Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 63
Filtrar
1.
Small ; 18(9): e2105021, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35088527

RESUMO

Atherosclerosis (AS) is associated with high morbidity and mortality, thus imposing a growing burden on modern society. Herb-derived bicyclol (BIC) is a versatile bioactive compound that can be used to treat AS. However, its efficacy in AS is not yet described. Here, it is shown that BIC normalizes gut microflora dysbiosis induced by a high fat diet in Apoe(-/-) mice. Metagenome-wide association study analysis verifies that the modulation on carbohydrate-active enzymes and short-chain fatty acid generating genes in gut flora is among the mechanisms. The gut healthiness, especially the gut immunity and integrity, is restored by BIC intervention, leading to improved systemic immune cell dynamic and liver functions. Accordingly, the endothelial activation, macrophage infiltration, and cholesterol ester accumulation in the aortic arch are alleviated by BIC to lessen the plaque onset. Moreover, it is proved that the therapeutic effect of BIC on AS is transmissible by fecal microbiota transplantation. The current study, for the first time, demonstrates the antiatherosclerotic effects of BIC and shows that its therapeutic value can at least partially be attributed to its manipulation of gut microbiota.


Assuntos
Aterosclerose , Microbioma Gastrointestinal , Animais , Aterosclerose/tratamento farmacológico , Compostos de Bifenilo/farmacologia , Compostos de Bifenilo/uso terapêutico , Disbiose , Camundongos , Camundongos Endogâmicos C57BL
2.
Signal Transduct Target Ther ; 6(1): 414, 2021 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-34873151

RESUMO

Azvudine (FNC) is a nucleoside analog that inhibits HIV-1 RNA-dependent RNA polymerase (RdRp). Recently, we discovered FNC an agent against SARS-CoV-2, and have taken it into Phase III trial for COVID-19 patients. FNC monophosphate analog inhibited SARS-CoV-2 and HCoV-OC43 coronavirus with an EC50 between 1.2 and 4.3 µM, depending on viruses or cells, and selective index (SI) in 15-83 range. Oral administration of FNC in rats revealed a substantial thymus-homing feature, with FNC triphosphate (the active form) concentrated in the thymus and peripheral blood mononuclear cells (PBMC). Treating SARS-CoV-2 infected rhesus macaques with FNC (0.07 mg/kg, qd, orally) reduced viral load, recuperated the thymus, improved lymphocyte profiles, alleviated inflammation and organ damage, and lessened ground-glass opacities in chest X-ray. Single-cell sequencing suggested the promotion of thymus function by FNC. A randomized, single-arm clinical trial of FNC on compassionate use (n = 31) showed that oral FNC (5 mg, qd) cured all COVID-19 patients, with 100% viral ribonucleic acid negative conversion in 3.29 ± 2.22 days (range: 1-9 days) and 100% hospital discharge rate in 9.00 ± 4.93 days (range: 2-25 days). The side-effect of FNC is minor and transient dizziness and nausea in 16.12% (5/31) patients. Thus, FNC might cure COVID-19 through its anti-SARS-CoV-2 activity concentrated in the thymus, followed by promoted immunity.


Assuntos
Antivirais/administração & dosagem , Azidas/administração & dosagem , COVID-19/tratamento farmacológico , Desoxicitidina/análogos & derivados , SARS-CoV-2/metabolismo , Timo , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Coronavirus Humano OC43/metabolismo , Desoxicitidina/administração & dosagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ratos , Timo/metabolismo , Timo/virologia
3.
Anal Chim Acta ; 1132: 74-82, 2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-32980113

RESUMO

Bile acids (BAs), as crucial endogenous metabolites, are closely related to cholestasis, metabolic disorders, and cancer. To better understand their function and disease pathogenesis, global profiling of BAs is necessary. Here, multidimensional data mining was developed for the discovery and identification of potentially unknown BAs in cholestasis rats. Based on an in-house theoretical BA database and using a newly established liquid chromatography-tandem high-resolution mass spectrometry (LC-HRMS/MS) method, four-dimensional (4D) data including the retention times (RT), abundances, HRMS, and HRMS/MS spectra were acquired and elucidated. And 491 BAs were totally profiled. Then, the relationships between RT with different conjugation types, different positions and configurations of hydroxyl/ketone groups as well as fragmentation rules of hydroxyl, ortho-hydroxyl, ketone, and conjugated groups of BAs were summarized to assist BA identification for the first time. Finally, 292 BAs were assigned with molecular formulas, 201 of which were putatively identified by integrating the 4D data, applying structure-driven relative retention time rules, and a comparison with synthetic BAs. The estimated concentrations of 201 BAs, including 93 reported and 108 newly identified BAs, were quantified by using surrogate standards with similar structure. Among 201 BAs, 38 BAs were detected in both humans and rats for the first time. Our strategy has expanded the scope of BAs and provides a way to identify a class of metabolites. Compared to normal rats, the significantly increased sulfated and glucuronide conjugated BAs in urine and feces from experimentally cholestatic rats may reveal a way to diagnose intrahepatic cholestasis.


