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1.
Talanta ; 209: 120487, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31892008

RESUMO

When drug nanocarriers enter a physiological environment, their surface gets coated by a dynamic biomolecular corona (BMC) mainly constituted by proteins. Although a deep investigation has been performed on the composition of BMC in terms of proteins, scarce attention has been posed to low molecular weight metabolites present in human plasma. In this work, for the first time, the investigation of the BMC of liposomal nanoparticles (NPs) constituted by 1,2-dioleoyl-3-trimethylammonium-propane polar lipid has been carried out by an ultra-high performance liquid chromatography coupled to high-resolution mass spectrometry based untargeted metabolomics approach. Compounds were tentatively identified based on matches with online databases and comparison of MS/MS spectra with available spectral libraries. Moreover, a comparison of three metabolite extraction strategies, including an ultrafiltration membrane based protocol, a methanol extraction based protocol, and Wessel & Flügge protocol, was performed. Methanol extraction procedure resulted in the widest metabolic coverage of liposomal NP BMC. A total of 193 metabolites has been tentatively identified, 166 of which belonged to the class of lipids including phospholipids, steroids, carnitines, fatty alcohols, diglycerides and fatty acids. The high abundance of lipids in the BMC can be explained by the adsorption of plasma lipoproteins onto liposome surface, confirming previous works on other kinds of NPs. Lipids are important bioactive molecules, which could impact NP circulation and uptake by cells. Extending the investigation of BMC beyond the protein corona and towards the "lipid corona" may be the keystone of a better understanding and control of NP fate in human body.


Assuntos
Lipídeos/sangue , Lipossomos/química , Cromatografia Líquida de Alta Pressão , Ácidos Graxos Monoinsaturados/química , Humanos , Lipidômica/métodos , Espectrometria de Massas , Metaboloma , Nanopartículas/química , Compostos de Amônio Quaternário/química
2.
Drug Des Devel Ther ; 13: 4195-4205, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31849451

RESUMO

Background: Cancer is one of the chronic health conditions worldwide. Various therapeutically active compounds from medicinal plants were the current focus of this research in order to uncover a treatment regimen for cancer. Anchusa arvensis (A. anchusa) (L.) M.Bieb. contains many biologically active compounds. Methods: In the current study, new ester 3-hydroxyoctyl -5- trans-docosenoate (compound-1) was isolated from the chloroform soluble fraction of A. anchusa using column chromatography. Using MTT assay, the anticancer effect of the compound was determined in human hepatocellular carcinoma cells (HepG-2) compared with normal epithelial cell line (Vero). DPPH and ABTS radical scavenging assays were performed to assess the antioxidant potential. The Molecular Operating Environment (MOE-2016) tool was used against tyrosine kinase. Results: The structure of the compound was elucidated based on IR, EI, and NMR spectroscopy technique. It exhibited a considerable cytotoxic effect against HepG-2 cell lines with IC50 value of 6.50 ± 0.70 µg/mL in comparison to positive control (doxorubicin) which showed IC50 value of 1.3±0.21 µg/mL. The compound did not show a cytotoxic effect against normal epithelial cell line (Vero). The compound also exhibited significant DPHH scavenging ability with IC50 value of 12 ± 0.80 µg/mL, whereas ascorbic acid, used as positive control, demonstrated activity with IC50 = 05 ± 0.15 µg/mL. Similarly, it showed ABTS radical scavenging ability (IC50 = 130 ± 0.20 µg/mL) compared with the value obtained for ascorbic acid (06 ± 0.85 µg/mL). In docking studies using MOE-2016 tool, it was observed that compound-1 was highly bound to tyrosine kinase by having two hydrogen bonds at the hinge region. This good bonding network by the compound might be one of the reasons for showing significant activity against this enzyme. Conclusion: Our findings led to the isolation of a new compound from A. anchusa which has significant cytotoxic activity against HepG-2 cell lines with marked antioxidant potential.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Benzotiazóis/antagonistas & inibidores , Compostos de Bifenilo/antagonistas & inibidores , Boraginaceae/química , Ésteres/farmacologia , Ácidos Graxos Monoinsaturados/farmacologia , Picratos/antagonistas & inibidores , Ácidos Sulfônicos/antagonistas & inibidores , Animais , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Proliferação de Células/efeitos dos fármacos , Chlorocebus aethiops , Simulação por Computador , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Ésteres/química , Ésteres/isolamento & purificação , Ácidos Graxos Monoinsaturados/química , Ácidos Graxos Monoinsaturados/isolamento & purificação , Células Hep G2 , Humanos , Simulação de Acoplamento Molecular , Estrutura Molecular , Plantas Medicinais , Relação Estrutura-Atividade , Células Vero
4.
Int J Nanomedicine ; 14: 6575-6585, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31616144

