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1.
J Drugs Dermatol ; 22(1): 65-73, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-36607758

RESUMO

BACKGROUND: Diabetes mellitus (DM) is a common disease. Seventy percent of patients present with a cutaneous complication, including xerosis. Ceramides-containing (CER) skincare promotes a healthy skin barrier. This international, multicenter, open-label cohort study evaluated twice-daily application for 1 month of CER-containing cleanser and moisturizing cream to improve DM-related xerosis. METHODS: Patients between 18 and 75 years with DM-related xerosis at baseline were eligible. Study visits were on days -30 to 0 (screening), day 0 (baseline), and week 4 (end of study). Evaluations included the Global Aesthetic Improvement Scale (GAIS) and the physician and subject-scored Dry Skin Classification Scale (DSCS). Subject-scored measures of quality of life (QoL) and satisfaction scale with treatment outcomes and product features took place at the end of the study. Tolerance was assessed by monitoring adverse events (AEs). RESULTS: N = 528 subjects from 19 countries completed treatment, the majority having DM type 2 (82.6%). N = 519 (98.3%) met the primary endpoint criteria (GAIS). The CER-containing skincare regimen resulted in statistically significant improvements from baseline (P<0.001) in all parameters of the physician and subject DSCS scores. Patients reported QoL significantly improved by week 4 (P<0.001). At the end of the study, 99.6% (525) of subjects were satisfied with skincare outcomes and product features (99.4% [524]). No product-related AEs were reported during the study. CONCLUSION: CER-containing cleanser and moisturizer were associated with statistically significant improvements in DM-associated xerosis, physician and subject scored severity, patient satisfaction, and improved QoL. The skincare regimen was well tolerated. J Drugs Dermatol. 2023;22(1):65-73. doi:10.36849/JDD.7168.


Assuntos
Diabetes Mellitus , Qualidade de Vida , Humanos , Estudos de Coortes , Ceramidas , Pele , Resultado do Tratamento , Creme para a Pele/uso terapêutico
2.
Chem Pharm Bull (Tokyo) ; 71(1): 31-40, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36596510

RESUMO

Intercellular lipids fill the interstices of corneocytes and serve a barrier function. The amount of transdermal water evaporation varies depending on the packing structure of intercellular lipids, as this structure is important for maintaining barrier efficacy. This packing structure consists of a mixture of crystals (orthorhombic and hexagonal) and liquid crystals (fluid phase), and the proportion of these phases is thought to affect barrier function. However, there have been no methods to visualize the actual distribution of the domains formed by packing structure in intercellular lipids. In this study, the planar distribution of intercellular lipid structures was determined using focal plane array (FPA)-based Fourier transform (FT) IR imaging analysis of stratum corneum cell units obtained by grid stripping. The lipid composition of ceramides was revealed by electrospray ionization tandem mass spectrometry (ESI-MS/MS)-based shotgun lipidomics. The distribution of domains formed by packing structures and the lipid composition of ceramides was compared in skin with high- or low-transepidermal water loss (TEWL). The orthorhombic proportion was lower in high-TEWL skin than in low-TEWL skin. ESI-MS/MS-based shotgun lipidomics analysis showed that the alpha-hydroxyceramide content in the low- and high-TEWL groups differed regarding the distribution of fatty acid chain lengths. The evaluation of stratum corneum cell units using FPA-based FTIR imaging is an innovative technology that can visualize the distribution of domains formed by intercellular lipid-packing structures. Increased proportions of alpha-hydroxyceramide subclasses such as alpha-hydroxy-sphingosine ceramide and alpha-hydroxy-phytosphingosine ceramide were associated with a reduced proportion of the orthorhombic packing structure domain.


Assuntos
Epiderme , Espectrometria de Massas em Tandem , Epiderme/química , Pele/química , Ácidos Graxos , Ceramidas/química , Água/química
3.
J Wound Ostomy Continence Nurs ; 50(1): 31-38, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36640162