Assuntos
Ácidos e Sais Biliares , Mineração de Dados , Sulfatos , Animais , Cromatografia Líquida , Espectrometria de Massas , Ratos
4.
Eur Respir J ; 56(5)2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32513782

RESUMO

Pathological mechanisms of pulmonary arterial hypertension (PAH) remain largely unexplored. Effective treatment of PAH remains a challenge. The aim of this study was to discover the underlying mechanism of PAH through functional metabolomics and to help develop new strategies for prevention and treatment of PAH.Metabolomic profiling of plasma in patients with idiopathic PAH was evaluated through high-performance liquid chromatography mass spectrometry, with spermine identified to be the most significant and validated in another independent cohort. The roles of spermine and spermine synthase were examined in pulmonary arterial smooth muscle cells (PASMCs) and rodent models of pulmonary hypertension.Using targeted metabolomics, plasma spermine levels were found to be higher in patients with idiopathic PAH compared to healthy controls. Spermine administration promoted proliferation and migration of PASMCs and exacerbated vascular remodelling in rodent models of pulmonary hypertension. The spermine-mediated deteriorative effect can be attributed to a corresponding upregulation of its synthase in the pathological process. Inhibition of spermine synthase in vitro suppressed platelet-derived growth factor-BB-mediated proliferation of PASMCs, and in vivo attenuated monocrotaline-mediated pulmonary hypertension in rats.Plasma spermine promotes pulmonary vascular remodelling. Inhibiting spermine synthesis could be a therapeutic strategy for PAH.


Assuntos
Hipertensão Arterial Pulmonar , Animais , Proliferação de Células , Modelos Animais de Doenças , Glicogênio Sintase , Humanos , Miócitos de Músculo Liso , Artéria Pulmonar , Ratos , Espermina , Remodelação Vascular
5.
Anal Chem ; 92(12): 8487-8496, 2020 06 16.
Artigo em Inglês | MEDLINE | ID: mdl-32412732

RESUMO

The profile of cholesteryl esters (CEs) is increasingly used in metabolic disease monitoring due to the roles of CE in regulating the cholesterol level. While electrospray ionization-tandem mass spectrometry is routinely applied for the identification and quantitation of CE, it has a limitation of not being able to provide the location of carbon-carbon double bond (C═C) within unsaturated fatty acyls. In this study, we paired offline 2-acetylpyridine (2-AP) Paternò-Büchi (PB) reaction and reversed-phase liquid chromatography-tandem mass spectrometry to achieve highly sensitive and structural informative CE analysis from complex mixtures. The 2-AP PB reactions of CE standards provided 20-30% conversion but resulted in enhanced ion signal relative to that of intact CE detected as ammonium adduct ions. MS/MS of 2-AP derivatized CE via collision-induced dissociation produced two abundant diagnostic ions for each C═C in a fatty acyl, leading to both sensitive identification and quantitation of C═C location isomers. Twelve saturated and twenty-seven unsaturated CEs were profiled in pooled human plasma; of the latter group, relative quantitation of 6 groups of C═C location isomers was achieved. A dehydrocholesteryl ester, DHE 18:2 (Δ9,12), was confidently differentiated from coexisting compositional isomers: CE 18:3 (Δ9,12,15) and CE 18:3 (Δ6,9,12). The above results represented improved CE coverage at the C═C location level over those reported by gas chromatography MS or acetone PB-MS/MS methods.

