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The relationship among amyloid-βdeposition,sphingomyelin level,and the expression and function of P-glycoprotein in Alzheimer’s disease pathological process

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Neural Regeneration Research 2023年第6期18卷 1300-1307页

作者： Zi-Kang Xing Li-Sha Du Xin Fang Heng Liang Sheng-Nan Zhang Lei Shi Chun-Xiang Kuang Tian-Xiong Han Qing Yang State Key Laboratory of Genetic Engineering School of Life SciencesShanghai Engineering Research Center of Industrial MicroorganismsFudan UniversityShanghaiChina Shanghai Key Lab of Chemical Assessment and Sustainability School of Chemical Science and EngineeringTongji UniversityShanghaiChina Department of Traditional Chinese Medicine Tenth People’s Hospital of Tongji UniversityShanghaiChina

In Alzheimer’s disease,the transporter P-glycoprotein is responsible for the clearance of amyloid-βin the brain.Amyloid-βcorrelates with the sphingomyelin metabolism,and sphingomyelin participates in the regulation of P-glycoprotein.The amyloid cascade hypothesis describes amyloid-βas the central cause of Alzheimer’s disease neuropathology.Better understanding of the change of P-glycoprotein and sphingomyelin along with amyloid-βand their potential association in the pathological process of Alzheimer’s disease is critical.Herein,we found that the expression of P-glycoprotein in APP/PS1 mice tended to increase with age and was significantly higher at 9 and 12 months of age than that in wild-type mice at comparable age.The functionality of P-glycoprotein of APP/PS1 mice did not change with age but was significantly lower than that of wild-type mice at 12 months of age.Decreased sphingomyelin levels,increased ceramide levels,and the increased expression and activity of neutral sphingomyelinase 1 were observed in APP/PS1 mice at 9 and 12 months of age compared with the levels in wild-type mice.Similar results were observed in the Alzheimer’s disease mouse model induced by intracerebroventricular injection of amyloid-β1-42 and human cerebral microvascular endothelial cells treated with amyloid-β1-42.In human cerebral microvascular endothelial cells,neutral sphingomyelinase 1 inhibitor interfered with the changes of sphingomyelin metabolism and P-glycoprotein expression and functionality caused by amyloid-β1-42 treatment.Neutral sphingomyelinase 1 regulated the expression and functionality of P-glycoprotein and the levels of sphingomyelin and ceramide.Together,these findings indicate that neutral sphingomyelinase 1 regulates the expression and function of P-glycoprotein via the sphingomyelin/ceramide pathway.These studies may serve as new pursuits for the development of anti-Alzheimer’s disease drugs.

关键词： Alzheimer’s disease amyloid-β APP/PS1 mice ceramide ezrin-radixin-moesin human cerebral microvascular endothelial cells neutral sphingomyelinase 1 P-glycoprotein sphingomyelin synthase sphingomyelin

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A Lipidomic Study of the Effects of N-methyl-N′-nitro-N-nitrosoguanidine on sphingomyelin Metabolism

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Acta Biochimica et Biophysica Sinica 2005年第8期37卷 515-524页

作者： Yun HUANG~(&#) Jing SHEN~# Ting WANG Yan-Ke YU Fanqing E CHEN~1 Jun YANG~* Department of Pathology and Pathophysiology,Center for Environmental Genomics Zhejiang University School of Medicine,Hangzhou 310031,China ~1 Molecular Biology Branch.Life Science Division,Lawrence Berkeley National Laboratory,University of California at Berkeley,Berkeley,CA 94720,USA Department of Chemistry,Georgia State University.Atlanta,GA 30303.USA Department of Pathology and Pathophysiology Center for Environmental Genomics Zhejiang University School of MedicineHangzhou 310031 China Molecular Biology Branch Life Science Division Lawrence Berkeley National Laboratory University of California at BerkeleyBerkeley CA 94720 USA

