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1.
Metabolism ; 102: 154002, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31706979

RESUMO

BACKGROUND: AMP-activated protein kinase (AMPK), particularly AMPKα2 isoform, plays a critical role in maintaining cardiac homeostasis. It was reported that sulforaphane (SFN) prevented type 2 diabetes (T2D)-induced cardiomyopathy accompanied by the activation of AMPK; In this study, AMPK's pivotal role in SFN-mediated prevention against T2D-induced cardiomyopathy was tested using global deletion of AMPKα2 gene (AMPKα2-KO) mice. METHODS AND RESULTS: T2D was established by feeding 3-month high-fat diet (HFD) to induce insulin resistance, followed by an intraperitoneal injection of streptozotocin (STZ) to induce mild hyperglycemia in both AMPKα2-KO and wild-type (WT) mice. Then both T2D and control mice were subsequently treated with or without SFN for 3 months while continually feeding HFD or normal diet. Upon completion of the 3-month treatment, five mice from each group were sacrificed as a 3-month time-point (3 M). The rest continued normal diet or HFD until terminating study at the sixth month (6 M) of diabetes. Cardiac function was examined with echocardiography before sacrifice at both 3 M and 6 M. SFN prevented T2D-induced progression of cardiac dysfunction, remodeling (hypertrophy and fibrosis), inflammation, and oxidative damage in wild-type diabetic mice, but not in AMPKα2-KO mice. Mechanistically, SFN prevented T2D-induced cardiomyopathy not only by improving AMPK-mediated lipid metabolic pathways, but also enhancing NRF2 activation via AMPK/AKT/GSK3ß pathway. However, these improving effects of SFN were abolished in AMPKα2-KO diabetic mice. CONCLUSIONS: AMPK is indispensable for the SFN-induced prevention of cardiomyopathy in T2D, and the activation of NRF2 by SFN is mediated by AMPK/AKT/GSK3ß signaling pathways.


Assuntos
Cardiotônicos/farmacologia , Diabetes Mellitus Tipo 2/complicações , Cardiomiopatias Diabéticas/prevenção & controle , Isotiocianatos/farmacologia , Lipidoses/prevenção & controle , Fator 2 Relacionado a NF-E2/fisiologia , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/fisiologia , Animais , Cardiotônicos/uso terapêutico , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Tipo 2/induzido quimicamente , Diabetes Mellitus Tipo 2/tratamento farmacológico , Cardiomiopatias Diabéticas/genética , Cardiomiopatias Diabéticas/patologia , Isotiocianatos/uso terapêutico , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Lipidoses/genética , Lipidoses/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 2 Relacionado a NF-E2/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Estreptozocina
2.
Int J Med Sci ; 16(12): 1593-1603, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31839747

RESUMO

Background: Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease, and its pathogenesis and mechanism are intricate. In the present study, we aimed to evaluate the role of PPAR δ in LPS associated NAFLD and to investigate the signal transduction pathways underlying PPAR δ treatment in vitro. Material and Methods: L02 cells were exposed to palmitic acid (PA) and/or LPS in the absence or presence of PPAR δ inhibition and/or activation. Results: LPS treatment markedly increased lipid deposition, FFA contents, IL-6 and TNF-α levels, and cell apoptosis in PA treatment (NAFLD model). PPAR δ inhibition protects L02 cells against LPS-induced lipidosis and injury. Conversely, the result of PPAR δ activation showed the reverse trend. LPS+PA treatment group significantly decreases the relative expression level of IRS-1, PI3K, AKT, phosphorylation of AKT, TLR-4, MyD88, phosphorylation of IKKα, NF-κB, Bcl-2 and increases the relative expression level of Bax, cleaved caspase 3 and cleaved caspase 8, compared with the cells treated with NAFLD model. PPAR δ inhibition upregulated the related proteins' expression level in insulin resistance and inflammation pathway and downregulated apoptotic relevant proteins. Instead, PPAR δ agonist showed the reverse trend. Conclusion: Our data show that PPAR δ inhibition reduces steatosis, inflammation and apoptosis in LPS-related NAFLD damage, in vitro. PPAR δ may be a potential therapeutic implication for NAFLD.


Assuntos
Fígado Gorduroso/tratamento farmacológico , Lipidoses/tratamento farmacológico , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , PPAR delta/genética , Substâncias Protetoras/farmacologia , Apoptose/efeitos dos fármacos , Células Cultivadas , Fígado Gorduroso/genética , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Hepatócitos/efeitos dos fármacos , Humanos , Lipidoses/genética , Lipidoses/metabolismo , Lipidoses/patologia , Lipídeos/genética , Lipopolissacarídeos/toxicidade , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Hepatopatia Gordurosa não Alcoólica/genética , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , PPAR delta/agonistas , PPAR delta/antagonistas & inibidores , Ácido Palmítico/toxicidade , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/farmacologia , Transdução de Sinais/efeitos dos fármacos , Tiazóis/farmacologia
3.
Anal Bioanal Chem ; 411(30): 8023-8032, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31776643