RESUMO

PURPOSE: The aim of this study was to determine whether a difference exists in the financial impact of the use of a 2-piece ceramide-infused skin barrier (CIB) versus standard of care barrier (SOC) in Ontario and Alberta using a cost-effectiveness model over a 1-year period for people with a fecal or urinary ostomy. DESIGN: A cost-effectiveness model adapted from a previously published work. SUBJECTS AND SETTING: The model was populated with data inputs from a hypothetical cohort of 1000 individuals in Ontario and 4000 in Alberta. Model results were assessed for robustness via the use of deterministic and probabilistic sensitivity analyses. The provinces of Ontario and Alberta were chosen because cost data were readily accessible. The combined population of these provinces accounts for 50% of Canada's population. RESULTS: An expected cost savings of Can$443.13 (US $322.60) and Can$243.84 (US $177.52) per user for the hypothetical cohort of 1000 individuals in Ontario and 4000 in Alberta per year was obtained for those using a CIB versus a non-infused skin barrier in Ontario and Alberta, respectively. The incremental cost effectiveness ratio (ICER) of CIB to SOC per peristomal skin complication (PSC) avoided and per quality-adjusted life day (QALD) gained was approximately Can$2702 (US $1967)/PSC and Can$1266 (US $922)/QALD for Ontario and approximately Can$1487 (US $1083)/PSC and Can$697 (US $507)/QALD for Alberta. Analysis indicated CIBs remained cost-effective across all sensitivity analyses performed. CONCLUSIONS: Finding suggest that a CIB is cost-effective when compared to a barrier not infused with ceramide when applied to persons with an ostomy and residing in the provinces of Alberta and Ontario.


Assuntos
Ceramidas , Estomia , Humanos , Análise Custo-Benefício , Ceramidas/uso terapêutico , Canadá , Avaliação de Resultados em Cuidados de Saúde
4.
Arch Biochem Biophys ; 735: 109520, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36646267

RESUMO

Lipid regulation of ion channels is a fundamental mechanism in physiological processes as of neurotransmitter release and hormone secretion. Ceramide is a bioactive lipid proposed as a regulator of several voltage-gated ion channels including potassium channels (Kv). It is generated either de novo or by sphingomyelin (SM) hydrolysis in membranes of mammalian cells. In pancreatic ß-cells, ceramide is the main sphingolipid associated with lipotoxicity and likely involved in cell dysfunction. Despite of the wealth of information regarding regulation of potassium channels by ceramides, the regulation of Kv channels by accumulated ceramide in native pancreatic ß-cells has not been investigated. To do so, we used either the C2-ceramide, a cell-permeable short-chain analogue, or a sphingomyelinase (SMase C), a hydrolase causing ceramide to elevate from an endogenous production, in pancreatic ß-cells of rat. C2-ceramide markedly accelerates steady-state current inactivation according to kinetic changes in the channel machinery. Interestingly, only C2-ceramide accelerates current inactivation while SMase C decreases both, peak-current and step-current amplitude supporting differential effects of ceramide derivatives. A specific inhibitor of the Kv2.1 channel (GxTX-1E), readily inhibits a fraction of the Kv channel current while no further inhibition by C2-ceramide superfusion can be observed supporting Kv2.1 channel involvement in the ceramide inhibition. Thus, intramembrane ceramide accumulation, as a lipidic metabolite released under cell-stress conditions, may alter pancreatic ß-cell repolarization and secretion. These results may provide a new insight regarding lipid-protein regulation and advance our understanding in ceramide actions on Kv channels in pancreatic ß-cells.


Assuntos
Células Secretoras de Insulina , Canais de Potássio , Ratos , Animais , Canais de Potássio/metabolismo , Ceramidas/metabolismo , Ceramidas/farmacologia , Células Secretoras de Insulina/metabolismo , Canais Iônicos/metabolismo , Mamíferos/metabolismo
5.
Biomolecules ; 13(1)2023 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-36671552

RESUMO

Cardiovascular diseases (CVDs) are the leading cause of death and illness in Europe and worldwide, responsible for a staggering 47% of deaths in Europe. Over the past few years, there has been increasing evidence pointing to bioactive sphingolipids as drivers of CVDs. Among them, most studies place emphasis on the cardiovascular effect of ceramides and sphingosine-1-phosphate (S1P), reporting correlation between their aberrant expression and CVD risk factors. In experimental in vivo models, pharmacological inhibition of de novo ceramide synthesis averts the development of diabetes, atherosclerosis, hypertension and heart failure. In humans, levels of circulating sphingolipids have been suggested as prognostic indicators for a broad spectrum of diseases. This article provides a comprehensive review of sphingolipids' contribution to cardiovascular, cerebrovascular and metabolic diseases, focusing on the latest experimental and clinical findings. Cumulatively, these studies indicate that monitoring sphingolipid level alterations could allow for better assessment of cardiovascular disease progression and/or severity, and also suggest them as a potential target for future therapeutic intervention. Some approaches may include the down-regulation of specific sphingolipid species levels in the circulation, by inhibiting critical enzymes that catalyze ceramide metabolism, such as ceramidases, sphingomyelinases and sphingosine kinases. Therefore, manipulation of the sphingolipid pathway may be a promising strategy for the treatment of cardio- and cerebrovascular diseases.