Systems biology is a new and rapidly developing research area in which,by quantitativelydescribing the interaction among all the individual components of a cell,a systems-level understanding of abiological response can be achieved.Therefore,it requires high-throughput measurement technologies forbiological molecules,such as genomic and proteomic approaches for DNA/RNA and protein,respectively.Recently,a new concept,lipidomics,which utilizes the mass spectrometry(MS)method for lipid analysis,has been proposed.Using this lipidomic approach,the effects of N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)on sphingomyelin metabolism,a major class of sphingolipids,were evaluated.sphingomyelin moleculeswere extracted from cells and analyzed by matrix-assisted laser desorption ionization-time of flight MS.Itwas found that MNNG induced profound changes in sphingomyelin metabolism,including the appearance ofsome new sphingomyelin species and the disappearance of some others,and the concentrations of severalsphmgomyelin species also changed.This was accompanied by the redistribution of acid sphingomyelinase(ASM),a key player in sphingomyelin metabolism.On the other hand,imipramine,an inhibitor of ASM,caused the accumulation of sphingomyelin.It also prevented some of the effects of MNNG,as well as theredistribution of ASM.Taken together,these data suggested that the lipidomic approach is highly effectivefor the systematic analysis of cellular lipids metabolism.

关键词： lipidomics mass spectrometry ceramide sphingomyelin acid sphingomyelinase

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Endogenous ceramide phosphoethanolamine modulates circadian rhythm via neural-glial coupling in Drosophila

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National Science Review 2022年第12期9卷 104-123页

作者： Xiupeng Chen Jie Li Zhongbao Gao Yang Yang Wenqing Kuang Yue Dong Gek Huey Chua Xiahe Huang Binhua Jiang He Tian Yingchun Wang Xun Huang Yan Li Sin Man Lam Guanghou Shui State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental Biology Chinese Academy of Sciences University of Chinese Academy of Sciences State Key Laboratory of Brain and Cognitive Science Institute of Biophysics Chinese Academy of Sciences Lipid ALL Technologies Company Limited

While endogenous lipids are known to exhibit rhythmic oscillations,less is known about how specific lipids modulate circadian behavior.Through a series of loss-of-function and gain-of-function experiments on cer amide phosphoethanolamine(CPE) synthase of Drosophila,we demonstrated that pan-glial-specific deficiency in membrane CPE,the structural analog of mammalian sphingomyelin(SM),leads to arrhythmic locomotor behavior and shortens lifespan,while the reverse is true for increasing CPE.Comparative proteomics uncovered dysregulated synaptic glutamate utilization and transport in CPE-deficient flies.An extensive genetic screen was conducted to verify the role of differentially expressed proteins in circadian regulation.Arrhythmic locomotion under cpes1mutant background was rescued only by restoring endogenous CPE or SM through expressing their respective synthases.Our results underscore the essential role of CPE in maintaining synaptic glutamate homeostasis and modulating circadian behavior in Drosophila.The findings suggest that region-specific elevations of functional membrane lipids can benefit circadian regulation.

关键词： ceramide phosphoethanolamine sphingomyelin circadian longevity glutamate homeostasis

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A novel mutation in ROR2 led to the loss of function of ROR2 and inhibited the osteogenic differentiation capability of bone marrow mesenchymal stem cells(BMSCs)

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BIOCELL 2023年第7期47卷 1561-1569页

作者： WENQI CHEN XIAOYANG CHU YANG ZENG YOUSHENG YAN YIPENG WANG DONGLAN SUN DONGLIANG ZHANG JING ZHANG KAI YANG Prenatal Diagnosis Center Shijiazhuang Obstetrics and Gynecology HospitalKey Laboratory of Maternal and Fetal Medicine of Hebei ProvinceShijiazhuang050011China Department of Stomatology Fifth Medical Center of Chinese PLA General HospitalBeijing100039China Institute of Hematology Fifth Medical Center of Chinese PLA General HospitalBeijing100039China Prenatal Diagnosis Center Beijing Obstetrics and Gynecology HospitalBeijing Maternal and Child Health Care HospitalCapital Medical UniversityBeijing100026China Department of Orthodontics Beijing Stomatological HospitalCapital Medical University School of StomatologyCapital Medical UniversityBeijing100050China