RESUMO

Within drug development and pre-clinical trials, a common, significant and poorly understood event is the development of drug-induced lipidosis in tissues and cells. In this manuscript, we describe a mass spectrometry imaging strategy, involving repeated analysis of tissue sections by DESI MS, in positive and negative polarities, using MS and MS/MS modes. We present results of the detected distributions of the administered drug, drug metabolites, lipid molecules and a putative marker of lipidosis, di-docosahexaenoyl (22:6)-bis(monoacylglycerol) phosphate (di-22:6-BMP). A range of strategies have previously been reported for detection, isolation and identification of this compound, which is an isomer of di-docosahexaenoic (22:6 n-3) phosphatidylglycerol (di-22:6 PG), a commonly found lipid that acts as a surfactant in lung tissues. We show that MS imaging using MS/MS can be used to differentiate these compounds of identical mass, based upon the different distributions of abundant fragment ions. Registration of images of these fragments, and detected drugs and metabolites, is presented as a new method for studying drug-induced lipidosis in tissues. Graphical abstract.


Assuntos
Biomarcadores/metabolismo , Lipidoses/induzido quimicamente , Pulmão/diagnóstico por imagem , Espectrometria de Massas/métodos , Amiodarona/efeitos adversos , Animais , Antiarrítmicos/efeitos adversos , Masculino , Ratos Wistar , Roedores
4.
Poult Sci ; 98(9): 3950-3962, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-30941423

RESUMO

Hepatic lipidosis (HL) is a well-known disease in fattening and in parent turkey flocks. Among others, dietary effects like (a lack of) essential amino acids (AA) as lipotrophic factors (e.g., methionine) have been considered as potentially predispositing for HL. Several studies have reported abnormal AA profiles in hepatic diseases of humans and other livestock. The ratio of branched-chain amino acids (BCAA) to aromatic amino acids (AAA) in plasma is used to predict hepatic cirrhosis. In this study, the state of supply of AA was investigated by comparing non-affected (NA) animals and those affected by HL. The AA pattern in the liver and blood can provide potential indications of pathogenesis of HL. In cooperation with German poultry veterinarians, 3 cases of HL on 3 different fattening turkey farms were visited (13/14 wk old, "B.U.T. Big 6" and "TP7"). Overall, 73 birds were examined, of which 42 birds suffered from HL and 31 were not affected. Feeding samples of the respective actual feed were taken and analyzed. The selection of animals was carried out (NA randomly) by clinical signs such as apathy and dyspnea and the diagnosis was made at necropsy, which could be confirmed by crude fat content in liver tissue (HL: 309, NA: 155). In liver tissue, the CP and AA contents were lower among animals with HL than among NA (P < 0.05). In blood samples, the sum of AA, ammonia, and urea was more than 3 times higher among animals with HL (431 mg/dL serum) than among NA (114 mg/dL serum; P < 0.01). The ratio of BCAA to AAA was also significantly different between the groups (HL: 0.85, NA: 1.42; P < 0.05). In the case of HL, entire herds were not affected and the "non-affected" ones were comparable with healthy slaughtered animals. There seems to be a clear change in protein and AA metabolism of HL animals, which could lead to an optimization in feeding practice in repeated cases of HL.


Assuntos
Aminoácidos/metabolismo , Lipidoses/veterinária , Hepatopatias/veterinária , Doenças das Aves Domésticas/metabolismo , Perus , Aminoácidos/sangue , Animais , Feminino , Lipidoses/sangue , Lipidoses/etiologia , Lipidoses/metabolismo , Hepatopatias/sangue , Hepatopatias/etiologia , Hepatopatias/metabolismo , Doenças das Aves Domésticas/sangue , Doenças das Aves Domésticas/etiologia
5.
Invest Ophthalmol Vis Sci ; 60(5): 1789-1798, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-31022733

RESUMO

Purpose: To determine major differences in lipid profile between human control and glaucomatous optic nerve. To assess major enzymes in lipid pathway if aberration is revealed for a lipid class by profiling. Methods: Optic nerve (ON) samples were obtained from human cadaveric donors [control (n = 11) and primary open-angle glaucoma (POAG; n = 12)]; the lipids were extracted using Bligh and Dyer methods. Control and glaucoma donors were all Caucasians age 72.3 ± 5.9 and 70.3 ± 10.5 (inclusive of both sexes), respectively. Lipids were extracted after weighing the tissue; the protein amounts in the corresponding aqueous phase of organic solvent extraction were recorded. High-resolution mass spectrometry was performed using a Q-exactive mass spectrometer coupled with an EASY-nLC 1000 liquid chromatograph instrument. Bioinformatics and statistical analysis were performed using LipidSearch v.4.1 and MetaboAnalyst 4.0/STATA 14.2. Protein amounts were determined using Bradford's method. Western blot, ELISA, and immunohistochemistry utilized established protocols and were performed for protein quantification and localization, respectively. Additional donor tissues were utilized for Western blot, ELISA, and immunohistochemistry. Results: Principal component analysis (PCA) placed control and glaucomatous ONs in two distinct groups based on analysis of lipid profiles. Total lipid, total phospholipids, total ceramide, and total sphingolipids were similar (without significant difference) between control and glaucoma. However, we found a significant increase in glucosylsphingosine in glaucoma compared to control samples. We found similar levels of glucocerebrosidase (GBA), ceramide glucosyltransferase (UGCG), decreased nonlysosomal glucocerebrosidase (GBA2), and increased lysosomal and nonlysosomal acylsphingosine amidohydrolase (ASAH1 and ASAH2) levels in glaucomatous ON compared to control. Conclusions: We found significant differences in glucosylsphingosine lipids, consistent with decreased GBA and GBA2 and increased ASAH1 and ASAH2 immunoreactivity in glaucoma, suggesting the potential impairment of sphingolipid enzymatic pathways in lysosomal and nonlysosomal cellular compartments.