6.
Chin Med J (Engl) ; 133(2): 198-204, 2020 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-31880746

RESUMO

BACKGROUND: Previously, dihydroceramide (d18:0/24:0) (dhCer (d18:0/24:0)) was reported to be a potential biomarker for acute-on-chronic liver failure (ACLF) prognosis. In this study, we further explored the role of dhCer (d18:0/24:0) in the progression of ACLF to validate the biomarker using ACLF rat model. METHODS: ACLF rats were sacrificed at 4 and 8 h post-D-galactosamine (D-gal)/lipopolysaccharide (LPS) administration to investigate the liver biochemical markers, prothrombin time and liver histopathology. Change in dhCer and other sphingolipids levels were investigated by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). Rats were treated with N-(4-hydroxyphenyl) retinamide (4-HPR) to examine the mortality rate and its role in improving ACLF. RESULTS: LPS/D-gal administration resulted in significant elevation in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Prothrombin time was prolonged and histopathological examination showed abnormality. HPLC-MS/MS results showed total dhCer levels in ACLF group (64.10 ±â€Š8.90 pmol/100 µL, 64.22 ±â€Š6.78 pmol/100 µL for 4 and 8 h, respectively) were decreased significantly compared with control group (121.61 ±â€Š23.09 pmol/100 µL) (P < 0.05). In particular, dhCer (d18:0/24:0), dhCer (d18:0/20:0), and dhCer (d18:0/22:0) levels were decreased. Treatment with 4-HPR significantly increased the levels of dhCers, including dhCer (d18:0/24:0) compared with ACLF group, for the level of dhCer (d18:0/24:0) in 4-HPR group was 20.10 ±â€Š8.60 pmol/100 µL and the level of dhCer (d18:0/24:0) in ACLF group was 9.74 ±â€Š2.99 pmol/100 µL (P < 0.05). This was associated with reduced mortality rate and prolonged survival time. The ALT and AST in 4-HPR group were significantly decreased compared with ACLF group. The prothrombin time of 4-HPR group (41.49 s) was significantly lower than the prothrombin time of ACLF group (57.96 s) (P < 0.05). 4-HPR also decreased plasma ammonia levels slightly, as the plasma ammonia levels in 4-HPR group and ACLF group were 207.37 ±â€Š60.43, 209.15 ±â€Š60.43 µmol/L, respectively. Further, 4-HPR treatment improved histopathological parameters. CONCLUSIONS: DhCer, especially dhCer (d18:0/24:0), is involved in the progression of ACLF. Increasing the levels of dhCer can reduce the mortality rate of ACLF rats and alleviate liver injury.


Assuntos
Insuficiência Hepática Crônica Agudizada/sangue , Insuficiência Hepática Crônica Agudizada/patologia , Ceramidas/sangue , Ceramidas/metabolismo , Insuficiência Hepática Crônica Agudizada/metabolismo , Alanina Transaminase/sangue , Amônia/sangue , Animais , Aspartato Aminotransferases/sangue , Western Blotting , Progressão da Doença , Masculino , Ratos , Ratos Wistar , Esfingolipídeos/sangue
7.
J Sep Sci ; 43(1): 31-55, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31573133

RESUMO

The formation of DNA adducts by genotoxic agents is an early event in cancer development, and it may lead to gene mutations, thereby initiating tumor development. The measurement of DNA adducts can provide critical information about the genotoxic potential of a chemical and its mechanism of carcinogenesis. In recent decades, liquid chromatography coupled with mass spectrometry has become the most important technique for analyzing DNA adducts. The improvements in resolution achievable with new chromatographic separation techniques coupled with the high specificity and sensitivity and wide dynamic range of new mass spectrometry systems have been used for both qualitative and quantitative analyses of DNA adducts. This review discusses the challenges in qualitative and quantitative analyses of DNA adducts by liquid chromatography coupled with mass spectrometry and highlights recent developments towards overcoming the limitations of liquid chromatography coupled with mass spectrometry methods. The key steps and new solutions, such as sample preparation, mass spectrometry fragmentation, and method validation, are summarized. In addition, the fundamental principles and latest advances in DNA adductomic approaches are reviewed.


Assuntos
Adutos de DNA/análise , Sequência de Bases , Cromatografia Líquida , Humanos , Espectrometria de Massas
8.
Theranostics ; 9(23): 6745-6763, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31660066

