated C-kinase 1/embryonic ectodermal development with nSMase2 are also crucial mechanisms of protein activation. In the review, recent research advances in the structure and function of nSMase2 and its underlying mechanisms are summarized.
Animals
;
Humans
;
Molecular Structure
;
Signal Transduction
;
Sphingomyelin Phosphodiesterase
;
chemistry
;
physiology
3.Ceramide is Involved in MPP+ -induced Cytotoxicity in Human Neuroblastoma Cells.
Eun Joo NAM ; Hye Sook LEE ; Young Jae LEE ; Wan Seok JOO ; Sungho MAENG ; Hye In IM ; Chan Woong PARK ; Yong Sik KIM
The Korean Journal of Physiology and Pharmacology 2002;6(6):281-286
To understand the cytotoxic mechanism of MPP+, we examined the involvement of ceramide in MPP+ -induced cytotoxicity to human neuroblastoma SH-SY5Y cells. When SH-SY5Y cells were exposed to MPP+, MPP+ induced dose-dependent cytotoxicity accompanied by 2-fold elevation of intracellular ceramide levels in SH-SY5Y cells. Three methods were used to test the hypothesis that the elevated intracellular ceramide is related to MPP+ -induced cytotoxicity: C2-ceramide was directly applied to cells, sphingomyelinase (SMase) was exogenously added, and oleoylethanolamine (OE) was used to inhibit degradation of ceramide. Furthermore, inhibition of ceramide-activated protein phosphatase (CAPP), the effector of ceramide, using okadaic acid (OA) attenuated cell death but treatment of fumonisin B1, the ceramide synthase inhibitor, did not alter the cytotoxic effect of MPP+. Based on these, we suggest that the elevation of intracellular ceramide is one of the important mediators in MPP+ -induced cell death.
Cell Death
;
Humans*
;
Neuroblastoma*
;
Okadaic Acid
;
Sphingomyelin Phosphodiesterase
4.Vitamin C Stimulates Epidermal Ceramide Production by Regulating Its Metabolic Enzymes.
Kun Pyo KIM ; Kyong Oh SHIN ; Kyungho PARK ; Hye Jeong YUN ; Shivtaj MANN ; Yong Moon LEE ; Yunhi CHO
Biomolecules & Therapeutics 2015;23(6):525-530
Ceramide is the most abundant lipid in the epidermis and plays a critical role in maintaining epidermal barrier function. Overall ceramide content in keratinocyte increases in parallel with differentiation, which is initiated by supplementation of calcium and/or vitamin C. However, the role of metabolic enzymes responsible for ceramide generation in response to vitamin C is still unclear. Here, we investigated whether vitamin C alters epidermal ceramide content by regulating the expression and/or activity of its metabolic enzymes. When human keratinocytes were grown in 1.2 mM calcium with vitamin C (50 mug/ml) for 11 days, bulk ceramide content significantly increased in conjunction with terminal differentiation of keratinocytes as compared to vehicle controls (1.2 mM calcium alone). Synthesis of the ceramide fractions was enhanced by increased de novo ceramide synthesis pathway via serine palmitoyltransferase and ceramide synthase activations. Moreover, sphingosine-1-phosphate (S1P) hydrolysis pathway by action of S1P phosphatase was also stimulated by vitamin C supplementation, contributing, in part, to enhanced ceramide production. However, activity of sphingomyelinase, a hydrolase enzyme that converts sphingomyelin to ceramide, remained unaltered. Taken together, we demonstrate that vitamin C stimulates ceramide production in keratinocytes by modulating ceramide metabolic-related enzymes, and as a result, could improve overall epidermal barrier function.
Ascorbic Acid*
;
Calcium
;
Epidermis
;
Humans
;
Hydrolysis
;
Keratinocytes
;
Serine C-Palmitoyltransferase
;
Sphingomyelin Phosphodiesterase
;
Vitamins*
5.Neutral Sphingomyelinase and Breast Cancer Research.
Sun Yong HWANG ; Tae Hee KIM ; Hae Hyeog LEE
Journal of Menopausal Medicine 2015;21(1):24-27
Our understanding of the functions of neutral sphingomyelinase (N-SMase) signaling has advanced over the past decade. In this review, we focus on the roles and regulation of N-SMase 1, N-SMase 2, N-SMase 3, an enzyme that generates the bioactive lipid ceramide through the hydrolysis of the membrane lipid sphingomyelin. A large body of work has now implicated N-SMase 2 in a diverse set of cellular functions, physiological processes, and disease pathologies. We focus on different aspects of this enzyme's regulation from transcriptional, post-translational, and biochemical. Furthermore, we expected N-SMase involvement in cellular processes including inflammatory signaling, cell growth, apoptosis, and tumor necrosis factor which in turn play important roles in pathologies such as cancer metastasis, variable disease, and other organ system disorders. Lastly, we examine avenues where targeted N-SMase inhibition may be clinically beneficial in disease scenarios.