RESUMO

Background and purpose: In a past study, we developed and optimized a novel cationic PEGylated niosome containing anticancer drugs (doxorubicin or quercetin) and siRNA. This study intended to evaluate the anti-tumor effects of the combination therapy to target both the proteins and genes responsible for the development of gastric cancer. CDC20, known as an oncogene, is a good potential therapeutic candidate for gastric cancer. Methods: In order to increase the loading capacity of siRNA and achieve appropriate physical properties, we optimized the cationic PEGylated niosome in terms of the amount of the cationic lipids. Drugs (doxorubicin and quercetin) and CDC20siRNA were loaded into the co-delivery system, and physical characteristics, thermosensitive controlled-release, gene silencing efficiency, and apoptosis rate were determined. Results: The results showed that the designed co-delivery system for the drugs and gene silencer had an appropriate size and a high positive charge for loading siRNA, and also showed a thermosensitive drug release behavior, which successfully silenced the CDC20 expression when compared with the single delivery of siRNA or the drug. Moreover, the co-delivery of drugs and CDC20siRNA exhibited a highly inhibitory property for the cell growth of gastric cancer cells. Conclusion: It seems that the novel cationic PEGylated niosomes co-loaded with anticancer drug and CDC20siRNA has a promising application for the treatment of gastric cancer.


Assuntos
Proteínas Cdc20/metabolismo , Doxorrubicina/uso terapêutico , Polietilenoglicóis/química , Quercetina/uso terapêutico , RNA Interferente Pequeno/metabolismo , Neoplasias Gástricas/tratamento farmacológico , Neoplasias Gástricas/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Cátions , Proteínas Cdc20/genética , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Liberação Controlada de Fármacos , Endocitose , Ácidos Graxos Monoinsaturados/química , Inativação Gênica/efeitos dos fármacos , Humanos , Lipossomos , Tamanho da Partícula , Compostos de Amônio Quaternário/química , Quercetina/farmacologia , RNA Interferente Pequeno/genética , Eletricidade Estática , Temperatura
5.
Environ Sci Pollut Res Int ; 26(31): 31685-31698, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31485936

RESUMO

Suspended sediments (SSs) were examined regarding the content of fatty acids (FAs) to associate them with sources of soil entry into the river. The source of organic matter was traced through fatty acid distribution, as well as erosion. Also, TOC, TN, and TOC/NT were used to support the results of FAs. For this, a tropical river was chosen to understand the main source of input considering the level of land occupation along the river. The Barigui river, in southern Brazil, was segmented in four distinct areas regarding the soil occupation (P1, P2, P3, and P4). Nine sampling campaigns were conducted from Nov/2014 to Nov/2015 using a time-integrated sampler. Site P1 has the lowest level of urbanization and showed the lowest concentration of FAs (16.35 µg-1). In contrast, site P4, the most urbanized, showed the highest content of fatty acids, including those associated with erosion, 378.53 µg g-1, specifically those with long chains. The mean concentrations of the saturated fatty acids (FAs) was 283.40 µg g-1, monounsaturated fatty acids (MUFAs) was 79.46 µg g-1, and polyunsaturated fatty acids (PUFAs) was 15.66 µg g-1. Twenty-seven fatty acids were examined, nevertheless C15:0, C16:0, C18:0, and C18:1ω9 prevailed in all samples. Generally, those acids indicate sewage inputs. Statics analyses were used to find the relation between the source of organic matter (autochthonous, allochthones, and anthropogenic) and FAs. Finally, the input of organic matter is associated with land occupation, which can be distinguished by FA distribution.


