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1.
Int J Mol Med ; 47(4)2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33649807

RESUMO

Excessive lung inflammation caused by endotoxins, including lipopolysaccharide (LPS), mediates the detrimental effects of acute lung injury (ALI), as evidenced by severe alveolar epithelial cell injury. CD40, a member of the tumor necrosis factor receptor superfamily, serves as a central activator in triggering and transducing a series of severe inflammatory events during the pathological processes of ALI. Ginkgolide C (GC) is an efficient and specific inhibitor of CD40. Therefore, the present study aimed to investigate whether GC alleviated LPS­induced ALI, as well as the potential underlying mechanisms. LPS­injured wild­type and CD40 gene conditional knockout mice, and primary cultured alveolar epithelial cells isolated from these mice served as in vivo and in vitro ALI models, respectively. In the present study, histopathological assessment, polymorphonuclear neutrophil (PMN) infiltration, lung injury score, myeloperoxidase activity, wet­to­dry (W/D) weight ratio and hydroxyproline (Hyp) activity were assessed to evaluate lung injury. In addition, immunohistochemistry was performed to evaluate intracellular adhesion molecule­1, vascular cell adhesion molecule­1 and inducible nitric oxide synthase expression levels, and TNF­α, IL­1ß, IL­6 ELISAs and western blotting were conducted to elucidate the signaling pathway. The results demonstrated that GC alleviated LPS­induced lung injury, as evidenced by improvements in ultrastructural characteristics and histopathological alterations of lung tissue, inhibited PMN infiltration, as well as reduced lung injury score, W/D weight ratio and hydroxyproline content. In LPS­injured alveolar epithelial cells, GC significantly reduced IκBα phosphorylation, IKKß activity and NF­κB p65 subunit translocation via downregulating CD40, leading to a significant decrease in downstream inflammatory cytokine levels and protein expression levels. In conclusion, the results of the present study demonstrated that GC displayed a protective effect against LPS­induced ALI via inhibition of the CD40/NF­κB signaling pathway; therefore, the present study suggested that the CD40/NF­κB signaling pathway might serve as a potential therapeutic target for ALI.


Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Antígenos CD40/antagonistas & inibidores , Ginkgolídeos/farmacologia , Quinase I-kappa B/metabolismo , Lactonas/farmacologia , Fator de Transcrição RelA/metabolismo , Lesão Pulmonar Aguda/patologia , Animais , Antígenos CD40/genética , Antígenos CD40/metabolismo , Células Cultivadas , Citocinas/sangue , Hidroxiprolina/metabolismo , Inflamação/tratamento farmacológico , Lipopolissacarídeos/toxicidade , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Peroxidase/metabolismo , Transdução de Sinais/efeitos dos fármacos
2.
Arch Biochem Biophys ; 698: 108727, 2021 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-33333077

RESUMO

Proline utilization A (PutA) proteins are bifunctional proline catabolic enzymes that catalyze the 4-electron oxidation of l-proline to l-glutamate using spatially-separated proline dehydrogenase and l-glutamate-γ-semialdehyde dehydrogenase (GSALDH, a.k.a. ALDH4A1) active sites. The observation that l-proline inhibits both the GSALDH activity of PutA and monofunctional GSALDHs motivated us to study the inhibition of PutA by proline stereoisomers and analogs. Here we report five high-resolution crystal structures of PutA with the following ligands bound in the GSALDH active site: d-proline, trans-4-hydroxy-d-proline, cis-4-hydroxy-d-proline, l-proline, and trans-4-hydroxy-l-proline. Three of the structures are of ternary complexes of the enzyme with an inhibitor and either NAD+ or NADH. To our knowledge, the NADH complex is the first for any GSALDH. The structures reveal a conserved mode of recognition of the inhibitor carboxylate, which results in the pyrrolidine rings of the d- and l-isomers having different orientations and different hydrogen bonding environments. Activity assays show that the compounds are weak inhibitors with millimolar inhibition constants. Curiously, although the inhibitors occupy the aldehyde binding site, kinetic measurements show the inhibition is uncompetitive. Uncompetitive inhibition may involve proline binding to a remote site or to the enzyme-NADH complex. Together, the structural and kinetic data expand our understanding of how proline-like molecules interact with GSALDH, reveal insight into the relationship between stereochemistry and inhibitor affinity, and demonstrate the pitfalls of inferring the mechanism of inhibition from crystal structures alone.


Assuntos
Proteínas de Bactérias/metabolismo , Inibidores Enzimáticos/metabolismo , Glutamato-5-Semialdeído Desidrogenase/metabolismo , Hidroxiprolina/metabolismo , Proteínas de Membrana/metabolismo , Prolina/metabolismo , Proteínas de Bactérias/química , Domínio Catalítico , Cristalografia por Raios X , Inibidores Enzimáticos/química , Glutamato-5-Semialdeído Desidrogenase/química , Hidroxiprolina/química , Proteínas de Membrana/química , Prolina/química , Ligação Proteica , Sinorhizobium meliloti/enzimologia , Estereoisomerismo
3.
Am J Physiol Gastrointest Liver Physiol ; 320(2): G166-G174, 2021 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-33325808