Receptor tyrosine kinase-like orphan receptor 2(ROR2)has a vital role in osteogenesis.However,the mechanism underlying the regulation of ROR2 in osteogenic differentiation is still poorly comprehended.A previous study by our research group showed that a novel compound heterozygous ROR2 variation accounted for the autosomal recessive Robinow syndrome(ARRS).This study attempted to explore the impact of the ROR2:c.904C>T variant specifically on the osteogenic differentiation of BMSCs.Methods:Coimmunoprecipitation(CoIP)-western blotting was carried out to identify the interaction between ROR2 and Wnt5a.Double-immunofluorescence staining was used for determining the expressions and co-localization of ROR2 and Wnt5a in bone marrow mesenchymal stem cells(BMSCs).Western blot(WB)analysis and quantitative reverse transcription polymerase chain reaction(RT-qPCR)were conducted to identify the expression levels of ROR2 in the BMSCs transfected with LV-shROR2 or LV-ROR2-c.904C>T.The alkaline phosphatase(ALP)activity was detected,and Alizarin Red S staining was done for evaluating the osteogenic differentiation of BMSCs.RT-qPCR was employed to identify the expression of the sphingomyelin synthase 1(SMS1)mRNA in the BMSCs transfected with LV-shROR2 or LV-ROR2-c.904C>T and the mRNA expression levels of Runt-related transcription factor 2(RUNX2),osteocalcin(OCN),and osteopontin(OPN).WB was performed to confirm the protein expressions of extracellular regulated protein kinases1(ERK),P-ERK,Smad family member1/5/8(Smad1/5/8),P-Smad1/5/8,P-P38,P38,RUNX2,OCN,and OPN in the BMSCs transfected with LV-shROR2/LV-ROR2-c.904C>T and sphingomyelin(SM).Results:The ROR2:c.904C>T mutant altered the subcellular localization of the ROR2 protein,which caused an impaired interaction between ROR2 and Wnt5a.The depletion of ROR2 restricted the osteogenic differentiation capability of BMSCs and downregulated the expression of SMS1.SM treatment could reverse the inhibition of osteoblastic differentiation in ROR2-depleted BMSCs.Conclusion:The findings of

关键词： Bone marrow mesenchymal stem cells ROR2 Wnt5a Osteogenic differentiation sphingomyelin

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Ceramide is involved in alcohol-induced neural proliferation

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Neural Regeneration Research 2013年第23期8卷 2178-2189页

作者： Zhixin Wang Tongxing Deng Jiexin Deng Jinbo Deng Xiaoqun Gao Yuanyuan Shi Bin Liu Zhanyou Ma Haixiao Jin Institute of Neurobiology Henan University Department of Anatomy Basic Medical CollegeZhengzhou University Department of Anatomy Luohe Medical College

Prenatal alcohol exposure, especially during early pregnancy, can lead to fetal alcohol syndrome. The pharmacological and toxicological mechanisms of ethanol are related to the effects of ceramide In this study, we established an alcohol exposure model in wild-type mice and in knockout mice for the key enzyme involved in ceramide metabolism, sphingomyelin synthase 2. This model received daily intragastric administration of 25% ethanol, and pups were used at postnatal days 0, 7, 14, 30 for experiments. Serology and immunofluorescence staining found that ethanol exposure dose-dependently reduced blood sphingomyelin levels in two genotypes of pups, and increased neural cell proliferation and the number of new neurons in the hippocampal dentate gyrus. Western blot analysis showed that the relative expression level of protein kinase C e increased in two genotypes of pups after ethanol exposure. Compared with witd-type pups, the expression level of the important activator protein of the ceramide/ceramide-l-phosphate pathway, protein kinase C a, was reduced in the hippocampus of sphingomyelin synthase 2 knockouts. Our findings illustrate that ceramide is involved in alcohol-induced neural proliferation in the hippocampal dentate gyrus of pups after prenatal ethanol exposure, and the mechanism may be associated with increased ex- pression of protein kinase C a activating the ceramide/ceramide-l-phosphate pathway.