Assuntos
Ácidos Graxos Monoinsaturados/química , Ácidos Graxos/análise , Esgotos/análise , Brasil , Ácidos Graxos/química , Rios , Urbanização
6.
DNA Cell Biol ; 38(10): 1048-1055, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31433200

RESUMO

DNA condensed agents can improve the transfection efficiency of the cationic liposome delivery system. However, various condensed agents have distinct transfection efficiency and cellular cytotoxicity. The object of this study was to screen the optimal agents with the high transfection efficiency and low cytotoxicity from four polymer compressive materials, polyethylenimine (PEI), chitosan, poly-l-lysine (PLL), and spermidine. DNA was precompressed with these four agents and then combined to cationic liposomes. Subsequently, the entrapment and transfection efficiency of the obtained complexes were investigated. Finally, the particle sizes, cytotoxicity, and endocytosis fashion of these copolymers (Lipo-PEI, Lipo-chitosan, Lipo-PLL, and Lipo-spermidine) were examined. It was found that these four copolymers had significantly lower cytotoxicity and higher transfection efficiency (45.5%, 42.4%, 36.8%, and 47.4%, respectively) than those in the control groups. The transfection efficiency of Lipo-PEI and Lipo-spermidine copolymers were better than the other two copolymers. In 293T cells, nystatin significantly inhibited the transfection efficiency of Lipo-PEI-DNA and Lipo-spermidine-DNA (51.88% and 46.05%, respectively), which suggest that the endocytosis pathway of Lipo-spermidine and Lipo-PEI copolymers was probably caveolin dependent. Our study indicated that these dual-degradable copolymers especially liposome-spermidine copolymer could be used as the potential biocompatible gene delivery carriers.


Assuntos
Quitosana/química , Lipossomos/química , Polietilenoimina/química , Polilisina/química , Espermidina/química , Transfecção/métodos , Cátions , Caveolina 1/genética , Caveolina 1/metabolismo , Quitosana/metabolismo , Colesterol/química , Colesterol/metabolismo , Endocitose/efeitos dos fármacos , Endocitose/genética , Ácidos Graxos Monoinsaturados/química , Ácidos Graxos Monoinsaturados/metabolismo , Células HEK293 , Humanos , Lipossomos/metabolismo , Nistatina/farmacologia , Tamanho da Partícula , Plasmídeos/química , Plasmídeos/metabolismo , Polietilenoimina/metabolismo , Polilisina/metabolismo , Compostos de Amônio Quaternário/química , Compostos de Amônio Quaternário/metabolismo , Espermidina/metabolismo
7.
Cells ; 8(8)2019 07 29.
Artigo em Inglês | MEDLINE | ID: mdl-31362382

RESUMO

The dysfunction of oligodendrocytes (OLs) is regarded as one of the major causes of inefficient remyelination in multiple sclerosis, resulting gradually in disease progression. Oligodendrocytes are derived from oligodendrocyte progenitor cells (OPCs), which populate the adult central nervous system, but their physiological capability to myelin synthesis is limited. The low intake of essential lipids for sphingomyelin synthesis in the human diet may account for increased demyelination and the reduced efficiency of the remyelination process. In our study on lipid profiling in an experimental autoimmune encephalomyelitis brain, we revealed that during acute inflammation, nervonic acid synthesis is silenced, which is the effect of shifting the lipid metabolism pathway of common substrates into proinflammatory arachidonic acid production. In the experiments on the human model of maturating oligodendrocyte precursor cells (hOPCs) in vitro, we demonstrated that fish oil mixture (FOM) affected the function of hOPCs, resulting in the improved synthesis of myelin basic protein, myelin oligodendrocyte glycoprotein, and proteolipid protein, as well as sphingomyelin. Additionally, FOM reduces proinflammatory cytokines and chemokines, and enhances fibroblast growth factor 2 (FGF2) and vascular endothelial growth factor (VEGF) synthesis by hOPCs was also demonstrated. Based on these observations, we propose that the intake of FOM rich in the nervonic acid ester may improve OL function, affecting OPC maturation and limiting inflammation.