RESUMO

Human carboxylesterase 2 (CES2) has triacylglycerol hydrolase (TGH) activities and plays an important role in lipolysis. In this study, we aim to determine the role of human CES2 in the progression or reversal of steatohepatitis in diet-induced or genetically obese mice. High-fat/high-cholesterol/high-fructose (HFCF) diet-fed C57BL/6 mice or db/db mice were intravenously injected with an adeno-associated virus expressing human CES2 under the control of an albumin promoter. Human CES2 protected against HFCF diet-induced nonalcoholic fatty liver disease (NAFLD) in C57BL/6J mice and reversed steatohepatitis in db/db mice. Human CES2 also improved glucose tolerance and insulin sensitivity. Mechanistically, human CES2 reduced hepatic triglyceride (T) and free fatty acid (FFA) levels by inducing lipolysis and fatty acid oxidation and inhibiting lipogenesis via suppression of sterol regulatory element-binding protein 1. Furthermore, human CES2 overexpression improved mitochondrial respiration and glycolytic function, and inhibited gluconeogenesis, lipid peroxidation, apoptosis, and inflammation. Our data suggest that hepatocyte-specific expression of human CES2 prevents and reverses steatohepatitis. Targeting hepatic CES2 may be an attractive strategy for treatment of NAFLD.NEW & NOTEWORTHY Human CES2 attenuates high-fat/cholesterol/fructose diet-induced steatohepatitis and reverses steatohepatitis in db/db mice. Mechanistically, human CES2 induces lipolysis, fatty acid and glucose oxidation, and inhibits hepatic glucose production, inflammation, lipid oxidation, and apoptosis. Our data suggest that human CES2 may be targeted for treatment of non-alcoholic steatohepatitis (NASH).


Assuntos
Carboxilesterase/metabolismo , Hepatócitos/enzimologia , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Hepatopatia Gordurosa não Alcoólica/terapia , Ácido 3-Hidroxibutírico/sangue , Ácido 3-Hidroxibutírico/metabolismo , Alanina Transaminase/sangue , Alanina Transaminase/metabolismo , Animais , Apoptose/fisiologia , Aspartato Aminotransferases/sangue , Aspartato Aminotransferases/metabolismo , Glicemia , Carboxilesterase/genética , Dieta/efeitos adversos , Hidroxiprolina/sangue , Hidroxiprolina/metabolismo , Metabolismo dos Lipídeos , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Camundongos Transgênicos , Obesidade/induzido quimicamente , Espécies Reativas de Oxigênio/metabolismo
4.
Transl Res ; 224: 16-25, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32504824

RESUMO

Epidemiological studies found that increases in the concentrations of airborne particulate matter (PM) smaller than 10 microns diameter (PM10) in the ambient air due to desert dust outbreaks contribute to global burden of diseases, primarily as a result of increased risk of cardiovascular morbidity and mortality. No studies have investigated the possible association between desert dust inhalation and airway inflammation in patients with ischemic heart disease (IHD). Induced sputum was collected in 38 patients and analyzed to determine markers of airway inflammation (Transforming Growth Factor-ß1 [TGF-ß1] and hydroxyproline) concentrations. For the purpose of the investigation, PM10 and reactive gases concentrations measured in the European Air Quality Network implemented in the Canary Islands were also used. We identified Saharan desert dust using meteorology and dust models. Patients affected by smoking, chronic obstructive pulmonary disease (COPD), asthma, pulmonary abnormalities, acute bronchial or pulmonary disease were excluded. The median of age of patients was 64.71 years (56.35-71.54) and 14 (38.84%) of them were women. TGF-ß1 and hydroxyproline in sputum were highly associated to PM10 inhalation from the Saharan desert. According to a regression model, an increase of 1 µg/m3 of PM10 concentrations due to desert dust, results in an increase of 3.84 pg/gwt of TGF-ß1 (R2 adjusted = 89.69%) and of 0.80 µg/gwt of hydroxyproline (R2 adjusted = 85.28%) in the sputum of patients. The results of this study indicate that the exposure to high PM10 concentrations due to Saharan dust events are associated with intense inflammatory reaction in the airway mucosae of IHD-patients.


Assuntos
Poluição do Ar/análise , Poeira/análise , Exposição Ambiental , Inflamação/etiologia , Pulmão/patologia , Isquemia Miocárdica/complicações , África do Norte , Idoso , Feminino , Humanos , Hidroxiprolina/metabolismo , Masculino , Pessoa de Meia-Idade , Material Particulado/efeitos adversos , Fatores de Risco , Espanha , Escarro , Fator de Crescimento Transformador beta1/metabolismo
5.
Life Sci ; 256: 117893, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32502539

RESUMO

AIMS: To investigate the effect and underlying mechanism of melittin and tripartite motif (TRIM) family in human embryonic lung fibroblast (HELF). MATERIALS AND METHODS: Lentiviral RNA interference vector and lentiviral overexpression vector were constructed and packaged by transfecting 293T cells; the proliferation of HELF was examined using Cell Counting Kit 8; Western blot and qRT-PCR were performed to examine protein and mRNA expression; the interaction with protein phosphatase magnesium-dependent 1A (PPM1A) was examined by Co-immunoprecipitation. KEY FINDINGS: Compared with the control group, the mRNA expression of the TRIM6, TRIM8 and TRIM47 in the IPF group significantly increased. Melittin inhibited the mRNA expression and protein expression levels of TRIM47, the HELF proliferation, the hydroxyproline levels, and the phosphorylation of Smad2/3; the interference of TRIM47 inhibited the protein expression of Vimentin, α-SMA, CTGF, the phosphorylation of Smad2/3 and the synthesis of hydroxyproline; TRIM47 overexpression elevated the phosphorylation of Smad2/3, induced ubiquitination of PPM1A and decreased the expression level of PPM1A, while TRIM47 RNA interference reversed this result. SIGNIFICANCE: Melittin has anti-fibrotic effect in HELF by directly reducing the phosphorylation of Smad2/3 or indirectly reducing the phosphorylation of Smad2/3 by decreasing the expression levels of TRIM47 whose overexpression induces ubiquitination of PPM1A.