Assuntos
Ceramidas , Esfingolipídeos , Humanos , Esfingolipídeos/metabolismo , Ceramidas/metabolismo , Esfingosina/metabolismo , Pulmão/metabolismo , Ceramidases/metabolismo , Lisofosfolipídeos/metabolismo , Biomarcadores
6.
Int J Biol Sci ; 19(1): 311-330, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36594091

RESUMO

Non-alcoholic fatty liver disease (NAFLD), as one of the main causes of chronic liver disease worldwide, encompasses a spectrum of liver conditions that are not caused by other etiology, such as overt alcohol consumption, from simple steatosis to more aggressive non-alcoholic steatohepatitis (NASH) that involves liver inflammation and fibrosis, and to the lethal cirrhosis that may result in liver cancer and liver failure. The molecular mechanisms governing the transition from steatosis to NASH remain not fully understood, but the hepatic lipidome is extensively altered in the setting of steatosis and steatohepatitis, which also correlate with disease progression. With the tremendous advancement in the field of lipidomics in last two decades, a better understanding of the specific role of sphingolipids in fatty liver disease has taken shape. Among the numerous lipid subtypes that accumulate, ceramides are particularly impactful. On the one hand, excessive ceramides deposition in the liver cause hepatic steatosis. On the other hand, ceramides as lipotoxic lipid have significant effects on hepatic inflammation, apoptosis and insulin resistance that contribute to NAFLD. In this review, we summarize and evaluate current understanding of the multiple roles of ceramides in the onset of fatty liver disease and the pathogenic mechanisms underlying their effects, and we also discuss recent advances and challenges in pharmacological interventions targeting ceramide metabolism for the treatment of NAFLD.


Assuntos
Hepatopatia Gordurosa não Alcoólica , Humanos , Hepatopatia Gordurosa não Alcoólica/metabolismo , Ceramidas/metabolismo , Ceramidas/farmacologia , Ceramidas/uso terapêutico , Fosfatos/metabolismo , Fígado/metabolismo , Esfingolipídeos/metabolismo
7.
Int J Mol Sci ; 24(2)2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36674974

RESUMO

Sphingolipids are crucial molecules of the mammalian epidermis. The formation of skin-specific ceramides contributes to the formation of lipid lamellae, which are important for the protection of the epidermis from excessive water loss and protect the skin from the invasion of pathogens and the penetration of xenobiotics. In addition to being structural constituents of the epidermal layer, sphingolipids are also key signaling molecules that participate in the regulation of epidermal cells and the immune cells of the skin. While the importance of ceramides with regard to the proliferation and differentiation of skin cells has been known for a long time, it has emerged in recent years that the sphingolipid sphingosine 1-phosphate (S1P) is also involved in processes such as the proliferation and differentiation of keratinocytes. In addition, the immunomodulatory role of this sphingolipid species is becoming increasingly apparent. This is significant as S1P mediates a variety of its actions via G-protein coupled receptors. It is, therefore, not surprising that dysregulation in the signaling pathways of S1P is involved in the pathophysiological conditions of skin diseases. In the present review, the importance of S1P in skin cells, as well as the immune cells of the skin, is elaborated. In particular, the role of the molecule in inflammatory skin diseases will be discussed. This is important because interfering with S1P signaling pathways may represent an innovative option for the treatment of inflammatory skin diseases.