Assuntos
Glaucoma de Ângulo Aberto/metabolismo , Metabolismo dos Lipídeos , Lipidoses/metabolismo , Nervo Óptico/metabolismo , Psicosina/análogos & derivados , Ceramidase Ácida/metabolismo , Idoso , Western Blotting , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Feminino , Glucosilceramidase/metabolismo , Glucosiltransferases/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Espectrometria de Massas , Ceramidase Neutra/metabolismo , Psicosina/metabolismo , beta-Glucosidase/metabolismo
6.
Vet Pathol ; 56(2): 282-288, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30244665

RESUMO

The Quaker parrot has been used as a psittacine model to study clinical lipidology and lipid-related disorders. However, while Quaker parrots appear to be anecdotally susceptible to a variety of spontaneous dyslipidemic disorders and lesions caused by excess lipid accumulation, epidemiologic data are lacking. A multicenter retrospective study on 652 pathology submissions (411 necropsies and 243 biopsies) from Quaker parrots was performed by recording the final pathological diagnoses, age, and sex for each bird. The prevalence of lesions associated with lipid metabolism, such as hepatic lipidosis, atherosclerosis, xanthomas, adipose tumors, coelomic steatitis/steatonecrosis, endogenous lipid pneumonia, and acute pancreatic necrosis/pancreatitis, was reported. Multiple logistic regression models were used to characterize the effects of sex and age on these lesions, and the prevalence of hepatic lipidosis and atherosclerosis was compared to those in a random sample of control psittacine birds. The raw prevalence of atherosclerosis and hepatic lipidosis was 5.6% (95% confidence interval [CI], 3.4%-7.8%) and 21.2% (95% CI, 17.2%-25.1%), respectively. While the prevalence of atherosclerosis was similar to other psittacine species, hepatic lipidosis was more common in Quaker parrots. Quaker parrots also showed a unique susceptibility to acute pancreatic necrosis with a prevalence of 12.9% (95% CI, 9.7%-16.1%). Male parrots were found to be more susceptible than females to lipid accumulation lesions ( P = .0024), including atherosclerosis ( P = .018) and hepatic lipidosis ( P < .001). This retrospective study confirms the high susceptibility of Quaker parrots to lipid-related disorders and presents epidemiological data that may be useful to avian clinicians, pathologists, and researchers using Quaker parrots.


Assuntos
Doenças das Aves/patologia , Transtornos do Metabolismo dos Lipídeos/veterinária , Papagaios , Animais , Aterosclerose/diagnóstico , Aterosclerose/patologia , Aterosclerose/veterinária , Doenças das Aves/diagnóstico , Feminino , Metabolismo dos Lipídeos , Transtornos do Metabolismo dos Lipídeos/diagnóstico , Transtornos do Metabolismo dos Lipídeos/patologia , Lipidoses/diagnóstico , Lipidoses/patologia , Lipidoses/veterinária , Hepatopatias/diagnóstico , Hepatopatias/patologia , Hepatopatias/veterinária , Masculino , Pneumonia Lipoide/diagnóstico , Pneumonia Lipoide/patologia , Pneumonia Lipoide/veterinária , Estudos Retrospectivos , Fatores Sexuais
7.
Pharmacol Res ; 141: 189-200, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30593851

RESUMO

Topiramate is an anticonvulsant drug also prescribed for migraine prophylaxis that acts through several mechanisms of action. Several studies indicate that topiramate induces weight loss and a moderate reduction of plasma lipids and glucose. Based on these favourable metabolic effects, aim of this study was to evaluate if topiramate could modulate atherosclerosis development and protect target organs of dysmetabolic conditions. Thirty apoE-deficient mice were divided into three groups and fed for 12 weeks a high fat diet (Control) or the same diet containing topiramate at 0.125% and 0.250%. Body weight, water and food intake were monitored throughout the study. Plasma lipids and glucose levels were measured and a glucose tolerance test was performed. Atherosclerosis development was evaluated in the whole aorta and at the aortic sinus. Histological analysis of liver, kidney and adipose tissue was performed. Topiramate did not affect weight gain and food intake. Glucose tolerance and plasma lipids were not changed and, in turn, atherosclerosis development was not different among groups. Topiramate did not modify liver and adipose tissue histology. Conversely, in the kidneys, the treatment reduced the occurrence of glomerular lipidosis by decreasing foam cells accumulation and reducing the expression of inflammatory markers. Blood urea nitrogen levels were also reduced by treatment. Our results indicate that topiramate does not affect atherosclerosis development, but preserves kidney structure and function. The study suggests that topiramate could be investigated in drug repurposing studies for the treatment of glomerular lipidosis.