Assuntos
Proteínas de Transporte/metabolismo , Fibroblastos/metabolismo , Fibroblastos/patologia , Pulmão/embriologia , Meliteno/farmacologia , Proteínas de Neoplasias/metabolismo , Proteínas Nucleares/metabolismo , Transdução de Sinais , Actinas/metabolismo , Proteínas de Transporte/sangue , Proliferação de Células/efeitos dos fármacos , Fator de Crescimento do Tecido Conjuntivo/genética , Fator de Crescimento do Tecido Conjuntivo/metabolismo , Fibroblastos/efeitos dos fármacos , Fibrose , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Hidroxiprolina/metabolismo , Proteínas de Neoplasias/sangue , Proteínas Nucleares/sangue , Fosforilação/efeitos dos fármacos , Proteína Fosfatase 2C/metabolismo , Interferência de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteínas Smad/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Ubiquitinação/efeitos dos fármacos , Vimentina/metabolismo
6.
Toxicol Lett ; 331: 112-121, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32534005

RESUMO

Roxadustat is the first orally administered, small-molecule hypoxia-inducible factor (HIF) prolyl hydroxylase inhibitor that has been submitted for FDA regulatory approval to treat anemia secondary to chronic kidney diseases. Its usage has also been suggested for pulmonary fibrosis; however, the corresponding therapeutic effects remain to be investigated. The in vitro effects of roxadustat on cobalt chloride (CoCl2)-stimulated pulmonary fibrosis with L929 mouse fibroblasts as well as on an in vivo pulmonary fibrosismice model induced with bleomycin (BLM; intraperitoneal injection, 50 mg/kg twice a week for 4 continuous weeks) were investigated. It found that the proliferation of L929 cells was inhibited and the production of collagen I, collagen III, prolyl hydroxylase domain protein 2 (PHD2), HIF-1α, α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), transforming growth factor-ß1 (TGF-ß1) and p-Smad3 were reduced relative to that in the CoCl2 or BLM group after roxadustat treatment. Roxadustat ameliorated pulmonary fibrosis by reducing the pathology score and collagen deposition as well as decreasing the expression of collagen I, collagen III, PHD2, HIF-1α, α-SMA, CTGF, TGF-ß1 and p-Smad3/Smad3. Our cumulative results demonstrate that roxadustat administration can attenuate experimental pulmonary fibrosis via the inhibition of TGF-ß1/Smad activation.


Assuntos
Fibroblastos/efeitos dos fármacos , Glicina/análogos & derivados , Prolina Dioxigenases do Fator Induzível por Hipóxia/antagonistas & inibidores , Isoquinolinas/uso terapêutico , Pulmão/efeitos dos fármacos , Fibrose Pulmonar/prevenção & controle , Animais , Bleomicina/farmacologia , Técnicas de Cultura de Células , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Fibroblastos/enzimologia , Glicina/farmacologia , Glicina/uso terapêutico , Hidroxiprolina/metabolismo , Isoquinolinas/farmacologia , Pulmão/enzimologia , Pulmão/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fibrose Pulmonar/enzimologia
7.
Life Sci ; 254: 117795, 2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-32417373

RESUMO

AIMS: The primary focus of this study was to explore the effects of cyclic AMP response element-binding protein H (CREBH) on the development of nonalcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: CREBH knockout (KO) and wildtype (WT) mice were averagely divided into a methionine and choline-deficient (MCD) or high fat (HF) diet group and respective chow diet (CD) groups. Mice were sacrificed after 4-week treatment for MCD model and 24-week treatment for HF model. KEY FINDINGS: Characteristics of nonalcoholic steatohepatitis-related liver fibrosis in KO-MCD/HF group were verified by hepatic histological analyses. Compared with WT-MCD/HF group, levels of plasma ALT and hepatic hydroxyproline increased in KO-MCD/HF group. Significantly higher levels of MCP-1, αSMA, Desmin, COL-1, TIMP-1, TGF-ß1, TGF-ß2 were found while MMP-9 and FGF21 mRNA levels decreased in KO-MCD/HF group. There was also a distinct difference of mRNA levels of TNFα, CTGF and CCND1 in KO-HF group compared with controls. Protein levels of MCP-1, BAX, αSMA, COL-1, TGF-ß1 and SMAD2/3 significantly increased in KO-MCD/HF group and CCND1 was also upregulated in KO-HF group compared to their counterparts. SIGNIFICANCE: CREBH knockout may primarily regulate the TGF-ß1 signaling pathway via TGF-ß2 and FGF21 resulting in more severe inflammation and fibrosis in NAFLD.


Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Cirrose Hepática/genética , Hepatopatia Gordurosa não Alcoólica/genética , Fator de Crescimento Transformador beta/metabolismo , Alanina Transaminase/sangue , Animais , Deficiência de Colina , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/deficiência , Dieta Hiperlipídica , Fatores de Crescimento de Fibroblastos/biossíntese , Hidroxiprolina/metabolismo , Lipídeos/sangue , Cirrose Hepática/sangue , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Masculino , Metionina/deficiência , Camundongos , Camundongos Knockout , Hepatopatia Gordurosa não Alcoólica/sangue , Hepatopatia Gordurosa não Alcoólica/metabolismo , Hepatopatia Gordurosa não Alcoólica/patologia , Transdução de Sinais/genética
8.
Int J Nanomedicine ; 15: 2829-2839, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32368057

RESUMO

Objective: To investigate the remineralizing and staining effects of sodium fluoride (NaF) solution with polyethylene glycol-coated silver nanoparticles (PEG-AgNPs) on artificial dentine caries. Materials and Methods: Demineralized human dentine blocks were allocated to three groups. The blocks in group 1 underwent a topical application of a 12% silver diamine fluoride (SDF, 14,150 ppm fluoride) solution. The blocks in group 2 received a topical application of a 2.5% NaF (11,310 ppm fluoride) with PEG-AgNPs (400 ppm silver). The blocks in group 3 received deionized water. All blocks were subjected to pH cycling for 8 days. The surface morphology and cross-sectional features were investigated using scanning electron microscopy (SEM). The color parameters, crystal characteristics, lesion depth, and collagen degradation of the blocks were assessed using digital spectrophotometry, X-ray diffraction (XRD), micro-computed tomography, and spectrophotometry with a hydroxyproline assay, respectively. Results: The SEM showed that dentine collagen was exposed in group 3 but not in groups 1 and 2. The mean lesion depths in groups 1 to 3 were 118±7 µm, 121±14 µm, and 339±20 µm, respectively (groups1,2<3; p<0.001). The data indicated that fluoridated PEG-AgNPs introduced no significant color effect on dentine, but SDF caused distinct discoloration. The XRD indicated that silver chloride was formed in group 1, and fluorapatite was detected in groups 1 and 2. The concentration of hydroxyproline liberated from collagen was significantly less in groups 1 and 2 than in group 3. Conclusion: The use of NaF solution with PEG-AgNPs can remineralize artificial dentine caries and inhibit collagen degradation without causing significant tooth staining.


Assuntos
Cárie Dentária/tratamento farmacológico , Dentina/efeitos dos fármacos , Nanopartículas Metálicas/uso terapêutico , Fluoreto de Sódio/farmacologia , Remineralização Dentária/métodos , Colágeno , Cor , Dentina/metabolismo , Dentina/patologia , Fluoretos/farmacologia , Fluoretos Tópicos/química , Fluoretos Tópicos/farmacologia , Humanos , Hidroxiprolina/metabolismo , Nanopartículas Metálicas/administração & dosagem , Nanopartículas Metálicas/química , Microscopia Eletrônica de Varredura , Dente Serotino/efeitos dos fármacos , Dente Serotino/patologia , Dente Serotino/ultraestrutura , Polietilenoglicóis/química , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/farmacologia , Prata , Compostos de Prata/química , Compostos de Prata/farmacologia , Fluoreto de Sódio/administração & dosagem , Difração de Raios X , Microtomografia por Raio-X
9.
Immunopharmacol Immunotoxicol ; 42(2): 138-146, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32116062

RESUMO

Objectives: Paraquat (PQ) poisoning can induce mitophagy and pulmonary fibrosis. Cyclosporine A (CsA) is an inhibitor of mitophagy. This study aimed at investigating whether CsA could inhibit PQ-induced mitophagy and pulmonary fibrosis in rats.Materials and Methods: Male Sprague-Dawley (SD) rats were treated with vehicle saline (control), 50 mg/kg PQ by gavage alone, or together with different doses of CsA. At 14 days post-induction, the levels of pulmonary fibrosis and PTEN-induced putative kinase 1 (PINK1) and Parkin expression in individual rats and mitochondrial membrane potential (MMP) in lung cells were measured. Moreover, A549 cells were treated with PQ or PQ + CsA for 24 h and the levels of PINK1, Parkin, fibronectin, collagen I and LC3 I and II expression and MMP were examined. Finally, the impact of PINK1 overexpression on the PQ or PQ + CsA-modulated fibronectin and collagen I expression in A549 cells was tested.Results: PQ exposure significantly increased the levels of hydroxyproline and collagen I expression and collagen fiber accumulation in the lung of rats, which were mitigated by CsA treatment. Furthermore, treatment with CsA significantly improved the PQ-decreased MMP and abrogated PQ-upregulated PINK1 and Parkin expression in the lungs of rats. In addition, CsA treatment decreased the PQ-induced fibrosis and mitophagy and PQ-impaired MMP as well as PQ-upregulated PINK1 and Parkin expression in A549 cells. The later effect of CsA was abrogated by PINK1 overexpression in A549 cells.Conclusions: Therefore, CsA can inhibit the PQ-induced mitophagy and pulmonary fibrosis by attenuating the PINK1/Parkin signaling.