RESUMO

RATIONALE: Inducing cancer differentiation is a promising approach to treat cancer. Here, we identified chlorogenic acid (CA), a potential differentiation inducer, for cancer therapy, and elucidated the molecular mechanisms underlying its differentiation-inducing effects on cancer cells. METHODS: Cancer cell differentiation was investigated by measuring malignant behavior, including growth rate, invasion/migration, morphological change, maturation, and ATP production. Gene expression was analyzed by microarray analysis, qRT-PCR, and protein measurement, and molecular biology techniques were employed for mechanistic studies. LC/MS analysis was the method of choice for chemical detection. Finally, the anticancer effect of CA was evaluated both in vitro and in vivo. Results: Cancer cells treated with CA showed reduced proliferation rate, migration/invasion ability, and mitochondrial ATP production. Treating cancer cells with CA resulted in elevated SUMO1 expression through acting on its 3'UTR and stabilizing the mRNA. The increased SUMO1 caused c-Myc sumoylation, miR-17 family downregulation, and p21 upregulation leading to G0/G1 arrest and maturation phenotype. CA altered the expression of differentiation-related genes in cancer cells but not in normal cells. It inhibited hepatoma and lung cancer growth in tumor-bearing mice and prevented new tumor development in naïve mice. In glioma cells, CA increased expression of specific differentiation biomarkers Tuj1 and GFAP inducing differentiation and reducing sphere formation. The therapeutic efficacy of CA in glioma cells was comparable to that of temozolomide. CA was detectable both in the blood and brain when administered intraperitoneally in animals. Most importantly, CA was safe even at very high doses. CONCLUSION: CA might be a safe and effective differentiation-inducer for cancer therapy. "Educating" cancer cells to differentiate, rather than killing them, could be a novel therapeutic strategy for cancer.


Assuntos
Antineoplásicos/farmacologia , Carcinoma Hepatocelular/tratamento farmacológico , Diferenciação Celular/efeitos dos fármacos , Ácido Clorogênico/farmacologia , Glioma/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Células A549 , Trifosfato de Adenosina/metabolismo , Animais , Antineoplásicos/uso terapêutico , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Transformação Celular Neoplásica/efeitos dos fármacos , Ácido Clorogênico/uso terapêutico , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Pontos de Checagem da Fase G1 do Ciclo Celular , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos NOD , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Ratos , Ratos Wistar , Proteína SUMO-1/metabolismo
9.
Am J Hypertens ; 32(11): 1109-1117, 2019 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-31350549

RESUMO

BACKGROUND: Pulmonary arterial hypertension (PAH) is a severe progressive disease with systemic metabolic dysregulation. Monocrotaline (MCT)-induced and hypoxia-induced pulmonary hypertension (PH) rodent models are the most widely used preclinical models, however, whether or not these preclinical models recapitulate metabolomic profiles of PAH patients remain unclear. METHODS: In this study, a targeted metabolomics panel of 126 small molecule metabolites was conducted. We applied it to the plasma of the 2 preclinical rodent models of PH and 30 idiopathic pulmonary arterial hypertension (IPAH) patients as well as 30 healthy controls to comparatively assess the metabolomic profiles of PAH patients and rodent models. RESULTS: Significantly different metabolomics profiling and pathways were shown among the 2 classical rodent models and IPAH patients. Pathway analysis demonstrated that methionine metabolism and urea cycle metabolism were the most significant pathway involved in the pathogenesis of hypoxia-induced PH model and MCT-induced model, respectively, and both of them were also observed in the dysregulated pathways in IPAH patients. CONCLUSIONS: These 2 models may develop PAH through different metabolomic pathways and each of the 2 classical PH model resembles IPAH patients in certain aspects.


Assuntos
Hipertensão Pulmonar Primária Familiar/sangue , Hipertensão Pulmonar/sangue , Metabolômica , Metionina/sangue , Ureia/sangue , Adulto , Animais , Biomarcadores/sangue , Estudos de Casos e Controles , Modelos Animais de Doenças , Hipertensão Pulmonar Primária Familiar/diagnóstico , Hipertensão Pulmonar Primária Familiar/etiologia , Feminino , Humanos , Hipertensão Pulmonar/diagnóstico , Hipertensão Pulmonar/etiologia , Hipóxia/complicações , Masculino , Monocrotalina , Ratos Sprague-Dawley
10.
Anal Chem ; 91(7): 4504-4512, 2019 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-30840439

RESUMO

Cholesteryl esters (CEs) are formed by the 3-hydroxyl group of cholesterol and a fatty acyl chain through an ester bond and function as a biologically inert storage form of cholesterol. Abnormal CE levels are often related to various diseases, particularly hyperlipidemia and atherosclerosis. Herein, we developed a mathematical model-assisted ultrahigh performance liquid chromatography-mass spectrometry (UHPLC-MS) method for the untargeted identification to targeted quantification of CEs in plasma, different density lipoprotein samples from humans, rats, and golden hamsters. Using UHPLC-quadrupole-time-of-flight mass spectrometry (UHPLC-QTOF-MS), 81 CE candidates were detected in the above samples, of which 24 CEs were reported in the Human Metabolome Database and 57 CEs were newly identified based on an in-house database of theoretically possible CEs, including the computationally generated precursor ion m/ z mass of CE, carbon number and double bond numbers of the fatty acyl chain. Then three mathematical models based on the characteristic chromatographic retention behavior related to structural features were established and validated using commercial and synthetic CE standards. The mathematical model-assisted UHPLC-MS/MS strategy was proposed to provide a global profiling and identification of CEs, especially unknown CEs. With the efficient strategy, 74 CEs in the plasma of golden hamsters were identified and then quantified in normal and hyperlipidemic golden hamsters by dynamic multiple reaction monitoring (dMRM). A total of 21 CEs among 35 shared potential biomarkers were newly found for hyperlipidemia. Our work will contribute to the in-depth study of the functions of CEs and the discovery of disease biomarkers.