Assuntos
Rim/efeitos dos fármacos , Lipidoses/prevenção & controle , Substâncias Protetoras/farmacologia , Topiramato/farmacologia , Animais , Aterosclerose/metabolismo , Aterosclerose/patologia , Glicemia/análise , Dieta Hiperlipídica , Feminino , Rim/metabolismo , Rim/patologia , Lipidoses/metabolismo , Lipidoses/patologia , Lipídeos/sangue , Camundongos Knockout para ApoE
8.
Sci Total Environ ; 654: 284-291, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30445328

RESUMO

Metals and metalloids are priority contaminants due to their non-degradable and bioaccumulative nature, and their ability to regulate and perturb diverse physiological processes in various species. Metal(loid)s are known to cause oxidative stress through production of reactive oxygen species (ROS), thus related endpoints like lipid peroxidation (LPO) have received considerable attention as biomarkers of exposure. However, the implications of metal(loid) toxicity including LPO on actual lipid profiles of species inhabiting contaminated systems are poorly understood. Here we applied Nuclear Magnetic Resonance (NMR) spectroscopy for untargeted lipidomics of mosquitofish (Gambusia holbrooki) collected from reference and metal(loid)-contaminated wetlands. We measured a range of trace elements in water and fish using inductively coupled plasma - mass spectrometry (ICP-MS), and interpreted site differences in the lipid profiles of mosquitofish in the context of known physiological responses to sub-lethal metal(loid) exposure. Results indicate deregulation of cellular membrane lipids (i.e., glycerophospholipids, cholesterol and sphingolipids) and increased energy storage molecules (i.e., triacylglycerols and fatty acids) in fish from the contaminated wetland. These responses are consistent with the recognised induction of oxidative stress pathways in organisms exposed to metal(loid)s and could also be symptomatic of mitochondrial dysfunction and endocrine disruption. It is difficult to attribute metal(loid)s as the sole factor causing differences between wetlands, and a more controlled experimental approach is therefore warranted to further explore mechanistic pathways. Nevertheless, our study highlights the benefits of untargeted 1H NMR-based lipidomics as a relatively fast and simple approach for field-scale assessment and monitoring of organisms inhabiting metal(loid) contaminated environments.


Assuntos
Antimônio/análise , Arsênico/análise , Membrana Celular/metabolismo , Ciprinodontiformes/metabolismo , Metabolismo Energético/efeitos dos fármacos , Monitoramento Ambiental/métodos , Metabolismo dos Lipídeos/efeitos dos fármacos , Poluentes Químicos da Água/análise , Áreas Alagadas , Animais , Antimônio/toxicidade , Lipidoses , New South Wales , Ressonância Magnética Nuclear Biomolecular , Poluentes Químicos da Água/toxicidade , Qualidade da Água
9.
Toxicol Pathol ; 47(1): 26-34, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30373479

RESUMO

Cationic amphiphilic drugs (CADs) can induce phospholipidosis (PLD) in organs/tissues. Several ophthalmic pharmaceuticals containing CADs are marketed and used in children. To investigate the effect of PLD on the developing cornea, chloroquine and amiodarone, which are representative CADs, were applied topically to the eyes of juvenile rabbits, and the effects in juvenile rabbits were compared with those in young adult rabbits. Diffuse corneal cloudiness was observed in chloroquine- and amiodarone-treated eyes. Histopathologically, vacuolation was observed in the corneal epithelium and keratocytes. On ultrastructural examination, these vacuoles contained multilamellar inclusion bodies, which are a characteristic of PLD. The size of the vacuoles in the corneal epithelium was reduced in juveniles compared with young adults. Cytoplasmic lamellar bodies and exocytosis in the corneal endothelium were observed in young adult rabbits but not in juvenile rabbits. This study revealed that topical application of chloroquine or amiodarone induces corneal PLD in juvenile and young adult rabbits. Corneal endothelial changes occurred only in young adult rabbits, but ophthalmological changes were similar between juveniles and young adults. The results of the study suggest that the effects of corneal PLD were similar among age groups based on risk assessment.