Apoptosis
;
Breast Neoplasms*
;
Hydrolysis
;
Membranes
;
Neoplasm Metastasis
;
Pathology
;
Physiological Processes
;
Sphingomyelin Phosphodiesterase*
;
Tumor Necrosis Factor-alpha
6.A novel SMPD1 mutation in two Chinese sibling patients with type B Niemann-Pick disease.
Rong HUA ; Hui WU ; Zhe CUI ; Jin-xian CHEN ; Zheng WANG
Chinese Medical Journal 2012;125(8):1511-1512
Type B Niemann-Pick disease is an autosomal recessive sphingolipidosis due to mutations in the sphingomyelin phosphodiesterase 1 gene (SMPD1). Here we present molecular findings for two sibling patients. One mutation V36A due to c.107T>C in exon 1 is a single nucleotide polymorphism and the other N522S due to c.1565 A>G in exon 6 is a novel missense mutation. This non-fatal missense mutation leads to –20% residual lysosomal acid sphingomyelinase activity in vitro and only results in hepatosplenomegaly without neurologic involvement.
Female
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Humans
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Middle Aged
;
Mutation, Missense
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Niemann-Pick Disease, Type B
;
genetics
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Polymorphism, Single Nucleotide
;
Siblings
;
Sphingomyelin Phosphodiesterase
;
genetics
7.Expression of ASMase in alcoholic liver fibrosis in rats.
Mi WANG ; Qin-fang CAO ; Ping LIU ; Xiao-dan LU ; Shu-juan ZHANG ; Wang-xian TANG ; Cui-huan WU
Chinese Journal of Hepatology 2013;21(12):920-923
OBJECTIVETo investigate the expression of the lysosomal enzyme acid sphingomyelinase (ASMase) in alcohol-induced hepatic fibrosis using a rat model.
METHODSThe model of liver fibrosis was induced by administration of alcohol and high fat diet using 20 rats. Six rats given no alcohol and normal diet served as the control group. Real-time PCR, western blotting, and immunohistochemistry were used to evaluate fibrosis-related changes in the mRNA and protein expressions of ASMase.
RESULTSThe fibrotic liver tissues of the model rats showed significantly higher expression levels of ASMase than the non-fibrotic liver tissues of the control rats (P less than 0.05).
CONCLUSIONExpression of ASMase is increased in the fibrotic liver tissue of an alcohol-induced hepatic fibrosis rat model, suggesting that this lysosomal enzyme may contribute to development of this disease condition.
Animals ; Liver ; enzymology ; Liver Cirrhosis, Alcoholic ; enzymology ; Liver Cirrhosis, Experimental ; enzymology ; Male ; Rats ; Rats, Sprague-Dawley ; Sphingomyelin Phosphodiesterase ; metabolism
8.Role of sphingomyelin-MAPKs pathway in heat-induced apoptosis.
Hyun Sook CHUNG ; So Ra PARK ; Eun Kyung CHOI ; Heon Jin PARK ; Robert J GRIFFIN ; Chang W SONG ; HeonJoo PARK
Experimental & Molecular Medicine 2003;35(3):181-188
The role of sphingomyelinase (SMase) activation and mitogen activated protein kinases (MAPKs) activation in cellular apoptosis was investigated during the hyperthermic treatment of HL-60 human leukemia cells. Treating the cells for 1 h at 43oC caused more than 50% of cellular apoptosis within several hours. The neutral-SMase activity in the cells treated for 1 h at 42degrees C was slightly increased but decreased in the cells treated at 43degrees C or 44degrees C for the same period whereas the acid SMase activity was slightly increased after heating the cells at 42degrees C and 43degrees C and markedly increased at 44degrees C for 1 h. Treatment of cells with inhibitors of SMase activation and ceramide formation significantly reduced the heat-induced apoptosis. Three major families of mitogen-activated protein kinases (MAPKs) i.e. ERK1/2, p38 and JNK, were activated by the hyperthermic treatment of cells. Inhibition of ERK1/2 with PD98059 exerted little effect on the heat-induced apoptosis and p38 inhibition with SB203580 slightly lessened apoptosis whereas, inhibition of JNK with SP600125 markedly suppressed the heat-induced apoptosis. These results indicate that heat-shock induced the activation of SMase, particularly acid-SMase, thereby causing apoptosis and that JNK played a pivotal role in heat-induced apoptosis in HL-60 leukemia cells.
Apoptosis/*physiology
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Enzyme Activation
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HL-60 Cells
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Heat
;
Human
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Mitogen-Activated Protein Kinase Kinases/*metabolism
;
Sphingomyelin Phosphodiesterase/*metabolism
9.Comparative effect of dietary borage oil and safflower oil on anti-proliferation and ceramide metabolism in the epidermis of essential fatty acid deficient guinea pigs.