关键词： neural regeneration brain injury ceramide neural cells proliferation prenatal alcohol exposure sphingomyelin synthase 2 knockout mice sphingomyelin sphingomyelin synthase cera-mide-l-phosphate fetal alcohol syndrome grants-supported paper neuroregeneration

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Alkaline sphingomyelinase(NPP7) in hepatobiliary diseases: A field that needs to be closely studied

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World Journal of Hepatology 2018年第2期10卷 246-253页

作者： Rui-Dong Duan Gastroenterology and Nutrition Lab Department of Clinical Sciences Lund University

Alkaline sphingomyelinase cleaves phosphocholine from sphingomyelin, platelet-activating factor, lysophosphatidylcholine, and less effectively phosphatidyl-choline. The enzyme shares no structure similarities with acid or neutral sphingomyelinase but belongs to ectonucleotide pyrophosphatase/phosphodiesterase(NPP) family and therefore is also called NPP7 nowadays. The enzyme is expressed in the intestinal mucosa in many species and additionally in human liver. The enzyme in the intestinal tract has been extensively studied but not that in human liver. Studies on intestinal alkaline sphingomyelinase show that it inhibits colonic tumorigenesis and inflammation, hydrolyses dietary sphingomyelin, and stimulates cholesterol absorption. The review aims to summarize the current knowledge on liver alkaline sphingomyelinase in human and strengthen the necessity for close study on this unique human enzyme in hepatobiliary diseases.

关键词： sphingomyelin Alkaline sphingomyelinase Nucleotide pyrophosphatase/phosphodiesterase 7 Autotaxin Platelet-activating factor Cholangiocarcinoma Liver diseases Gallstone

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Analysis of Phospholipids in Crucian Carp(Carassius auratus) Muscle by Offline HPLC-MALDI-TOF MS

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Chemical Research in Chinese Universities 2011年第3期27卷 371-374页

作者： LI Guo-chen WANG Yan-hong WU Ren-an WANG Shi-cheng Inspection and Testing Center of Agro-product Safety and Environment Quality Institute of Applied EcologyChinese Academy of SciencesShenyang 110016P.R.China National Chromatographic Research&Analysis Center Dalian Institute of Chemical PhysicsChinese Academy of SciencesDalian 116023P.R.China

The interest in the analysis of phospholipids(PLs), especially phosphatidylcholine(PC), has been increasing due to the importance of them in biochemistry as well as in industry. A method was reported based on the offline combination of MALDI-TOF MS and normal-phase HPLC for analyzing PLs extracted from crucian carp. Total PLs of crucian carp were extracted and then separated by HPLC before the collected subfractions were analyzed by MALDI-TOF MS. The mass spectra obtained show peaks of H+, Na+ and K+ adducts of PC molecules. It is shown that the prior separation of PLs by HPLC is highly necessary to remove the signal suppressing and to avoid the possible overlapping. With this method, 9 possible PC molecules in crucian carp and the corresponding fatty acid compositions were given from the well-resolved mass spectra.

关键词： Phospholipid Phosphatidylcholine Phosphatidylethanolamine sphingomyelin HPLC MALDI Crucian Carp

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Concluding Step in Cell Restitution Cycle: ER Transport Vesicles with Sphingolipids in the Outer Leaflet of the Membrane Restore Lysosomes

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Advances in Biological Chemistry 2014年第5期4卷 301-321页

作者： Amalia Slomiany Bronislaw L. Slomiany Biomedical and Health Sciences School of Dental Medicine Rutgers The State University of New Jersey Newark USA

Restitution of the cell organelles and the membrane implicates serine palmitoyltransferase (SPT) in signal-specific and selective assembly of the transport vesicles. Here, we reveal that SPT, embedded in the outer leaflet (OL) of endoplasmic reticulum (ER), is engaged in the synthesis of ER transport vesicles that recondition cell organelles, and the inner leaflet (IL) SPT in the restitution of the cell membrane. The OL SPT impacts assembly of sphingomyelinase (SMase)—susceptible ER vesicles but not the SMase-resistant and sphingolipid (SPhL) core—carrying vesicles that refurbish the cell membrane. The investigation of the SPT-initiated differences in the placement of SPhL in vesicular membranes by utilizing ER depleted of OL SPT, allows us to conclude that the restitution of endosomal and lysosomal membranes is achieved with the involvement of OL SPT, whereas the IL SPT is involved in formation of the lipid core for glycosphingolipids (GSL) and sphingomyelin (SM) of the apical and basolateral cell membrane. These findings along with our previously published report (Slomiany and Slomiany, Advances in Biological Chemistry, 2013, 3, 275-287), provide a clear distinction between the processes that renovate cell membrane and its organelles from that of the endocytotic cell debridement, and show that vesicles are navigated to the specific organelles and the cell membrane by the biomembrane constituents programmed in ER.