Assuntos
Envelhecimento/metabolismo , Amiodarona/toxicidade , Cloroquina/toxicidade , Córnea/efeitos dos fármacos , Lipidoses/induzido quimicamente , Fosfolipídeos/metabolismo , Administração Oftálmica , Envelhecimento/patologia , Animais , Córnea/metabolismo , Córnea/ultraestrutura , Modelos Animais de Doenças , Feminino , Corpos de Inclusão/metabolismo , Instilação de Medicamentos , Lipidoses/metabolismo , Lipidoses/patologia , Masculino , Coelhos
10.
Vet Clin North Am Exot Anim Pract ; 22(1): 97-107, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30454765

RESUMO

Vision is essential for amphibians, so a healthy ocular surface is critically important. There are ocular surface abnormalities that occur predominantly in captive animals, such as corneal lipidosis, whereas others, such as UV-induced trauma or infectious and parasitic conditions, may be critical to survival for animals in the wild. It is believed that inherited defects are going to be seen in small captive populations but it may be that confined wild groups of amphibians can be just as severely affected. Anything that blinds an animal severely affects its life changes.


Assuntos
Anfíbios , Doenças da Córnea/veterinária , Lipidoses/veterinária , Animais , Córnea/anatomia & histologia , Córnea/patologia , Córnea/fisiologia , Doenças da Córnea/patologia , Lipidoses/patologia , Visão Ocular
11.
Am J Pathol ; 188(9): 1993-2003, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29981744

RESUMO

Drug-induced phospholipidosis is a lysosomal storage disorder characterized by the excess accumulation of tissue phospholipids. Although azithromycin can be used to induce phospholipidosis, no experimental studies evaluating the relationship between drug accumulation and phospholipid localization have been performed. In this study, azithromycin was orally administered to rats for 7 days, and the relationship between drug and phospholipid accumulation was performed using imaging mass microscopy. The administration of azithromycin induced tubular epithelial vacuolation in the inner stripe of the outer medulla of the kidney, consistent with the lamellar bodies that are typical manifestations of drug-induced phospholipidosis. Azithromycin and phospholipid tissue levels were extensively elevated in the kidneys of azithromycin-treated rats. Imaging mass microscopy revealed that both azithromycin and its metabolites were found in the kidneys of azithromycin-treated rats but not in control animals. The vacuolated areas of the kidneys were primarily found in the inner stripe of the outer medulla, consistent with the areas of high azithromycin concentration. Azithromycin was colocalized with several phospholipids-phosphatidylinositol (18:0/20:4), phosphatidylethanolamine (18:0/20:4 and 16:0/20:4), and possibly didocosahexaenoyl (C22:6)-bis(monoacylglycerol) phosphate, a putative biomarker of drug-induced phospholipidosis. In summary, we found correlations between regions of kidney damage and the accumulation of azithromycin, its metabolites, and phospholipids using imaging mass microscopy. Such analyses may help reveal the mechanism and identify putative biomarkers of drug-induced phospholipidosis.


Assuntos
Azitromicina/toxicidade , Nefropatias/patologia , Lipidoses/patologia , Espectrometria de Massas/métodos , Microscopia Eletrônica de Transmissão/métodos , Fosfolipídeos/metabolismo , Animais , Antibacterianos/toxicidade , Processamento de Imagem Assistida por Computador , Nefropatias/induzido quimicamente , Nefropatias/complicações , Nefropatias/metabolismo , Lipidoses/induzido quimicamente , Lipidoses/complicações , Lipidoses/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley
12.
J Clin Endocrinol Metab ; 103(9): 3394-3404, 2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29889238

RESUMO

Context: Menopause is associated with an increased incidence of insulin resistance and diabetes. Objective: The aim of this study was to explore the lipid deposition in liver and skeletal muscle and investigate the association with insulin sensitivity in postmenopausal and premenopausal women. Design and Setting: Single-center cross-sectional study of 55 healthy women between 45 and 60 years of age. We measured lipid deposition in the liver with magnetic resonance spectroscopy, intramuscular and intra-abdominal lipid deposition with MRI, body composition with a dual-energy X-ray absorptiometry scan, and insulin sensitivity with the composite Matsuda Index. Outcome Measures: We studied the association between fat distribution, ectopic lipid deposition, and insulin sensitivity in pre- and postmenopausal women. Results: Postmenopausal women had an increased lipid deposition in the liver [0.68% (0.44 to 0.99) vs 0.49% (0.38 to 0.64), P = 0.01] and skeletal muscle [3% (2 to 4) vs 2% (1 to 3), P = 0.001] and had a 28% lower Matsuda insulin sensitivity index during an oral glucose tolerance test (6.31 ± 3.48 vs 8.78 ± 4.67, P = 0.05) compared with premenopausal women. Total fat mass and leg fat mass were stronger predictors of ectopic lipid deposition, and visceral fat mass was a stronger predictor of both ectopic lipid deposition and insulin resistance in postmenopausal women compared with premenopausal women. Conclusions: For a given subcutaneous and visceral fat depot size, postmenopausal women show increased ectopic lipid deposition and insulin resistance compared with premenopausal women. It is suggested that lipid deposition in liver and skeletal muscle may represent important mechanistic links between the changes in fat depots and the increased incidence of insulin resistance seen after menopause.