Journal of Nutrition and Health 2015;48(4):319-326
PURPOSE: Borage oil (BO) and safflower oil (SO) are efficacious in reversing epidermal hyperproliferation, which is caused by the disruption of epidermal barrier. In this study, we compared the antiproliferative effect of dietary BO and SO. Altered metabolism of ceramide (Cer), the major lipid of epidermal barrier, was further determined by measurement of epidermal levels of individual Cer, glucosylceramide (GlcCer), and sphingomyelin (SM) species, and protein expression of Cer metabolizing enzymes. METHODS: Epidermal hyperproliferation was induced in guinea pigs by a hydrogenated coconut diet (HCO) for 8 weeks. Subsequently, animals were fed diets of either BO (group HCO + BO) or SO (group HCO + SO) for 2 weeks. As controls, animals were fed BO (group BO) or HCO (group HCO) diets for 10 weeks. RESULTS: Epidermal hyperproliferation was reversed in groups HCO + BO (67.6% of group HCO) and HCO + SO (84.5% of group HCO). Epidermal levels of Cer1/2, GlcCer-A/B, and beta-glucocerebrosidase (GCase), an enzyme of GlcCer hydrolysis for Cer generation, were higher in group HCO + BO than in group HCO, and increased to levels similar to those of group BO. In addition, epidermal levels of SM1, serine palmitoyltransferase (SPT), and acidic sphingomyelinase (aSMase), enzymes of de novo Cer synthesis and SM hydrolysis for Cer generation, but not of Cer3-7, were higher in group HCO + BO than in group HCO. Despite an increase of SPT and aSMase in group HCO + SO to levels higher than in group HCO, epidermal levels of Cer1-7, GlcCer-A/B, and GCase were similar in these two groups. Notably, acidic ceramidase, an enzyme of Cer degradation, was highly expressed in group HCO + SO. Epidermal levels of GlcCer-C/D and SM-2/3 did not differ among groups. CONCLUSION: Dietary BO was more prominent for reversing epidermal hyperproliferation by enhancing Cer metabolism with increased levels of Cer1/2, GlcCer-A/B, and SM1 species, and of GCase proteins.
Animals
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Borago*
;
Carthamus tinctorius*
;
Ceramidases
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Cocos
;
Diet
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Epidermis*
;
Glucosylceramidase
;
Guinea Pigs*
;
Guinea*
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Hydrogen
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Hydrolysis
;
Metabolism*
;
Safflower Oil*
;
Serine C-Palmitoyltransferase
;
Sphingomyelin Phosphodiesterase
10.Analysis of Acid Sphingomyelinase Activity in Dried Blood Spots Using Tandem Mass Spectrometry.
Elisa LEGNINI ; Joe J ORSINI ; Adolf MUHL ; Britt JOHNSON ; Angela DAJNOKI ; Olaf A BODAMER
Annals of Laboratory Medicine 2012;32(5):319-323
BACKGROUND: Niemann Pick disease (NP) is a rare, lysosomal storage disorder due to deficiency of the intra-lysosomal enzyme acid sphingomyelinase (ASM) resulting in intracellular accumulation of sphingomyelin. We evaluated a tandem mass spectrometry (MS/MS) method to analyze ASM activity in dried blood spots (DBS) that may be suitable for laboratory diagnosis of NP including high throughput screening of at-risk populations and potentially for newborn screening. METHODS: ASM activity was measured in 3.2 mm punches from DBS. The eluate was incubated with the ASM substrate (N-Hexanoyl-D-erythro-sphingosylphosphorylcholine [C6-sphingomyelin (C29H59N2O6P)]) and an internal standard (N-butyroyl-D-erythro-sphingosine [C4-ceramide (C22H43NO3)]). ASM product and IS were analyzed using MS/MS in multiple reaction monitoring mode for transitions m/z 370.6>264.3 (ASM internal standard) and m/z 398.6>264.3 (ASM product). RESULTS: ASM activities were stable for up to 2 months at or below 4degrees C. Position of the punch in the DBS and/or hematocrit of the DBS had a limited effect on ASM activities. Both intra- and inter-assay variability were below 10%. There was no carry-over. The median ASM activity in 2,085 newborn infants was 9.5 micromol/h/L (mean 10.6) with a SD of 5.06 micromol/h/L. Six of 2,085 (0.3%) infants were found to have ASM activities below the cut-off of 2.5 micromol/h/L. ASM activities were below the cut-off level in all 10 previously diagnosed cases with NP (range: 0.16 to 2.08 micromol/h/L). CONCLUSIONS: This MS/MS method for the measurement of ASM activity in DBS is robust and suitable for laboratory diagnosis of NP.
*Dried Blood Spot Testing
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Hematocrit
;
Humans
;
Infant, Newborn
;
Reference Standards
;
Sphingomyelin Phosphodiesterase/*analysis/standards
;
Sphingomyelins/metabolism
;
Substrate Specificity
;
*Tandem Mass Spectrometry/standards