Assuntos
Dermatite , Dermatopatias , Animais , Esfingosina/metabolismo , Esfingolipídeos/metabolismo , Lisofosfolipídeos/metabolismo , Ceramidas/metabolismo , Dermatopatias/tratamento farmacológico , Dermatopatias/metabolismo , Mamíferos/metabolismo
8.
Skin Res Technol ; 29(1): e13276, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36704885

RESUMO

BACKGROUND: Physiological skin properties of neonates and infants change drastically after birth and are implicated in the onset of atopic dermatitis and other diseases. Studies have measured physiological skin properties in infants; however, how these properties change over time remains unclear. No reports have measured ceramide in the stratum corneum of infants using confocal Raman spectroscopy; hence, we used it to measure the physiological properties of the skin, including ceramide, in infants. MATERIALS AND METHODS: The water content and other factors in the skin of infants aged 0, 1, and 6 months were measured. All measurements were performed five times indoors at 22 ± 2°C and 50% ± 10% relative humidity in the middle of the calf at 4-µm distances, and their mean was calculated. RESULTS: The water content of the area between the skin surface and superficial layers was the lowest in newborns as compared with other ages, and the deeper the skin layer, the higher the water content. The stratum corneum, evaluated using confocal Raman spectroscopy, was the thickest in newborns and gradually thinned with age. Its water content was the lowest in newborns. The levels of natural moisturizing factor, ceramide, and cholesterol were higher in newborns and tended to decrease with age. CONCLUSION: This report is the first to evaluate ceramide in the stratum corneum of infants using confocal Raman spectroscopy and could help in conducting subsequent longitudinal measurements of physiological skin properties in neonates and infants.


Assuntos
Análise Espectral Raman , Humanos , Lactente , Recém-Nascido , Projetos Piloto , Análise Espectral Raman/métodos , Epiderme , Pele/diagnóstico por imagem , Pele/química , Água/análise , Ceramidas/análise
9.
Neurobiol Dis ; 177: 106009, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36689912

RESUMO

Heavy alcohol consumption causes neuronal cell death and cognitive impairment. Neuronal cell death induced by ethanol may result from increased production of the sphingolipid metabolite ceramide. However, the molecular mechanisms of neuronal cell death caused by ethanol-induced ceramide production have not been elucidated. Therefore, we investigated the mechanism through which ethanol-induced ceramide production causes neuronal cell apoptosis using human induced-pluripotent stem cell-derived neurons and SH-SY5Y cells and identified the effects of ceramide on memory deficits in C57BL/6 mice. First, we found that ethanol-induced ceramide production was decreased by inhibition of the de novo synthesis pathway, mediated by serine palmitoyltransferase (SPT). The associated alterations of the molecules related to the ceramide pathway suggest that the elevated level of ceramide activated protein phosphatase 1 (PP1), which inhibited the nuclear translocation of serine/arginine-rich splicing factor 1 (SRSF1). This led to aberrant splicing of myeloid cell leukemia 1 (MCL-1) pre-mRNA, which upregulated MCL-1S expression. Our results demonstrated that the interaction of MCL-1S with the inositol 1, 4, 5-trisphosphate receptor (IP3R) increases calcium release from the endoplasmic reticulum (ER) and then activated ER-bound inverted formin 2 (INF2). In addition, we discovered that F-actin polymerization through INF2 activation promoted ER-mitochondria contacts, which induced mitochondrial calcium influx and mitochondrial reactive oxygen species (mtROS) production. Markedly, MCL-1S silencing decreased mitochondria-associated ER membrane (MAM) formation and prevented mitochondrial calcium influx and mtROS accumulation, by inhibiting INF2-dependent actin polymerization interacting with mitochondria. Furthermore, the inhibition of ceramide production in ethanol-fed mice reduced MCL-1S expression, neuronal cell death, and cognitive impairment. In conclusion, we suggest that ethanol-induced ceramide production may lead to mitochondrial calcium overload through MCL-1S-mediated INF2 activation-dependent MAM formation, which promotes neuronal apoptosis.


Assuntos
Ceramidas , Neuroblastoma , Humanos , Camundongos , Animais , Ceramidas/metabolismo , Etanol/farmacologia , Cálcio/metabolismo , Camundongos Endogâmicos C57BL , Neuroblastoma/metabolismo , Apoptose , Mitocôndrias/metabolismo , Retículo Endoplasmático/metabolismo , Fatores de Processamento de Serina-Arginina
10.
Nutrients ; 15(1)2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36615886