Assuntos
Resistência à Insulina/fisiologia , Metabolismo dos Lipídeos/fisiologia , Lipidoses/metabolismo , Pós-Menopausa/metabolismo , Absorciometria de Fóton , Composição Corporal , Estudos Transversais , Feminino , Teste de Tolerância a Glucose , Humanos , Gordura Intra-Abdominal/patologia , Fígado/metabolismo , Imagem por Ressonância Magnética , Pessoa de Meia-Idade , Músculo Esquelético/metabolismo , Pré-Menopausa
13.
BMC Nephrol ; 19(1): 53, 2018 03 06.
Artigo em Inglês | MEDLINE | ID: mdl-29510679

RESUMO

BACKGROUND: It is well-recognized that injection of iodinated radiographic contrast media (CM) sometimes causes acute renal injury via multiple mechanisms, such as vasoconstriction, toxicity on glomerular endothelium and tubular epithelium and so forth. CASE PRESENTATION: A 51-year-old man developed acute renal injury with proteinuria after CM administration. To our surprise, in his renal biopsy sample the myelin figure like structure was observed in glomerular endothelium and mesangial cells by transmission electron microscopy. However the patient didn't has any clinic clues of Fabry disease and other lysosomal storage disorders. Moreover in vitro cultured glomerular endothelial and mesangial cells we found CM triggers lipid aggregation along with the increased CD36 and decreased ABCA1 abundance. Thus this patient was administrated statin to correct the aberrant lipid trafficking, 2 months later at his next visit we found his renal function partially recovered with reduced proteinuria. CONCLUSIONS: Besides the well-known underlying mechanisms, CM may cause renal impairment by triggering the dysregulated transportation of lipid. Furthermore statin is suggested to be a very promising medicine to decrease side effects of CM.


Assuntos
Lesão Renal Aguda/induzido quimicamente , Meios de Contraste/efeitos adversos , Glomérulos Renais/efeitos dos fármacos , Lipidoses/induzido quimicamente , Células Mesangiais/efeitos dos fármacos , Lesão Renal Aguda/patologia , Humanos , Glomérulos Renais/patologia , Glomérulos Renais/ultraestrutura , Lipidoses/patologia , Masculino , Células Mesangiais/patologia , Células Mesangiais/ultraestrutura , Pessoa de Meia-Idade
14.
Toxicol Lett ; 284: 184-194, 2018 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-29248575

RESUMO

Phospholipidosis is a metabolic disorder characterized by intracellular accumulation of phospholipids. It can be caused by short-term or chronic exposure to cationic amphiphilic drugs (CADs). These compounds bind to phospholipids, leading to inhibition of their degradation and consequently to their accumulation in lysosomes. Drug-induced phospholipidosis (DIPL) is frequently at the basis of discontinuation of drug development and post-market drug withdrawal. Therefore, reliable human-relevant in vitro models must be developed to speed up the identification of compounds that are potential inducers of phospholipidosis. Here, hepatic cells derived from human skin (hSKP-HPC) were evaluated as an in vitro model for DIPL. These cells were exposed over time to amiodarone, a CAD known to induce phospholipidosis in humans. Transmission electron microscopy revealed the formation of the typical lamellar inclusions in the cell cytoplasm. Increase of phospholipids was already detected after 24 h exposure to amiodarone, whereas a significant increase of neutral lipid vesicles could be observed after 72 h. At the transcriptional level, the modulation of genes involved in DIPL was detected. These results provide a valuable indication of the applicability of hSKP-HPC for the quick assessment of drug-induced phospholipidosis in vitro, early in the drug development process.


Assuntos
Avaliação Pré-Clínica de Medicamentos/métodos , Hepatócitos/efeitos dos fármacos , Lipidoses/induzido quimicamente , Fosfolipídeos/metabolismo , Pele/citologia , Células-Tronco/citologia , Amiodarona/toxicidade , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Citometria de Fluxo , Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Hepatócitos/ultraestrutura , Humanos , Lipidoses/genética , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Masculino , Fosfolipídeos/genética
15.
Mol Pharm ; 14(12): 4362-4373, 2017 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-29099189

RESUMO

Drug induced phospholipidosis (PLD) may be observed in the preclinical phase of drug development and pose strategic questions. As lysosomes have a central role in pathogenesis of PLD, assessment of lysosomal concentrations is important for understanding the pharmacokinetic basis of PLD manifestation and forecast of potential clinical appearance. Herein we present a systematic approach to provide insight into tissue-specific PLD by evaluation of unbound intracellular and lysosomal (reflecting acidic organelles) concentrations of two structurally related diprotic amines, GRT1 and GRT2. Their intratissue distribution was assessed using brain and lung slice assays. GRT1 induced PLD both in vitro and in vivo. GRT1 showed a high intracellular accumulation that was more pronounced in the lung, but did not cause cerebral PLD due to its effective efflux at the blood-brain barrier. Compared to GRT1, GRT2 revealed higher interstitial fluid concentrations in lung and brain, but more than 30-fold lower lysosomal trapping capacity. No signs of PLD were seen with GRT2. The different profile of GRT2 relative to GRT1 is due to a structural change resulting in a reduced basicity of one amino group. Hence, by distinct chemical modifications, undesired lysosomal trapping can be separated from desired drug delivery into different organs. In summary, assessment of intracellular unbound concentrations was instrumental in delineating the intercompound and intertissue differences in PLD induction in vivo and could be applied for identification of potential lysosomotropic compounds in drug development.