Assuntos
Ácidos Graxos Monoinsaturados/farmacologia , Bainha de Mielina/metabolismo , Oligodendroglia/efeitos dos fármacos , Oligodendroglia/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Encefalomielite Autoimune Experimental , Ésteres , Ácidos Graxos Monoinsaturados/química , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Lipídeos , Metabolômica/métodos , Camundongos , Estrutura Molecular , Células-Tronco Neurais/citologia , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/metabolismo
8.
Int J Nanomedicine ; 14: 4353-4366, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31354265

RESUMO

Purpose: Gene therapy has become a promising remedy to treat disease by modifying the person's genes. The therapeutic potential of related tools such as CRISPR-Cas9 depends on the efficiency of delivery to the targeted cells. Numerous transfection reagents have been designed and lots of efforts have been devoted to develop carriers for this purpose. Therefore, the aim of the present study was to develop novel cholesterol-rich lipid-based nanoparticles to enhance transfection efficiency and serum stability. Materials and methods: We constructed two-, three- and four-component cationic liposomes (CLs) to evaluate the combined effect of cholesterol domain and DOPE (dioleoyl phosphatidylethanolamine), a fusogenic lipid, and the PEG (polyethylene glycol) moiety location inside or outside of the cholesterol domain on transfection efficiency and other properties of the particle. Lipoplex formation and pDNA (plasmid DNA) entrapment were assessed by gel retardation assay at different N/P ratios (3, 5, 7). Physicochemical characteristics, cytotoxicity, serum stability and endosomal escape capability of the lipoplexes were studied and transfection potential was measured by firefly luciferase assay. Next, HEK293 cell line stably expressing GFP was utilized to demonstrate the editing of a reporter through Cas9 and sgRNA plasmids delivery by the selected CL formula, which showed the highest transfection efficiency. Results: Among the designed CLs, the four-component formula [DOTAP (1,2-dioleoyl-3-trimethylammoniumpropane)/DOPE/cholesterol/Chol-PEG (cholesterol-polyethylene glycol)] showed the highest rate of transfection at N/P 3. Finally, transfection of Cas9/sgRNA by this formulation at N/P 3 resulted in 39% gene-editing efficiency to knockout GFP reporter. The results also show that this CL with no cytotoxicity effect can totally protect the plasmids from enzymatic degradation in serum. Conclusion: The novel PEGylated cholesterol domain lipoplex providing serum stability, higher transfection efficiency and endosomal release can be used for in vivo Cas9/sgRNA delivery and other future gene-therapy applications.


Assuntos
Proteína 9 Associada à CRISPR/metabolismo , Sistemas CRISPR-Cas/genética , Colesterol/química , Edição de Genes , Nanopartículas/química , Transfecção/métodos , Cátions/química , Morte Celular , Colesterol/análogos & derivados , Ensaio de Desvio de Mobilidade Eletroforética , Endossomos/metabolismo , Ácidos Graxos Monoinsaturados/química , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Lipossomos/química , Tamanho da Partícula , Fosfatidiletanolaminas/química , Plasmídeos/metabolismo , Polietilenoglicóis/química , Compostos de Amônio Quaternário/química , RNA Guia/metabolismo , Eletricidade Estática
9.
Nanotechnology ; 30(41): 415604, 2019 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-31295734

RESUMO

The development of polymer-based nanoparticulate delivery systems for siRNA is important for the clinical success of gene therapy. However, there are some major drawbacks that need to be overcome. Short interfering RNA (siRNA) has been investigated as a potential therapeutic drug to silence disease-associated genes, but its usage is limited due to the lack of effective and safe nanocarriers. In this study, DOPE-PEI, a nanoparticle consisting of the fusogenic lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) conjugated with low-molecular-weight, 600 Da, branched polyethylenimine (PEI) was produced and optimized for siRNA delivery. This delivery system was modified with other components such as 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)2000] (DOPE-PEG2K), DOPE-PEG3.4K-bombesin and 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine/1,2-dioleoyl-3-trimethylammonium-propane (DOPE/DOTAP) and tested on PC-3 cells. The conjugation of DOPE to PEI polymer (DOPE-PEI) improved the efficiency of PEI to deliver siRNA into the cytosol and knockdown genes, but demonstrated high toxicity. The addition of DOPE-PEG2K reduced cellular toxicity by masking the surface positive charge of the DOPE-PEI/siRNA complex, with the incorporation of a gastrin-releasing peptide receptor (GRPR) targeting peptide and DOPE/DOTAP components improving the cellular uptake of siRNA into targeted cells and the siRNA knockdown efficiency.