RESUMO

Autoimmune rheumatic diseases (AIRDs) constitute a set of connective tissue disorders and dysfunctions with akin clinical manifestations and autoantibody responses. AIRD treatment is based on a comprehensive approach, with the primary aim being achieving and attaining disease remission, through the control of inflammation. AIRD therapies have a low target specificity, and this usually propels metabolic disturbances, dyslipidemias and increased cardiovascular risk. Ceramides are implicated in inflammation through several different pathways, many of which sometimes intersect. They serve as signaling molecules for apoptosis, altering immune response and driving endothelial dysfunction and as regulators in the production of other molecules, including sphingosine 1-phosphate (S1P) and ceramide 1-phosphate (C1P). With lipid metabolism being severely altered in AIRD pathology, several studies show that the concentration and variety of ceramides in human tissues is altered in patients with rheumatic diseases compared to controls. As a result, many in vitro and some in vivo (animal) studies research the potential use of ceramides as therapeutic targets in rheumatoid arthritis (RA), ankylosing spondylitis, systemic lupus erythematosus, fibromyalgia syndrome, primary Sjögren's syndrome, systemic sclerosis, myositis, systemic vasculitis and psoriatic arthritis. Furthermore, the majority of ceramide synthesis is diet-centric and, as a result, dietary interventions may alter ceramide concentrations in the blood and affect health. Subsequently, more recently several clinical trials evaluated the possibility of distinct dietary patterns and nutrients to act as anti-ceramide regimes in humans. With nutrition being an important component of AIRD-related complications, the present review details the evidence regarding ceramide levels in patients with AIRDs, the results of anti-ceramide treatments and discusses the possibility of using medical nutritional therapy as a complementary anti-ceramide treatment in rheumatic disease.


Assuntos
Artrite Psoriásica , Doenças Autoimunes , Doenças Reumáticas , Animais , Humanos , Ceramidas/metabolismo , Doenças Autoimunes/tratamento farmacológico , Inflamação/metabolismo , Dieta , Doenças Reumáticas/tratamento farmacológico
11.
Anal Biochem ; 661: 114982, 2023 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-36375519

RESUMO

BACKGROUND: Ceramide is one type of sphingolipids, is associated with the occurrence of metabolic diseases, including obesity, diabetes, cardiovascular disease, cancer, and nonalcoholic fatty liver disease. Dihydroceramide, the direct precursors of ceramide, which is converted to ceramide with the dihydroceramide desaturase, is recently regarded as involving in various biological processes and metabolic diseases. The liver and gut ceramide levels are interactional in pathophysiological condition, quantifying hepatic and intestinal ceramide levels become indispensable. The aim of this study is to establish a rapid method for the determination of ceramides including dihydroceramides in liver and small intestinal tissues for researching the mechanisms of ceramide related diseases. METHODS: The levels of Cer d18:1/2:0, Cer d18:1/6:0, Cer d18:1/12:0, Cer d18:1/14:0, Cer d18:1/16:0, Cer d18:1/17:0, Cer d18:1/18:0, Cer d18:1/20:0, Cer d18:1/22:0, Cer d18:1/24:1, Cer d18:1/24:0, dHCer d18:0/12:0, dHCer d18:0/14:0, dHCer d18:0/16:0, dHCer d18:0/18:0, dHCer d18:0/24:1 and dHCer d18:0/24:0 in mice liver and small intestine were directly quantified by ultra-high performance liquid chromatography-tandem mass spectrometry after methanol extraction. In detail, liver or small intestine tissues were thoroughly homogenized with methanol. The resultant ceramides were separated on a Waters BEH C18 column using gradient elution within 10 min. Positive electrospray ionization with multiple reaction monitoring was applied to detect. In the end, the levels of ceramides in mice liver and small intestine tissues were quantified by this developed method. RESULTS: The limits of detection and quantification of 11 ceramides and 6 dihydroceramides were 0.01-0.5 ng/mL and 0.02-1 ng/mL, respectively, and all detected ceramides had good linearities (R2 > 0.997). The extraction recoveries of ceramides at three levels were within 82.32%-115.24% in the liver and within 83.21%-118.70% in the small intestine. The relative standard deviations of intra- and inter-day precision were all within 15%. The extracting solutions of the liver and small intestine could be stably stored in the autosampler 24 h at 10 °C, the lyophilized liver and small intestine for ceramides quantification could be stably stored at least 1 week at -80 °C. The ceramides and dihydroceramides in normal mice liver and small intestinal tissues analyzed by the developed method indicated that the detected 9 ceramide and 5 dihydroceramides levels were significantly different, in which Cer d18:1/16:0, Cer d18:1/22:0, Cer d18:1/24:1, Cer d18:1/24:0 and dHCer d18:0/24:1 are the main components in the liver, whereas Cer d18:1/16:0 and dHCer d18:0/16:0 accounts for the majority of proportion in the intestinal tissues. CONCLUSION: A simple and rapid method for the quantification of 11 ceramides and 6 dihydroceramides in the animal tissues was developed and applied. The compositions of ceramides in two tissues suggested that the compositional features should to be considered when exploring the biomarkers or molecular mechanisms.