Assuntos
Diaminas/farmacologia , Lipidoses/induzido quimicamente , Modelos Biológicos , Animais , Encéfalo/metabolismo , Química Farmacêutica , Líquido Extracelular/metabolismo , Feminino , Células Hep G2 , Humanos , Pulmão/metabolismo , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Masculino , Modelos Animais , Modelos Químicos , Fosfolipídeos/metabolismo , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Distribuição Tecidual
16.
Mol Pharm ; 14(12): 4346-4352, 2017 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-29077420

RESUMO

The drug-induced accumulation of phospholipids in lysosomes of various tissues is predominantly observed in regular repeat dose studies, often after prolonged exposure, and further investigated in mechanistic studies prior to candidate nomination. The finding can cause delays in the discovery process inflicting high costs to the affected projects. This article presents an in vitro imaging-based method for early detection of phospholipidosis liability and a hybrid approach for early detection and risk mitigation of phospolipidosis utilizing the in vitro readout with in silico model prediction. A set of reference compounds with phospolipidosis annotation was used as an external validation set yielding accuracies between 77.6% and 85.3% for various in vitro and in silico models, respectively. By means of a small set of chemically diverse known drugs with in vivo phospholipidosis annotation, the advantages of combining different prediction methods to reach an overall improved phospholipidosis prediction will be discussed.


Assuntos
Descoberta de Drogas/métodos , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/prevenção & controle , Lipidoses/induzido quimicamente , Lisossomos/metabolismo , Fosfolipídeos/metabolismo , Animais , Linhagem Celular Tumoral , Biologia Computacional/métodos , Simulação por Computador , Descoberta de Drogas/economia , Avaliação Pré-Clínica de Medicamentos/métodos , Técnicas In Vitro , Aprendizado de Máquina , Microscopia de Fluorescência
17.
J Toxicol Sci ; 42(5): 589-596, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28904294

RESUMO

It is important to consider susceptibility to drug-induced toxicity between animals and humans. Chimeric mice with a humanized liver are expected to predict hepatotoxicity in humans. Drug-induced phospholipidosis (DIPL), in which phospholipids accumulate, is a known entity. In this study, we examined whether chimeric mice can reveal species differences in DIPL. Changes in various phosphatidylcholine (PhC) molecules were investigated in the liver of chimeric mice after administering amiodarone, which induces phospholipidosis. Liquid chromatography-tandem mass spectrometry revealed that levels of PhCs tended to increase in the liver after administration of amiodarone. The liver of chimeric mice consists of human hepatocytes and residual mouse hepatocytes. We used imaging mass spectrometry (IMS) to evaluate the increase of PhCs in human and mouse hepatocytes after administration of amiodarone. IMS visualizes localization of endogenous and exogenous molecules in tissues. The IMS analysis suggested that the localized levels of several PhCs tended to be higher in the human hepatocytes than those in mouse hepatocytes, and PhC levels changed in response to amiodarone. Chimeric mice with a humanized liver will be useful to evaluate species differences in DIPL between mice and humans.


Assuntos
Amiodarona/toxicidade , Hepatócitos/metabolismo , Hepatócitos/transplante , Lipidoses/induzido quimicamente , Lipidoses/metabolismo , Fígado/metabolismo , Fosfatidilcolinas/metabolismo , Fosfolipídeos/metabolismo , Quimeras de Transplante , Animais , Cromatografia Líquida , Humanos , Camundongos , Especificidade da Espécie , Espectrometria de Massas em Tandem
18.
J Toxicol Sci ; 42(5): 641-650, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28904299

RESUMO

The utility of HepaRG cells as an in vitro cell-based assay system for predicting drug-induced phospholipidosis (PLD) was investigated. In experiment 1, 10 PLD-positive compounds and 11 PLD-negative compounds were selected. HepaRG cells were treated with each compound for 48 hr. In experiment 2, loratadine and desloratadine, a major metabolite of loratadine, were used to assess metabolic activation for PLD. HepaRG cells were treated with loratadine and desloratadine in the presence or absence of 500 µM 1-aminobenzotriazole (ABT), a broad CYP inhibitor, for 48 hr. After treatment with compounds in experiments 1 and 2, the relative fluorescence intensity (RFI) was measured using LYSO-ID Red dye to assess the PLD induction. In experiment 1, our cell-based assay system using HepaRG cells exhibited 100% sensitivity and 100% specificity for predicting drug-induced PLD. In experiment 2, loratadine increased the RFI in the PLD assay. However, the increase in the RFI was not observed in co-treatment with loratadine and ABT. In addition, desloratadine increased the RFI in the presence and absence of ABT. These results suggested that metabolic activation of loratadine may contribute to PLD in HepaRG cells. We newly demonstrated that HepaRG cells have a high ability for predicting drug-induced PLD. In addition, we newly showed that HepaRG cells may predict drug-induced PLD mediated by metabolic activation of loratadine. Thus, a cell-based assay system using HepaRG cells is a useful model for predicting drug-induced PLD.