Assuntos
Nanopartículas/química , Peptídeos/química , Polímeros/química , RNA Interferente Pequeno/administração & dosagem , Linhagem Celular Tumoral , Portadores de Fármacos/química , Ácidos Graxos Monoinsaturados/química , Técnicas de Silenciamento de Genes , Técnicas de Transferência de Genes , Terapia Genética/métodos , Humanos , Iminas/química , Lipídeos/química , Células PC-3 , Fosfatidiletanolaminas/química , Polietilenoglicóis/química , Polietilenos/química , Compostos de Amônio Quaternário/química , Receptores da Bombesina/metabolismo
10.
Gene Ther ; 26(9): 363-372, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31300730

RESUMO

Self-amplifying RNA (saRNA) is a promising biotherapeutic tool that has been used as a vaccine against both infectious diseases and cancer. saRNA has been shown to induce protein expression for up to 60 days and elicit immune responses with lower dosing than messenger RNA (mRNA). Because saRNA is a large (~9500 nt), negatively charged molecule, it requires a delivery vehicle for efficient cellular uptake and degradation protection. Lipid nanoparticles (LNPs) have been widely used for RNA formulations, where the prevailing paradigm is to encapsulate RNA within the particle, including the first FDA-approved small-interfering siRNA therapy. Here, we compared LNP formulations with cationic and ionizable lipids with saRNA either on the interior or exterior of the particle. We show that LNPs formulated with cationic lipids protect saRNA from RNAse degradation, even when it is adsorbed to the surface. Furthermore, cationic LNPs deliver saRNA equivalently to particles formulated with saRNA encapsulated in an ionizable lipid particle, both in vitro and in vivo. Finally, we show that cationic and ionizable LNP formulations induce equivalent antibodies against HIV-1 Env gp140 as a model antigen. These studies establish formulating saRNA on the surface of cationic LNPs as an alternative to the paradigm of encapsulating RNA.


Assuntos
Nanopartículas , RNA Mensageiro/administração & dosagem , Animais , Cátions , Ácidos Graxos Monoinsaturados/química , Feminino , Células HEK293 , Anticorpos Anti-HIV/biossíntese , Humanos , Lipídeos/administração & dosagem , Lipídeos/química , Luciferases de Vaga-Lume/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Nanopartículas/administração & dosagem , Nanopartículas/química , Tamanho da Partícula , Compostos de Amônio Quaternário/química , Estabilidade de RNA , RNA Mensageiro/imunologia , RNA Mensageiro/metabolismo , Ribonucleases/metabolismo , Transfecção , Produtos do Gene env do Vírus da Imunodeficiência Humana/imunologia
11.
Food Chem Toxicol ; 130 Suppl 1: 110622, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31238136

RESUMO

The existing information supports the use of this material as described in this safety assessment. Methyl 2-nonenoate was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, phototoxicity/photoallergenicity, skin sensitization, and environmental safety. Data from read-across analog ethyl trans-2,cis-4-decadienoate (CAS # 3025-30-7) show that methyl 2-nonenoate is not expected to be genotoxic. The repeated dose, reproductive, and local respiratory toxicity endpoints were evaluated using the TTC for a Cramer Class I material, and the exposure to methyl 2-nonenoate is below the TTC (0.03 mg/kg/day, 0.03 mg/kg/day, and 1.4 mg/day, respectively). Data from the target and read-across analog isobutyl-2-butenoate (CAS # 589-66-2) do not indicate the material is a sensitizer. The phototoxicity/photoallergenicity endpoints were evaluated based on data and UV spectra; methyl 2-nonenoate is not expected to be phototoxic/photoallergenic. The environmental endpoints were evaluated; methyl 2-nonenoate was found not to be PBT as per the IFRA Environmental Standards, and its risk quotients, based on its current volume of use in Europe and North America (i.e., PEC/PNEC), are <1.