Assuntos
Doenças Metabólicas , Espectrometria de Massas em Tandem , Camundongos , Animais , Cromatografia Líquida de Alta Pressão , Metanol , Cromatografia Líquida , Ceramidas , Fígado
12.
Methods Mol Biol ; 2610: 1-16, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36534277

RESUMO

Sphingolipids are a critical family of membrane lipids with diverse functions in eukaryotic cells, and a growing body of literature supports that these lipids play essential roles during the lifecycles of viruses. While small molecule inhibitors of sphingolipid synthesis and metabolism are widely used, the advent of CRISPR-based genomic editing techniques allows for nuanced exploration into the manners in which sphingolipids influence various stages of viral infections. Here we describe some of these critical considerations needed in designing studies utilizing genomic editing techniques for manipulating the sphingolipid metabolic pathway, as well as the current body of literature regarding how viruses depend on the products of this pathway. Here, we highlight the ways in which sphingolipids affect viruses as these pathogens interact with and influence their host cell and describe some of the many open questions remaining in the field.


Assuntos
Esfingolipídeos , Viroses , Humanos , Esfingolipídeos/metabolismo , Lipídeos de Membrana , Ceramidas/metabolismo , Esfingosina/metabolismo
13.
Exp Neurol ; 359: 114263, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36336029

RESUMO

BACKGROUND: Septic-associated encephalopathy (SAE) is a critical manifestation of sepsis that leads to long-term cognitive impairment. Interleukin (IL)-17A has been shown to mediate neuronal apoptosis in central nervous system diseases, while oxidative stress has been found to have a detrimental effect in SAE. However, the relationship between IL-17A and oxidative stress in SAE remains unclear. This study aimed to investigate the effects of secukinumab on alleviating cognitive impairment in a rat model of sepsis, as well as examine its underlying molecular mechanism of action. METHODS: A total of 282 male 8-week-old Sprague-Dawley rats were randomly subjected to cecal ligation and puncture (CLP) or sham treatment followed by volume resuscitation immediately after surgery. Secukinumab was administered intranasally 1 h post-CLP. Rats were given the p-ERK activator ceramide C6 intracerebroventricularly (i.c.v) 24 h before CLP surgery. Recombinant rIL-17A was administered i.c.v. at 0 h in naive rats, followed by intraperitoneal injection of the AKT inhibitor GDC0068 1 h post-rIL-17A injection. Clinical scores, body weight, and survival rate were assessed. In addition, immunofluorescence staining, neurobehavioral tests, Nissl staining, and western blotting were performed. Cognitive function was assessed 15-20 days post-CLP using the Morris water maze test. RESULTS: IL-17A and IL-17RA protein expression levels in the rat hippocampus increased and peaked 24 h post-CLP. Furthermore, IL-17RA was found to be expressed in neurons. The survival rate after CLP was 50%. Following CLP, an increased clinical score and significant decrease in body weight were observed. However, treatment with secukinumab led to a decrease in the clinical score of rats 24 h post-CLP. CLP resulted in spatial and memory impairment and anxiety-like behaviors in rats, while secukinumab treatment significantly alleviated cognitive impairment compared to the CLP group (p < 0.05). In addition, oxidative stress and neuronal apoptosis were found to be increased in the CLP group, while secukinumab significantly reduced oxidative stress and neuronal apoptosis in the hippocampus following CLP. Furthermore, secukinumab treatment led to a significant decrease in the protein expression levels of p-AKT, p-ERK1/2, Romo1, and Bax, together with increased Bcl-2 protein expression. Finally, treatment with ceramide C6 and GDC0068 abolished the neuroprotective effects of secukinumab post-CLP. CONCLUSION: Our results demonstrated that secukinumab attenuated oxidative stress and neuronal apoptosis and partially ameliorated cognitive impairment via the IL-17RA/AKT/ERK1/2 pathway in a rat model of sepsis. Thus, secukinumab may be a potential therapeutic strategy for septic patients.