Assuntos
Ciclosporina/farmacologia , Pulmão/efeitos dos fármacos , Mitofagia/efeitos dos fármacos , Paraquat/envenenamento , Proteínas Quinases/metabolismo , Fibrose Pulmonar/prevenção & controle , Ubiquitina-Proteína Ligases/metabolismo , Células A549 , Animais , Modelos Animais de Doenças , Humanos , Hidroxiprolina/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Ratos Sprague-Dawley
10.
Amino Acids ; 52(3): 329-360, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32072297

RESUMO

Taurine (a sulfur-containing ß-amino acid), creatine (a metabolite of arginine, glycine and methionine), carnosine (a dipeptide; ß-alanyl-L-histidine), and 4-hydroxyproline (an imino acid; also often referred to as an amino acid) were discovered in cattle, and the discovery of anserine (a methylated product of carnosine; ß-alanyl-1-methyl-L-histidine) also originated with cattle. These five nutrients are highly abundant in beef, and have important physiological roles in anti-oxidative and anti-inflammatory reactions, as well as neurological, muscular, retinal, immunological and cardiovascular function. Of particular note, taurine, carnosine, anserine, and creatine are absent from plants, and hydroxyproline is negligible in many plant-source foods. Consumption of 30 g dry beef can fully meet daily physiological needs of the healthy 70-kg adult human for taurine and carnosine, and can also provide large amounts of creatine, anserine and 4-hydroxyproline to improve human nutrition and health, including metabolic, retinal, immunological, muscular, cartilage, neurological, and cardiovascular health. The present review provides the public with the much-needed knowledge of nutritionally and physiologically significant amino acids, dipeptides and creatine in animal-source foods (including beef). Dietary taurine, creatine, carnosine, anserine and 4-hydroxyproline are beneficial for preventing and treating obesity, cardiovascular dysfunction, and ageing-related disorders, as well as inhibiting tumorigenesis, improving skin and bone health, ameliorating neurological abnormalities, and promoting well being in infants, children and adults. Furthermore, these nutrients may promote the immunological defense of humans against infections by bacteria, fungi, parasites, and viruses (including coronavirus) through enhancing the metabolism and functions of monocytes, macrophages, and other cells of the immune system. Red meat (including beef) is a functional food for optimizing human growth, development and health.


Assuntos
Anserina/metabolismo , Carnosina/metabolismo , Creatina/metabolismo , Hidroxiprolina/metabolismo , Carne/análise , Taurina/metabolismo , Animais , Bovinos , Humanos , Valor Nutritivo
11.
Carbohydr Polym ; 233: 115856, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32059907

RESUMO

A water-soluble polysaccharide (SPAW) was purified from Safflower and it was identified to be (1→3)-linked ß-d-Glucan. The therapeutic effect and underlying mechanism of SPAW on steroid-induced avascular necrosis of the femoral head (SANFH) in a rabbit model was performed here. The abnormal histopathologic changes and apoptosis of femoral head in model group were significantly reverted after SPAW (25, 100 and 200 mg/kg) administration for 60 days, as evidenced by the a decline of empty lacunae rate, the average bone marrow fat cell size and the proportion of apoptotic cells. Furthermore, administration of SPAW significantly decreased the Bax and caspase-3 protein expression, but increased the protein expression of Bcl-2 when compared these in model rabbits. Meanwhile, increased hydroxyproline (HOP) and decreased serum hexosamine (HOM) concentration in rabbit serum were turned to the opposite way. The present study suggested that SPAW may provide an alternative treatment for the treatment of SANFH.


Assuntos
Carthamus tinctorius/química , Necrose da Cabeça do Fêmur/tratamento farmacológico , Glucanos/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Feminino , Cabeça do Fêmur/patologia , Necrose da Cabeça do Fêmur/induzido quimicamente , Glucanos/química , Glucanos/isolamento & purificação , Hexosaminas/metabolismo , Cavalos , Hidroxiprolina/metabolismo , Masculino , Hemissuccinato de Metilprednisolona , Peso Molecular , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Coelhos , Ratos , Proteína X Associada a bcl-2/metabolismo
12.
Biomed Res Int ; 2020: 5052028, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32090096

RESUMO

Achilles tendinitis caused by overuse, aging, or gradual wear induces pain, swelling, and stiffness of Achilles tendon and leads to tendon rupture. This study was performed to investigate the suppression of inflammation responses in interleukin-1ß- (IL-1ß-) stimulated tenocytes in vitro and the suppression of the progression of Achilles tendinitis-induced rat models in vivo using dexamethasone-containing porous microspheres (DEX/PMSs) for a sustained intratendinous DEX delivery. DEX from DEX/PMSs showed the sustained release of DEX. Treatment of IL-1ß-stimulated tenocytes with DEX/PMSs suppressed the mRNA levels for COX-2, IL-1ß, IL-6, and TNF-α. The intratendinous injection of DEX/PMSs into Achilles tendinitis rats both decreased the mRNA levels for these cytokines and increased mRNA levels for anti-inflammatory cytokines IL-4 and IL-10 in tendon tissues. Furthermore, DEX/PMSs effectively prevented tendon degeneration by enhancing the collagen content and biomechanical properties. Our findings suggest that DEX/PMSs show great potential as a sustained intratendinous delivery system for ameliorating inflammation responses as well as tendon degeneration in Achilles tendinitis.