Assuntos
Bioensaio/métodos , Lipidoses/induzido quimicamente , Lipidoses/metabolismo , Fosfolipídeos/metabolismo , Amicacina/toxicidade , Amiodarona/toxicidade , Amitriptilina/toxicidade , Clorpromazina/toxicidade , Feminino , Células Hep G2 , Humanos , Imipramina/efeitos adversos , Loratadina/análogos & derivados , Loratadina/toxicidade , Valor Preditivo dos Testes , Triazóis/toxicidade
19.
Int J Toxicol ; 36(5): 386-394, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28820006

RESUMO

Immunohistochemical staining for the lysosome-associated membrane protein 2 (LAMP-2) has been proposed previously as an alternative to electron microscopy to identify hepatic phospholipidosis. This study used LAMP-2 immunohistochemistry (IHC) to diagnose phospholipidosis in rats exhibiting renal tubular injury. Rats were administered toreforant, a histamine H4 receptor antagonist by oral gavage at a dose of 3, 10, or 100 mg/kg/d for 6 months. Hematoxylin and eosin staining revealed renal tubular epithelial cell vacuolation, hypertrophy, degeneration, and luminal dilation in the 100 mg/kg/d group animals. Renal tubular injury was confirmed using kidney injury marker 1 (KIM-1) IHC. The involvement of phosopholipidosis in the renal injury was investigated by LAMP-2. Adipophilin IHC was included to differentiate phospholipidosis from lipidosis. Increased LAMP-2 staining was observed in the 100 mg/kg/d group animals when compared to vehicle group animals. Lysosome-associated membrane protein-2 staining was most prominent in the outer stripe of the outer medulla where KIM-1 staining was also most prominent. By contrast, adipophilin staining was not increased. Phospholipidosis was also confirmed by electron microscopy. These data support the use of LAMP-2 IHC as a diagnostic tool and suggest an association between phospholipidosis and the renal tubular injury caused by toreforant.


Assuntos
Antagonistas dos Receptores Histamínicos/toxicidade , Nefropatias/diagnóstico , Rim/efeitos dos fármacos , Lipidoses/diagnóstico , Proteína 2 de Membrana Associada ao Lisossomo/metabolismo , Fosfolipídeos/metabolismo , Receptores Histamínicos H4/antagonistas & inibidores , Lesão Renal Aguda , Animais , Moléculas de Adesão Celular/metabolismo , Feminino , Imuno-Histoquímica , Rim/metabolismo , Rim/patologia , Rim/ultraestrutura , Nefropatias/induzido quimicamente , Nefropatias/metabolismo , Nefropatias/patologia , Lipidoses/induzido quimicamente , Lipidoses/metabolismo , Lipidoses/patologia , Proteína 2 de Membrana Associada ao Lisossomo/análise , Masculino , Microscopia Eletrônica de Transmissão , Perilipina-2/análise , Perilipina-2/metabolismo , Ratos Sprague-Dawley
20.
BMC Vet Res ; 13(1): 231, 2017 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-28789691

RESUMO

BACKGROUND: A link between lipid metabolism and disease has been recognized in cats. Since hepatic lipidosis is a frequent disorder in cats, the aim of the current study was to evaluate liver and plasma lipid dimorphism in healthy cats and the effects of gonadectomy on lipid profiling. From six female and six male cats plasma and liver lipid profiles before and after spaying/neutering were assessed and compared to five cats (three neutered male and two spayed female) diagnosed with hepatic lipidosis. RESULTS: Intact female cats had a significantly lower level of plasma triacylglycerides (TAG) and a higher liver level of the long chain polyunsaturated fatty acid arachidonic acid (AA) compared to their neutered state. Both male and female cats with lipidosis had a higher liver, but not plasma TAG level and an increased level of plasma and liver sphingomyelin compared to the healthy cats. CONCLUSION: Although lipid dimorphism in healthy cats resembles that of other species, intact female cats show differences in metabolic configuration that could predispose them to develop hepatic lipidosis. The increased sphingomyelin levels in cats with lipidosis could suggest a potential role in the pathogenesis of hepatic lipidosis in cats.


Assuntos
Doenças do Gato/metabolismo , Metabolismo dos Lipídeos , Lipidoses/veterinária , Fígado/metabolismo , Animais , Ácido Araquidônico/sangue , Doenças do Gato/sangue , Gatos , Feminino , Lipidoses/sangue , Lipidoses/metabolismo , Masculino , Orquiectomia/veterinária , Ovariectomia/veterinária , Fatores Sexuais , Esfingomielinas/sangue , Triglicerídeos/sangue
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