Assuntos
Disfunção Cognitiva , Encefalopatia Associada a Sepse , Sepse , Animais , Ratos , Masculino , Interleucina-17/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos Sprague-Dawley , Sistema de Sinalização das MAP Quinases , Sepse/complicações , Sepse/tratamento farmacológico , Apoptose , Estresse Oxidativo , Disfunção Cognitiva/tratamento farmacológico , Disfunção Cognitiva/etiologia , Ceramidas/farmacologia , Peso Corporal
14.
Methods Mol Biol ; 2613: 111-125, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36587075

RESUMO

Sphingolipids are ubiquitously expressed in eukaryotes and play various functional roles. The key characteristic of sphingolipids is their diversity of molecular species. Sphingomyelin (SM) and glycosphingolipids (GSLs) are the major components of sphingolipids in the plasma membrane, which are composed of ceramide and a polar head-group. SM is the most abundant sphingolipid species in mammalian cells, while GSLs have a wide variety of glycans as head groups. Various fatty acids in ceramide also contribute to the diversity of sphingolipid species. To analyze the cellular function of each sphingolipid species, precise gene manipulation is essential. Recent developments in genome editing technologies have facilitated complete gene disruption in cultured cells. This chapter describes protocols for the construction of various sphingolipid-related gene knockout HeLa cells using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system and for confirmation of changes in their lipid composition using radioisotopes and thin layer chromatography. This sphingolipid-remodeled cell panel is a useful tool for analyzing the cellular functions of sphingolipid species and as a reference for lipid analysis.


Assuntos
Edição de Genes , Esfingolipídeos , Animais , Humanos , Esfingolipídeos/metabolismo , Edição de Genes/métodos , Células HeLa , Ceramidas/metabolismo , Esfingomielinas/metabolismo , Glicoesfingolipídeos , Sistemas CRISPR-Cas , Mamíferos/metabolismo
15.
Methods Mol Biol ; 2613: 89-100, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36587073

RESUMO

Synthetic methodologies for gangliosides have evolved over the past three decades. The strategies for constructing ganglioside skeletons can generally be classified as late-stage ceramide coupling, the glucosyl ceramide cassette strategy, or late-stage sialylation. Using these synthetic strategies, numerous natural gangliosides and their structural analogs, including functional probes, have been synthesized. This chapter describes the synthetic strategies for gangliosides and provides examples of the total synthesis of several gangliosides using each strategy.


Assuntos
Gangliosídeos , Glucosilceramidas , Gangliosídeos/química , Ceramidas/química
16.
Anal Chem ; 95(2): 1176-1183, 2023 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-36574465

RESUMO

Gangliosides are acidic glycosphingolipids, containing ceramide moieties and oligosaccharide chains with one or more sialic acid residue(s) and are highly diverse isomeric structures with distinct biological roles. Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) enables the untargeted spatial analysis of gangliosides, among other biomolecules, directly from tissue sections. Integrating trapped ion mobility spectrometry with MALDI IMS allows for the analysis of isomeric lipid structures in situ. Here, we demonstrate the gas-phase separation and identification of disialoganglioside isomers GD1a and GD1b that differ in the position of a sialic acid residue, in multiple samples, including a standard mixture of both isomers, a biological extract, and directly from thin tissue sections. The unique spatial distributions of GD1a/b (d36:1) and GD1a/b (d38:1) isomers were determined in rat hippocampus and spinal cord tissue sections, demonstrating the ability to structurally characterize and spatially map gangliosides based on both the carbohydrate chain and ceramide moieties.


Assuntos
Gangliosídeos , Ácido N-Acetilneuramínico , Camundongos , Ratos , Animais , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Gangliosídeos/análise , Encéfalo , Ceramidas
17.
Anal Bioanal Chem ; 415(5): 801-808, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36482083

RESUMO

Ceramides are important intermediates in the metabolism of sphingolipids. High-throughput liquid chromatography-mass spectrometry has been used extensively for monitoring the levels of serological ceramides, but is still limited by inadequate coverage or lack of sensitivity. Herein, a rapid, sensitive, and high-throughput isotope dilution liquid chromatography-negative ion electrospray tandem mass spectrometry (IDLC-nESI-MS/MS) method was developed and verified for accurate quantification of 41 ceramides, involving ceramides with C16-20 sphingosine, dihydro-ceramide, and dehydro-ceramide. This method was validated with excellent linearity (R2 > 0.99) and good recovery in the range of 90-110%. Intra- and inter-day imprecision were below 5.57% and 7.83% respectively. The improved high-throughput quantitative method developed in this study may aid in the accurate characterization of ceramides for understanding ceramide biology and application in disease diagnosis.