Assuntos
Tendão do Calcâneo/patologia , Dexametasona/administração & dosagem , Dexametasona/uso terapêutico , Inflamação/tratamento farmacológico , Microesferas , Tendinopatia/tratamento farmacológico , Tendão do Calcâneo/efeitos dos fármacos , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Morte Celular/efeitos dos fármacos , Citocinas/genética , Citocinas/metabolismo , Dexametasona/farmacologia , Modelos Animais de Doenças , Liberação Controlada de Fármacos , Hidroxiprolina/metabolismo , Inflamação/complicações , Mediadores da Inflamação/metabolismo , Masculino , Porosidade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Suínos , Tendinopatia/complicações , Resistência à Tração , Resultado do Tratamento
13.
Life Sci ; 242: 117175, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31843528

RESUMO

AIMS: Ursodeoxycholic acid (UDCA) has been widely used in the treatment of primary biliary cholangitis (PBC) with chronic liver fibrosis, but its detailed mechanism remains unclear. This study was aimed to determine whether autophagic signaling is involved in the therapeutic effect of UDCA on liver fibrosis. METHODS: By using hepatic stellate cell (HSC) line LX2 and CCl4-induced fibrotic rat model, autophagy signaling was investigated by western blotting and mRFP-EGFP-LC3 tandem fluorescent tagged plasmid (ptfLC3) transfection technique. Anti-fibrotic profile was determined by western blotting, qRT-PCR, MTT assay, trypan blue, hydroxyproline assay and Masson staining. KEY FINDINGS: TGFß1 treatment decreased P62 accumulation and increased both autophagosomes and autolysosomes in LX2 cells, thereby elevated autophagic flux. Hydroxychloroquine (HCQ), antagonist of autophagy, was found to dramatically inhibit COL1A2 mRNA expression and cell proliferation in a dose-dependent manner. This coincides with the effect of UDCA intervention on collagen aggradation and cell viability. Meanwhile, UDCA inhibited TGFß1-induced autophagy flux. And rapamycin, agonist of autophagy, was found to impair the anti-fibrotic effect of UDCA. Moreover, study in vivo showed that UDCA alone or in combination with HCQ restored the CCl4-induced liver fibrosis in rodent models with autophagy inhibited profile. SIGNIFICANCE: Taken together, our study revealed that UDCA displays anti-fibrotic role by protecting HSC against production of collagen and inhibiting cellular viability involving autophagy inhibition and provide a new insight into the pharmacological basis of UDCA treatment for hepatic fibrosis.


Assuntos
Autofagia/efeitos dos fármacos , Cirrose Hepática/tratamento farmacológico , Ácido Ursodesoxicólico/uso terapêutico , Animais , Western Blotting , Linhagem Celular , Modelos Animais de Doenças , Humanos , Hidroxicloroquina/farmacologia , Hidroxiprolina/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Crescimento Transformador beta1/farmacologia
14.
Biochim Biophys Acta Mol Basis Dis ; 1866(3): 165633, 2020 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-31821850

RESUMO

The major clinical manifestation of the Primary Hyperoxalurias (PH) is increased production of oxalate, as a consequence of genetic mutations that lead to aberrant glyoxylate and hydroxyproline metabolism. Hyperoxaluria can lead to the formation of calcium-oxalate kidney stones, nephrocalcinosis and renal failure. Current therapeutic approaches rely on organ transplants and more recently modifying the pathway of oxalate synthesis using siRNA therapy. We have recently reported that the metabolism of trans-4-hydroxy-L-proline (Hyp), an amino acid derived predominantly from collagen metabolism, is a significant source of oxalate production in individuals with PH2 and PH3. Thus, the first enzyme in the Hyp degradation pathway, hydroxyproline dehydrogenase (HYPDH), represents a promising therapeutic target for reducing endogenous oxalate production in these individuals. This is supported by the observation that individuals with inherited mutations in HYPDH (PRODH2 gene) have no pathological consequences. The creation of mouse models that do not express HYPDH will facilitate research evaluating HYPDH as a target. We describe the phenotype of the Prodh2 knock out mouse model and show that the lack of HYPDH in PH mouse models results in lower levels of urinary oxalate excretion, consistent with our previous metabolic tracer and siRNA-based knockdown studies. The double knockout mouse, Grhpr KO (PH2 model) and Prodh2 KO, prevented calcium-oxalate crystal deposition in the kidney, when placed on a 1% Hyp diet. These observations support the use of the Grhpr KO mice to screen HYPDH inhibitors in vivo. Altogether these data support HYPDH as an attractive therapeutic target for PH2 and PH3 patients.


Assuntos
Glicolatos/metabolismo , Glicolatos/urina , Hidroxiprolina/metabolismo , Hiperoxalúria Primária/metabolismo , Oxalatos/metabolismo , Oxalatos/urina , Oxirredutases/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Cálcio/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Rim/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Prolina Oxidase/metabolismo
15.
Life Sci ; 240: 117096, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31760097

RESUMO

Aim Liver fibrosis represents a massive global health burden with limited therapeutic options. Thus, the need for curative options is evident. Thus, this study aimed to assess the potential antifibrotic effect of honokiol in Concanavalin A (Con A) induced immunological model of liver fibrosis as well the possible underlying molecular mechanisms. METHODS: Male Sprague-Dawley rats were treated with either Con A (20 mg/kg, IV) and/or honokiol (10 mg/kg, orally) for 4 weeks. Hepatotoxicity indices were as well as histopathological evaluation was done. Hepatic fibrosis was assessed by measuring alpha smooth muscle actin (α-SMA) expression and collagen fibers deposition by Masson's trichrome stain and hydroxyproline content. To elucidate the underlying molecular mechanisms, the effect of honokiol on oxidative stress, inflammatory markers as well as transforming growth factor beta (TGF-ß)/SMAD and mitogen-activated protein kinase (MAPK) pathways was assessed. KEY FINDINGS: Honokiol effectively reversed the hepatotoxicity indices elevations and abnormal histopathological changes induced by Con A. Besides, honokiol attenuated Con A-induced liver fibrosis by down-regulation of hydroxyproline levels, α-SMA expression together with a marked decrease in collagen fibers deposition. Mechanistically Con A induced oxidative stress, provocation of inflammatory responses and activation of TGF-ß/SMAD/MAPK pathways. Contrariwise, honokiol co-treatment significantly restored antioxidant defence mechanisms, down-regulated inflammatory cascades and inhibited TGF-ß/SMAD/MAPK signaling pathways. CONCLUSION: The results provide an evidence for the promising antifibrotic effect of honokiol that could be partially due to suppressing oxidative stress and inflammatory processes as well as inhibition of TGF-ß/SMAD/MAPK signaling pathways.