Assuntos
Ésteres do Colesterol/análise , Hiperlipidemias/metabolismo , Modelos Teóricos , Espectrometria de Massas em Tandem/métodos , Animais , Biomarcadores/análise , Biomarcadores/sangue , Ésteres do Colesterol/sangue , Cromatografia Líquida de Alta Pressão , Cricetinae , Dieta Hiperlipídica , Modelos Animais de Doenças , Hiperlipidemias/patologia , Limite de Detecção , Mesocricetus , Análise de Componente Principal
11.
Dermatitis ; 29(4): 219-222, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29939855

RESUMO

BACKGROUND: Atopic dermatitis (AD) is a common disease, which involves a disruption of the skin barrier function. Skin ceramide (CER) composition, which plays crucial roles in maintaining the barrier function of the stratum corneum, is changed in patients with AD. OBJECTIVE: The aim of this study was to identify and quantify skin CER subclasses in association with disease severity in pediatric patients with AD. METHODS: Two hundred thirteen patients were entered into the observational study. We compared their CER profiles using normal-phase high-performance liquid chromatography coupled with dynamic multiple reaction monitoring mass spectrometry. RESULTS: In total, 12 subclasses of CERs were identified. We found that 2 subclasses, that is, CER[AS] and CER[NS], were elevated (P = 0.007 and 0.012, respectively) and correlated with Severity Scoring of Atopic Dermatitis (P = 0.004 and 0.004, respectively). CONCLUSIONS: Skin CER abundances are changed in children with AD compared with control subjects.


Assuntos
Ceramidas/análise , Dermatite Atópica/fisiopatologia , Pele/química , Adolescente , Adulto , Idoso , Ceramidas/classificação , Criança , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Adulto Jovem
12.
Syst Biol Reprod Med ; 64(4): 266-273, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29577757

RESUMO

The objective of this study was to explore the association of sperm mitochondrial ND2 (MT-ND2) gene variants with total fertilization failure (TFF). A retrospective comparative study of 246 cases of fresh in vitro fertilization (IVF) cycles or half-intracytoplasmic sperm injection cycles in the Han Chinese population was performed from July 2011 to May 2017. A total of 59 cases undergoing TFF, and 187 control cases with normal fertilization (fertilization rates >50%) were included. The sperm mitochondrial genovariation was determined using nested sequencing. A total of 32 homoplasmic variants and 47 heteroplasmic variants of MT-ND2 gene were observed in this study. There were no significant differences in the frequencies of the 32 homoplasmic variants of MT-ND2 gene between the TFF and control groups. A total of 53 pair-wise comparisons were performed, and the general characteristics of the IVF failure and control subjects were adjusted in logistic models. Data suggested that there were no significant differences in the frequencies of point 4914, 5320, and 5426 heteroplasmic variants of MT-ND2 gene between the TFF and control groups. In addition, no significant difference was observed in the frequency of mtDNA haplogroup D or haplogroup G between the IVF failure group and the normal fertilization group. This study suggests that the MT-ND2 gene variants might not be associated with TFF. ABBREVIATIONS: ATP: adenosine triphosphate; dNTP: deoxy-ribonucleoside triphosphate; FADH2: flavin adenine dinucleotide; FDR: false discovery rate; FSH: follicle-stimulating hormone; IVF: in vitro fertilization; LH: luteinizing hormone; MTATP6: mitochondrially encoded ATP synthase 6; MTCYB: mitochondrially encoded cytochrome b; mtDNA: mitochondrial DNA; MT-ND2: mitochondrial ND2; NADH: nicotinamide adenine dinucleotide; ND2: NADH dehydrogenase subunit 2; OXPHOS: oxidative phosphorylation; PCR: single nucleotide polymorphisms; SNPs: single nucleotide polymorphisms; TFF: total fertilization failure.