关键词： ER Transport Vesicles Specificity Serine Palmitoyltransferase sphingomyelin Lysosomes Restitution Cell Debridement Autophagocytes

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Interaction of Ca2+ with sphingomyelin Membrane Studied by High-Resolution Broadband Sum Frequency Vibrational Spectroscopy

Interaction of Ca2+ with Sphingomyelin Membrane Studied by H...

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第十五届全国化学动力学会议

作者：张贞 Rong-juan Feng Yi-yi Li Ming-hua Liu Yuan Guo Beijing National Laboratory for Molecular Sciences CAS Research/Education Center for Excellence in Molecular SciencesInstitute of ChemistryChinese Academy of Sciences University of Chinese Academy of Sciences National Center for Nanoscience and Technology Beijing National Laboratory for Molecular Sciences CAS Key Laboratory of ColloidInterface and Chemical ThermodynamicsInstitute of ChemistryChinese Academy of Sciences

sphingomyelin(SM) is specifically enriched in the plasma membrane of mammalian cells. The functions of the SM molecules play crucial roles in the cellular activities by the interaction between the sphingomyelin and Ca ions. In this report, we investigate the conformational differences between the egg sphingomyelin(ESM) and milk sphingomyelin(MSM) monolayer caused by Ca ions using sub-wavenumber high-resolution broadband sum frequency generation vibrational spectroscopy(HR-BB-SFG-VS). The HR-BB-SFG-VS spectra of one alkyl-chain-deuterated ESM and ESM revealed that the Ca ions do not affect the N-linked saturated fatty acid chain, but they make the sphingosine backbone order. [1] Unlike ESM, Ca ions can induce disordering of the acyl chains in the MSM monolayer. In addition, both ESM and MSM show the large blue shifts of the phosphate group at the Ca Cl2 solution interface, which indicates new cation binding modes for ESM and MSM. The binding causes the phosphate moiety to dehydrate result in the conformation change of the phosphate moiety. From above results, we propose the interactional molecular mechanism between Ca and ESM and MSM molecules: firstly, for both ESM and MSM polar headgroup, Ca ions bind to the phosphate group and subsequently destroy the intramolecular hydrogen bond between the 3-hydroxyl group and the phosphate oxygen, which results in the conformation change of the alky chain. Secondly, in ESM monolayer, Ca results in an ordering change of the sphingosine backbone. The interactions between ESM and Ca ions make the orientation of the methyl group at the end of sphingosine backbone change from pointing downward to pointing upward. But in MSM monolayer, the Ca bonded to the phosphate group to create a strong dipolar fields penetrating into the membrane interior. Such electric fields are large enough to induce more gauche-defects of the CH2 groups closed to the head group. However, the addition of Ca do not affect the conformation of the methyl located at the tail of the alky chain. This results she

关键词： high-resolution broadband sum frequency generation vibrational spectroscopy sphingomyelin conformation change lipid–cation interactions

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

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World Journal of Gastrointestinal Pathophysiology 2016年第2期7卷 235-241页

作者： Nobuya Kurabe Hisaki Igarashi Ippei Ohnishi Shogo Tajima Yusuke Inoue Yoshihiko Takahashi Mitsutoshi Setou Haruhiko Sugimura Department of Tumor Pathology Hamamatsu University School of Medicine Division of Pathology Iwata City Hospital Department of Pathology the University of Tokyo Second Division Department of Internal MedicineHamamatsu University School of Medicine Department of Cell Biology and Anatomy Hamamatsu University School of Medicine

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.

关键词： Imaging mass spectrometry i MScope sphingomyelin Phosphatidylcholine Gastric mucosa

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