Assuntos
Compostos de Bifenilo/uso terapêutico , Lignanas/uso terapêutico , Cirrose Hepática/prevenção & controle , Proteínas Quinases Ativadas por Mitógeno/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Proteínas Smad/efeitos dos fármacos , Fator de Crescimento Transformador beta/efeitos dos fármacos , Actinas/metabolismo , Animais , Concanavalina A , Hidroxiprolina/metabolismo , Inflamação/tratamento farmacológico , Inflamação/patologia , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/patologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Análise de Sobrevida
16.
Eur J Pharm Sci ; 141: 105042, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31634554

RESUMO

Skin aging affects personal image and health. α - lipoic acid (ALA), with excellent free radical scavenging capacity, was used in this research to prepare W/O emulsion. Considering the instability of ALA, ionic liquid strategy was adopted to heighten the solubility of ALA for dissolving in water phase. The mechanism of different ionic liquids (ILs) on skin retention of ALA was investigated by in vitro skin permeation experiment, emulsion quality characterization, rheological test, ATR - FTIR and molecular simulation. The results showed that ionic liquid strategy had a positive influence on the solubilization of ALA. Different ILs were different in skin retention and regulated by skin layers rather than drug release, in which ALA - triethanolamine (ALA - TEOA) presented the best affinity with both stratum corneum (SC) and viable epidermis and dermis (VED), while ALA - N - (2 - Hydroxyethyl) piperidine (ALA - HEPP) as well as ALA - N - (2 - hydroxyethyl) pyrrolidine (ALA - HEPR) showed affinity with either SC or VED respectively. Finally, the emulsion presented brilliant anti - aging efficacy. This study provided a new method of emulsion research and had great significance for the development of topical formulations.


Assuntos
Líquidos Iônicos/administração & dosagem , Envelhecimento da Pele/efeitos dos fármacos , Ácido Tióctico/administração & dosagem , Administração Cutânea , Animais , Emulsões , Hidroxiprolina/metabolismo , Líquidos Iônicos/química , Masculino , Ratos Wistar , Pele/efeitos dos fármacos , Pele/metabolismo , Pele/efeitos da radiação , Absorção Cutânea/efeitos dos fármacos , Envelhecimento da Pele/efeitos da radiação , Ácido Tióctico/química , Raios Ultravioleta/efeitos adversos
17.
Int J Biol Macromol ; 144: 781-790, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31678100

RESUMO

The present study investigates the purification, structure and in vitro antioxidant activities of a novel water soluble polysaccharide (LWSP) extracted from Linum usitatissimum L. seeds and evaluates the in vivo wound healing performance on CO2 laser fractional burn in a rat model. LWSP is a heteropolysaccharide that consists of glucose, mannose, xylose and arabinose. Three different tests were used to evaluate the antioxidant activity of this carbohydrate. The scavenging activity against 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical at a concentration of 5 mg/ml was 99.77%. The total antioxidant capacity of LWSP at 12 mg/ml was equivalent to 166.61 µg acsorbic acid. LWSP displayed a high protection effect against DNA damage induced byhydroxyl radical. No hemolytic activity was observed towards human erythrocytes. LWSP was tested in functional properties. The results showed good emulsion properties and high water (WHC) and oil holding (OHC) capacities (11.23 and 1.05%, respectively). In addition, the application of LWSP on the burn wound site in rat model increased significantly the percentage of burn contraction (98.6%) after 8 days of injury. According to the histological assessment, the LWSP-treated group had a higher content of hydroxyproline (846. 67 ±â€¯92.28 mg/g tissue) than the other groups. Overall, the findings demonstrate that the application of this novel LWSP may open promising opportunities for burn wound healing in modern medicine.


Assuntos
Antioxidantes/química , Queimaduras/tratamento farmacológico , Linho/química , Extratos Vegetais/química , Polissacarídeos/química , Sementes/química , Cicatrização/efeitos dos fármacos , Animais , Antioxidantes/farmacologia , Benzotiazóis/química , Linhagem Celular , Emulsões/química , Eritrócitos/citologia , Sequestradores de Radicais Livres/metabolismo , Humanos , Interações Hidrofóbicas e Hidrofílicas , Hidroxiprolina/metabolismo , Lasers , Masculino , Extratos Vegetais/farmacologia , Polissacarídeos/farmacologia , Ratos Wistar , Ácidos Sulfônicos/química
18.
Int J Biol Macromol ; 144: 954-966, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31672634