Assuntos
DNA Mitocondrial/genética , Fertilização In Vitro , Infertilidade Masculina/genética , NADH Desidrogenase/genética , Adulto , Feminino , Humanos , Masculino , Estudos Retrospectivos
13.
Chin J Integr Med ; 24(2): 117-124, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28000096

RESUMO

OBJECTIVE: To investigate the relationship between tissue distributions of modified Wuzi Yanzong prescription (, MWP) in rats and meridian tropism theory. METHODS: A high-performance liquid chromatography with Fourier transform-mass spectrometry (HPLC-FT) method was used to identify the metabolites of MWP in different tissues of rats after continued oral administration of MWP for 7 days. The relationship between MWP and meridian tropism theory was studied according to the tissue distributions of the metabolites of MWP in rats and the relevant literature. RESULTS: Nineteen metabolites, mainly flavanoid compounds, were detected in the different rat tissues and classified to each herb in MWP. Further, it was able to establish that the tissue distributions of the metabolites of MWP were consistent with the descriptions of meridian tropism of MWP available in literature, this result might be useful in clarifying the mechanism of MWP on meridian tropism. In the long run, these data might provide scientific evidence of the meridian tropism theory to further promote the reasonable, effective utilization, and modernization of Chinese medicine. CONCLUSION: The tissue distributions of MWP in vivo were consistent with the descriptions of meridian tropism of MWP.


Assuntos
Prescrições de Medicamentos , Medicamentos de Ervas Chinesas/farmacologia , Meridianos , Modelos Biológicos , Animais , Medicamentos de Ervas Chinesas/administração & dosagem , Masculino , Metaboloma , Ratos Wistar , Distribuição Tecidual/efeitos dos fármacos
14.
J Sep Sci ; 41(1): 351-372, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28859259

RESUMO

Lipids, which have a core function in energy storage, signalling and biofilm structures, play important roles in a variety of cellular processes because of the great diversity of their structural and physiochemical properties. Lipidomics is the large-scale profiling and quantification of biogenic lipid molecules, the comprehensive study of their pathways and the interpretation of their physiological significance based on analytical chemistry and statistical analysis. Lipidomics will not only provide insight into the physiological functions of lipid molecules but will also provide an approach to discovering important biomarkers for diagnosis or treatment of human diseases. Mass-spectrometry-based analytical techniques are currently the most widely used and most effective tools for lipid profiling and quantification. In this review, the field of mass-spectrometry-based lipidomics was discussed. Recent progress in all essential steps in lipidomics was carefully discussed in this review, including lipid extraction strategies, separation techniques and mass-spectrometry-based analytical and quantitative methods in lipidomics. We also focused on novel resolution strategies for difficult problems in determining C=C bond positions in lipidomics. Finally, new technologies that were developed in recent years including single-cell lipidomics, flux-based lipidomics and multiomics technologies were also reviewed.


Assuntos
Lipídeos/isolamento & purificação , Espectrometria de Massas , Metabolômica , Animais , Biomarcadores/análise , Cromatografia , Humanos , Cinética , Metabolismo dos Lipídeos , Lipídeos/química , Ozônio
15.
Anal Chem ; 89(14): 7808-7816, 2017 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-28643517

RESUMO

Acidic glycosphingolipids (AGSLs), which mainly consist of ganglioside and sulfatide moieties, are highly concentrated in the central nervous system. Comprehensive profiling of AGSLs has historically been challenging because of their high complexity and the lack of standards. In this study, a novel strategy was developed to comprehensively profile AGSLs using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Ganglioside isomers with different glycan chains such as GD1a/GD1b were completely separated on a C18 column for the first time to our knowledge, facilitated by the addition of formic acid in the mobile phase. A mathematical model was established to predict the retention times (RTs) of all theoretically possible AGSLs on the basis of the good logarithmic relationship between the ceramide carbon numbers of the AGSLs in the reference material and their RTs. A data set was created of 571 theoretically possible AGSLs, including the ceramide carbon numbers, RTs, and high-resolution quasi-molecular ions. A novel fast identification strategy was established for global AGSL profiling by comparing the high-resolution quasi-molecular ions and RTs of the tested peaks to those in the data set of 571 AGSLs. Using this strategy, 199 AGSL candidates were identified in rat brain tissue. MS/MS fragments were further collected for these 199 candidates to confirm their identity as AGSLs. This novel strategy was employed to profile AGSLs in brain tissue samples from control rats and model rats with bilateral common carotid artery (2-VO) cerebral ischemia. Forty AGSLs were significantly different between the control and model groups, and these differences were further interpreted.