Assuntos
Ácidos Graxos Monoinsaturados/química , Ácidos Graxos Monoinsaturados/toxicidade , Perfumes/química , Perfumes/toxicidade , Testes de Toxicidade/métodos , Animais , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Humanos , Estrutura Molecular , Medição de Risco
12.
Biomater Sci ; 7(8): 3158-3164, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31232421

RESUMO

Indocyanine green (ICG) is a clinically-approved near infrared (NIR) dye used for optical imaging. The dye is only slightly soluble in water and is prone to aggregation in saline solutions, so that alternative formulations can improve photophysical performance. Numerous nanoscale formulations of ICG have been described in the literature, but we sought to develop an approach that does not require additional purification steps. Pre-formed liposomes incorporating 45 mol% of the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) rapidly bind ICG, resulting in enhanced NIR optical properties. ICG binding is dependent on the amount of DOTAP incorporated in the liposomes. A dye-to-lipid mass ratio of [0.5 : 25] is sufficient for full complexation, without additional purification steps following mixing. NIR absorption, fluorescence intensity, and photoacoustic signals are increased for the liposome-bound dye. Not only is the optical character enhanced by simple mixing of ICG with liposomes, but retention in 4T1 mammary tumors is observed following intratumor injection, as assessed by fluorescence and photoacoustic imaging. Subsequent photothermal therapy with 808 nm laser irradiation is effective and results in tumor ablation without regrowth for at least 30 days. Thus, ICG optical properties and photothermal ablation outcomes can be improved by mixing the dye with pre-formed DOTAP liposomes in conditions that result in full dye-binding to the liposomes.


Assuntos
Técnicas de Ablação/métodos , Ácidos Graxos Monoinsaturados/química , Verde de Indocianina/química , Lipossomos/química , Neoplasias Mamárias Experimentais/terapia , Fenômenos Ópticos , Compostos de Amônio Quaternário/química , Animais , Feminino , Neoplasias Mamárias Experimentais/diagnóstico por imagem , Camundongos , Imagem Óptica , Fototerapia
13.
Lipids Health Dis ; 18(1): 145, 2019 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-31228942

RESUMO

The beneficial properties of fatty acids have been undervalued for several years. In contraposition, new studies reveal that fatty acids have an essential role for human health. The aim of the study is to demonstrate the clinical applications of fatty acids present in sea buckthorn oil. The composition of fatty acids found in sea buckthorn (Hippophae rhamnoides) oil is unique for this species, presenting a vast range of health benefits for humans and therefore it is highly valued by both biomedicine and the cosmetic industry. In this way, we will see the clinical effect of monounsaturated, polyunsaturated and saturated fatty acids that constitute sea buckthorn oil and how they contribute to the correct function of the organism. Despite there being studies that support the positive effects of sea buckthorn fatty acids, they are limited. Hence, most of the results obtained in this review are from studies of isolated fatty acids instead of fatty acids extracted from sea buckthorn oil. These facts permit to demonstrate the effect of sea buckthorn fatty acids separately but we lost the possibility of detecting a synergic effect of all of them. More studies are necessary to certify the clinical application of the fatty acids present in sea buckthorn oil as well as discovering possible synergies between them.


Assuntos
Ácidos Graxos/química , Hippophae/química , Osso e Ossos/efeitos dos fármacos , Olho/efeitos dos fármacos , Ácidos Graxos/farmacologia , Ácidos Graxos Monoinsaturados/química , Ácidos Graxos Monoinsaturados/farmacologia , Ácidos Graxos Ômega-3/química , Ácidos Graxos Ômega-3/farmacologia , Ácidos Graxos Ômega-6/química , Ácidos Graxos Ômega-6/farmacologia , Humanos , Pele/efeitos dos fármacos
14.
Nanomedicine (Lond) ; 14(9): 1153-1171, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31050581