RESUMO

This work was conducted to evaluate the compatibility between physicochemical, antioxidant and morphological properties of polysaccharide (FRP) extracted from red marine alga Falkenbergia rufolanosa reinforced by poly (vinyl alcohol) (PVA) composed films at different ratios of FRP/PVA: F1 (70:30), F2 (50:50), F3 (30:70) and PVA (100% PVA) and the potential wound healing effects. As assessed, FRP/PVA prepared films were heterogeneous, slightly opaque with a rough surface as ascertained by Fourier transform infrared spectroscopy, scanning electron microscopy and colorimetric parameters. Even, X-ray diffraction and glass transition results revealed a semi-crystalline structure of FRP composed films which decreased with increasing PVA ratios. The antioxidant activities of composite films depicted that F1 exhibited the highest antioxidant activity in vitro. Therefore, F1 was found to promote significantly the wound healing, after eight days of treatment, evidenced by higher wound appearance scores and a higher content of collagen (885.12 ±â€¯20.35 mg/g of tissue) confirmed by histological examination, when compared with control, CYTOL BASIC® and PVA-treated groups. All together, the marine-derived polysaccharide gave a substantial pledge for the development of biodegradable films as a potent antioxidant material and a promising agent for tissue regeneration.


Assuntos
Queimaduras/fisiopatologia , Lasers/efeitos adversos , Polissacarídeos/química , Polissacarídeos/farmacologia , Álcool de Polivinil/química , Rodófitas/química , Cicatrização/efeitos dos fármacos , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Queimaduras/etiologia , Queimaduras/metabolismo , Queimaduras/patologia , Colágeno/metabolismo , Feminino , Hidroxiprolina/metabolismo , Polissacarídeos/uso terapêutico , Ratos , Ratos Wistar , Solubilidade
19.
Biomech Model Mechanobiol ; 19(1): 99-112, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31270728

RESUMO

Deposition of elastin and collagen in the aorta correlates with increases in blood pressure and flow during development, suggesting that the aorta adjusts its mechanical properties in response to hemodynamic stresses. Elastin knockout (Eln-/-) mice have high blood pressure and pathological remodeling of the aorta and die soon after birth. We hypothesized that decreasing blood pressure in Eln-/- mice during development may reduce hemodynamic stresses and alleviate pathological remodeling of the aorta. We treated Eln+/+ and Eln-/- mice with the anti-hypertensive medication captopril throughout embryonic development and then evaluated left ventricular (LV) pressure and aortic remodeling at birth. We found that captopril treatment decreased Eln-/- LV pressure to values near Eln+/+ mice and alleviated the wall thickening and changes in mechanical behavior observed in untreated Eln-/- aorta. The changes in thickness and mechanical behavior in captopril-treated Eln-/- aorta were not due to alterations in measured elastin or collagen amounts, but may have been caused by alterations in smooth muscle cell (SMC) properties. We used a constitutive model to understand how changes in stress contributions of each wall component could explain the observed changes in composite mechanical behavior. Our modeling results show that alterations in the collagen natural configuration and SMC properties in the absence of elastin may explain untreated Eln-/- aortic behavior and that partial rescue of the SMC properties may account for captopril-treated Eln-/- aortic behavior.


Assuntos
Aorta/crescimento & desenvolvimento , Captopril/farmacologia , Elastina/deficiência , Estresse Mecânico , Remodelação Vascular/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Aorta/efeitos dos fármacos , Fenômenos Biomecânicos/efeitos dos fármacos , Pressão Sanguínea/efeitos dos fármacos , Desmosina/metabolismo , Elastina/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Frequência Cardíaca/efeitos dos fármacos , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/fisiopatologia , Hidroxiprolina/metabolismo , Camundongos Knockout , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Angiotensina/genética , Receptores de Angiotensina/metabolismo
20.
Sci Rep ; 9(1): 16875, 2019 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-31728030

RESUMO

The higher-order architecture observed in biological systems, like viruses, is very effective in nucleic acid transport. The replications of this system has been attempted with both synthetic and naturally occurring polymers with mixed results. Here we describe a peptide/siRNA quaternary complex that functions as an siRNA delivery system. The rational design of a peptide assembly is inspired by the viral capsids, but not derived from them. We selected the collagen peptide (COL) to provide the structural stability and the folding framework, and hybridize it with the cell penetrating peptide (CPP) that allows for effective penetration of biological barriers. The peptide/siRNA quaternary complex forms stoichiometric, 10 nm nanoparticles, that show fast cellular uptake (<30 min), effective siRNA release, and gene silencing. The complex provides capsid-like protection for siRNA against nucleases without being immunostimulatory, or cytotoxic. Our data suggests that delivery vehicles based on synthetic quaternary structures that exhibit higher-order architecture may be effective in improving delivery and release of nucleic acid cargo.


Assuntos
Peptídeos Penetradores de Células/metabolismo , Colágeno/metabolismo , Inativação Gênica , Técnicas de Transferência de Genes , Polímeros/metabolismo , RNA Interferente Pequeno/genética , Animais , Transporte Biológico , Materiais Biomiméticos/química , Materiais Biomiméticos/metabolismo , Capsídeo/química , Carbocianinas/química , Carbocianinas/metabolismo , Peptídeos Penetradores de Células/química , Colágeno/química , Fluoresceína-5-Isotiocianato/química , Fluoresceína-5-Isotiocianato/metabolismo , Corantes Fluorescentes/química , Corantes Fluorescentes/metabolismo , Genes Reporter , Proteínas de Fluorescência Verde/antagonistas & inibidores , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Hidroxiprolina/química , Hidroxiprolina/metabolismo , Camundongos , Conformação Molecular , Células NIH 3T3 , Nanopartículas/química , Polímeros/química , RNA Interferente Pequeno/metabolismo
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