16.
Biochem Biophys Res Commun ; 488(1): 109-115, 2017 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-28479244

RESUMO

To investigate the effects of the PI3K inhibitors on the differentiation of insulin-producing cells derived from human embryonic stem cells. Here, we report that human embryonic stem cells induced by phosphatidylinositol-3-kinase (PI3K) p110ß inhibitors could produce more mature islet-like cells. Findings were validated by immunofluorescence analysis, quantitative real-time PCR, insulin secretion in vitro and cell transplantation for the diabetic SCID mice. Immunofluorescence analysis revealed that unihormonal insulin-positive cells were predominant in cultures with rare polyhormonal cells. Real-time PCR data showed that islet-like cells expressed key markers of pancreatic endocrine hormones and mature pancreatic ß cells including MAFA. Furthermore, this study showed that the expression of most pancreatic endocrine hormones was similar between groups treated with the LY294002 (nonselective PI3K inhibitor) and TGX-221 (PI3K isoform selective inhibitors of class 1ß) derivatives. However, the level of insulin mRNA in TGX-221-treated cells was significantly higher than that in LY294002-treated cells. In addition, islet-like cells displayed glucose-stimulated insulin secretion in vitro. After transplantation, islet-like cells improved glycaemic control and ameliorated the survival outcome in diabetic mice. This study demonstrated an important role for PI3K p110ß in regulating the differentiation and maturation of islet-like cells derived from human embryonic stem cells.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Classe I de Fosfatidilinositol 3-Quinases/antagonistas & inibidores , Células-Tronco Embrionárias Humanas/citologia , Células-Tronco Embrionárias Humanas/efeitos dos fármacos , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/efeitos dos fármacos , Inibidores de Proteínas Quinases/farmacologia , Animais , Células Cultivadas , Cromonas/farmacologia , Classe I de Fosfatidilinositol 3-Quinases/metabolismo , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Ilhotas Pancreáticas/metabolismo , Masculino , Camundongos , Camundongos SCID , Morfolinas/farmacologia , Inibidores de Proteínas Quinases/química , Pirimidinonas/farmacologia , Relação Estrutura-Atividade
17.
Sci Rep ; 7: 40030, 2017 01 09.
Artigo em Inglês | MEDLINE | ID: mdl-28067267

RESUMO

Atmospheric particle is one of the risk factors for respiratory disease; however, their injury mechanisms are poorly understood, and prevention methods are highly desirable. We constructed artificial PM2.5 (aPM2.5) particles according to the size and composition of actual PM2.5 collected in Beijing. Using these artificial particles, we created an inhalation-injury animal model. These aPM2.5 particles simulate the physical and chemical characteristics of the actual PM2.5, and inhalation of the aPM2.5 in rat results in a time-dependent change in lung suggesting a declined lung function, injury from oxidative stress and inflammation in lung. Thus, this aPM2.5-caused injury animal model may mimic that of the pulmonary injury in human exposed to airborne particles. In addition, polydatin (PD), a resveratrol glucoside that is rich in grapes and red wine, was found to significantly decrease the oxidative potential (OP) of aPM2.5 in vitro. Treating the model rats with PD prevented the lung function decline caused by aPM2.5, and reduced the level of oxidative damage in aPM2.5-exposed rats. Moreover, PD inhibited aPM2.5-induced inflammation response, as evidenced by downregulation of white blood cells in bronchoalveolar lavage fluid (BALF), inflammation-related lipids and proinflammation cytokines in lung. These results provide a practical means for self-protection against particulate air pollution.

18.
Anal Chim Acta ; 950: 108-118, 2017 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-27916115

RESUMO

Eicosanoids are signaling molecules mainly oxidized from arachidonic acid (ARA) and eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA). They have attracted increasing attention from the scientists attributing to their essential physiological functions. However, their quantification have long been challenged by the low abundance, high structure similarity, poor stability and limited ionization efficiency. In this paper, an ultra-high performance liquid chromatograph coupled with tandem mass spectrometry (UPLC-MS/MS) strategy was developed for the comprehensive profiling of more than 60 eicosanoids based on an efficient derivatization reagent 2,4-bis(diethylamino)-6-hydrazino-1,3,5-triazine (T3) and general multiple reaction monitoring (MRM) parameters. Carboxylic acid of eicosanoid was converted to amide in 30 min at 4 °C with derivatization yield larger than 99%. Limits of quantitation (LOQs) for derivatized eicosanoids varied from 0.05 to 50 pg depending on their structures. The sensitivities of derivatized eicosanoids were enhanced by 10- to 5000-folds compared to free eicosanoids. Stabilities of T3 modified eicosanoids were also highly improved compared to free eicosanoids. This new method can also be used to quantify eicosanoids in bio-samples using isotopic internal standards with high efficiency and reliability within 19 min. 46 and 50 eicosanoids in rat plasma and heart tissue from control and acute myocardial ischemia (AMI) model rats were respectively profiled and quantitated using this new method. And 24 of 46 and 25 of 50 eicosanoids were found to be significantly changed between control and model groups. The changed eicosanoids related to AMI modeling were further statistically analyzed and interpreted based on eicosanoid metabolism pathway.