Assuntos
Ceramidas , Espectrometria de Massas em Tandem , Espectrometria de Massas em Tandem/métodos , Cromatografia Líquida/métodos , Ceramidas/análise , Esfingolipídeos , Isótopos , Cromatografia Líquida de Alta Pressão/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos
18.
J Enzyme Inhib Med Chem ; 38(1): 343-348, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36519337

RESUMO

Ceramide has a key role in the regulation of cellular senescence and apoptosis. As Ceramide levels are lowered by the action of acid ceramidase (AC), abnormally expressed in various cancers, the identification of AC inhibitors has attracted increasing interest. However, this finding has been mainly hampered by the lack of formats suitable for the screening of large libraries. We have overcome this drawback by adapting a fluorogenic assay to a 384-well plate format. The performance of this optimised platform has been proven by the screening a library of 4100 compounds. Our results show that the miniaturised platform is well suited for screening purposes and it led to the identification of several hits, that belong to different chemical classes and display potency ranges of 2-25 µM. The inhibitors also show selectivity over neutral ceramidase and retain activity in cells and can therefore serve as a basis for further chemical optimisation.


Assuntos
Ceramidase Ácida , Neoplasias , Humanos , Ceramidase Ácida/antagonistas & inibidores , Apoptose , Ceramidas/química , Bibliotecas de Moléculas Pequenas
19.
Chem Biodivers ; 20(1): e202200678, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36480444

RESUMO

Purification through repeated column chromatography over silica gel and Sephadex LH-20 of the ethanol extract of the stems of Cissus aralioides (Baker) Planch. resulted in the isolation of a new ceramide, aralioidamide A (1), along with five known compounds (2-6). Their structures were determined by the extensive analyses of their spectroscopic (1D and 2D NMR) and spectrometric data, and comparison with those reported in the literature. Aralioidamide A (1) displayed weak antibacterial activity (MIC=256 µg/mL) against Bacillus subtilis, Staphylococcus aureus and Shigella flexneri and was inactive (MIC>256 µg/mL) against the tested fungi.


Assuntos
Cissus , Vitaceae , Cissus/química , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Ceramidas/farmacologia , Antibacterianos/química , Testes de Sensibilidade Microbiana
20.
Clin Immunol ; 246: 109212, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36563946

RESUMO

Psoriasis is a chronic immune-mediated inflammatory disease. Lipids play an important role in regulating the inflammatory response. However, the alteration of lipids involved in psoriasis particular in skin lesions remain unclear. Here, we performed the lipidomics to investigate lipid profiling in the skin lesions of the imiquimod-induced psoriasis-like dermatitis and psoriasis patients. The findings showed that ceramides phosphate (CerP) and ceramides were enriched in psoriatic lesions compared with controls from both psoriasis patients and psoriasis-like mouse model. Psoriasis patients were classified into two subtypes, the CC1 and CC2, by consensus clustering of these lipid signatures. The CC1 was characterized by the higher levels of CerP, uric acid, and more severe psoriasis, compared with CC2 subtype. Interestingly, ceramide-1-phosphate (C1P), dramatically enriched in CC1 subtype, facilitated imiquimod-induced psoriasis-like inflammatory responses. Mechanistically, C1P induced the expression of inflammatory factors and activated DNA replication and cell cycle signaling pathways in the primary keratinocytes. Inhibiting the production of C1P with ceramide kinase inhibitor effectively alleviated the imiquimod-induced psoriasis-like inflammation. Taken together, we described the landscape of lipids alteration and established lipids classification based on pattern of abundance of lipids in psoriatic skin lesions. Suppression of C1P pathway is a novel potential strategy for psoriasis treatment.


Assuntos
Lipidômica , Psoríase , Animais , Camundongos , Imiquimode/farmacologia , Pele/patologia , Psoríase/tratamento farmacológico , Queratinócitos , Inflamação/patologia , Ceramidas/efeitos adversos , Lipídeos/efeitos adversos , Modelos Animais de Doenças , Camundongos Endogâmicos BALB C
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