Assuntos
Cromatografia Líquida de Alta Pressão , Eicosanoides/análise , Espectrometria de Massas em Tandem , Animais , Ácidos Carboxílicos , Eicosanoides/sangue , Isquemia Miocárdica/sangue , Miocárdio , Ratos , Reprodutibilidade dos Testes
19.
Anal Chem ; 88(15): 7762-8, 2016 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-27397858

RESUMO

Fatty aldehydes are crucial substances that mediate a wide range of vital physiological functions, particularly lipid peroxidation. Fatty aldehydes such as acrolein and 4-hydroxynonenal (4-HNE) are considered potential biomarkers of myocardial ischemia and dementia, but analytical techniques for fatty aldehydes are lacking. In the present study, a comprehensive characterization strategy with high sensitivity and facility for fatty aldehydes based on derivatization and high-performance liquid chromatography-multiple reaction monitoring (HPLC-MRM) was developed. The fatty aldehydes of a biosample were derivatized using 2,4-bis(diethylamino)-6-hydrazino-1,3,5-triazine under mild and efficient reaction conditions at 37 °C for 15 min. The limit of detection (LOD) of the fatty aldehydes varied from 0.1 to 1 pg/mL, depending on the structures of these molecules. General MRM parameters were forged for the analysis of endogenous fatty aldehydes. "Heavy" derivatization reagents with 20 deuterium atoms were synthesized for both the discovery and comprehensive characterization of fatty aldehydes. More than 80 fatty aldehydes were detected in the biosamples. The new strategy was successfully implemented in global fatty aldehyde profiling of plasma and brain tissue of the bilateral common carotid artery (2VO) dementia rat model. Dozens of fatty aldehydes were significantly changed between the control and model groups. These findings further highlight the importance of endogenous fatty aldehydes.


Assuntos
Aldeídos/análise , Cromatografia Líquida de Alta Pressão , Ácidos Graxos/análise , Acroleína/análise , Acroleína/química , Aldeídos/sangue , Aldeídos/química , Animais , Biomarcadores/análise , Biomarcadores/sangue , Encéfalo/metabolismo , Demência/patologia , Deutério/química , Análise Discriminante , Modelos Animais de Doenças , Ácidos Graxos/sangue , Limite de Detecção , Masculino , Análise de Componente Principal , Ratos , Ratos Wistar , Triazinas/química
20.
Anal Bioanal Chem ; 408(24): 6623-36, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27473427

RESUMO

Skin, the largest organ of the human body, serves as the primary barrier to the external environment. Ceramides are one of the main constituents of stratum corneum (SC), playing an important role in skin barrier function. Therefore, comprehensive profiling and quantification of SC ceramide is important. Herein, a new targeted lipidomic method for human SC ceramide profiling and quantification is presented and tested. Normal-phase high-performance liquid chromatography coupled with dynamic multiple reaction monitoring mass spectrometry (NP-HPLC-dMRM-MS) was used to separate ceramides into subclasses and then characterize different ceramides within each subclass on the basis of their characteristics. In total, 483 ceramides were quantified in a single run within 20 min, covering 12 subclasses as well as some glycosylated ceramides not previously reported. Each subclass had typical standard substances (if available) that served to establish representative standard curves and were used for related substances with no standards. Linearity range, limit of quantification (LOQ), limit of detection (LOD), precision, accuracy, stability, and matrix effects were validated. dMRM increased sensitivity and accuracy greatly compared with common MRM (cMRM). This method was successfully applied to the study of human SC from different age groups. A total of 193 potential biomarkers were found to indicate age differences between children and adults. This method is an innovative approach for high-throughput quantification of SC ceramide. Graphical Abstract Method establishment (MRM spectra by the established method) and method application (score scatter plots of authentic samples).


Assuntos
Ceramidas/análise , Cromatografia Líquida de Alta Pressão/métodos , Epiderme/química , Espectrometria de Massas/métodos , Adulto , Criança , Humanos , Limite de Detecção
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...