RESUMO

Aim: To develop a peptide/phospholipid hybrid system for gastrin-releasing peptide receptor (GRPR)-targeted delivery of pDNA or siRNA. Materials & methods: A multifunctional GRPR-targeted peptide R9-K(GALA)-BBN(6-14) was combined with a phospholipid oligonucleotide delivery system (1:1 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine and 1,2-dioleoyl-3-trimethylammonium-propane) and evaluated for pDNA and siRNA delivery in terms of complex size, toxicity, receptor-targeted delivery and gene expression or knockdown efficiency. Results: By combining peptide and phospholipid delivery systems, synergistic improvements in gene expression and knockdown were observed when compared with either system alone. The optimized formulation demonstrated high levels of EGFP expression and EGFP knockdown, GRPR-targeted delivery, enhanced endosomal release and minimal toxicity. Conclusion: The peptide/phospholipid hybrid system provides efficient GRPR-targeted DNA/siRNA delivery.


Assuntos
DNA/administração & dosagem , Ácidos Graxos Monoinsaturados/química , Peptídeos/química , Fosfatidiletanolaminas/química , Compostos de Amônio Quaternário/química , RNA Interferente Pequeno/administração & dosagem , Receptores da Bombesina/metabolismo , Sobrevivência Celular , Técnicas de Silenciamento de Genes , Técnicas de Transferência de Genes , Proteínas de Fluorescência Verde/genética , Humanos , Terapia de Alvo Molecular , Células PC-3 , Plasmídeos , Transfecção
15.
Biomed Res Int ; 2019: 2640684, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31119160

RESUMO

The aim of the present study was to extract and characterize bioactive components from separate body organs of Holothuria leucospilota. Preliminary qualitative assessment of the crude extracts was positive for phenols, terpenoids, carbohydrates, flavonoids, saponins, glycosides, cardiac glycosides, steroids, phlobatannins, and tannins in all body organs evaluated. Phenolics were the most abundant group of bioactives accounting for approximately 80%. The extraction solvent mixtures that yielded most compounds evaluated were methanol/acetone (3:1, v:v) and methanol/distilled water (3:1, v:v). In other analyses, GC-MS data revealed diverse metabolic and biologically active compounds, where those in high concentrations included 2-Pentanone, 4-hydroxy-4-methyl- among the ketones; phenol- 2,4-bis(1,1-dimethylethyl)-, a phenol group; and 2-Chlorooctane, a hydrocarbon. Among FA and their methyl/ethyl esters, n-hexadecanoic acid, 5,8,11,14-eicosatetraenoic acid ethyl ester (arachidonic acid), and 5,8,11,14,17-eicosapentaenoic acid methyl ester (EPA) were among the most abundant FAMEs accounting for approximately 50% of the subgroups measured. Data from GC-FID analysis revealed methyl laurate (C12:0), methyl myristate (C14:0), methyl palmitate (C16:0), and methyl stearate (18:0) methyl esters as the most abundant saturated FA, whereas cis-9-oleic methyl ester (C18:1) and methyl linoleate (C18:2) were found as the major monounsaturated FA and PUFA FAMEs, respectively, in the body wall of the species. Taken together, the extraction and characterization of different categories of metabolically and biologically active compounds in various organ extracts of H. leucospilota suggest that the species is potentially a rich source of cholesterol-lowering, antioxidant, antimicrobial, and anticancer agents. These substances are known to benefit human health and assist in disease prevention. These findings justify the use of sea cucumbers in traditional folklore medication and the current interest and attention focused on the species to mine for bioactives in new drugs research.


Assuntos
Anti-Infecciosos/química , Antioxidantes/química , Ácidos Graxos Monoinsaturados/química , Pepinos-do-Mar/química , Animais , Anti-Infecciosos/uso terapêutico , Antioxidantes/uso terapêutico , Ácidos Graxos Monoinsaturados/uso terapêutico , Flavonoides/química , Cromatografia Gasosa-Espectrometria de Massas , Glicosídeos/química , Humanos , Saponinas/química , Terpenos/química
16.
Int J Nanomedicine ; 14: 2637-2653, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31043779

RESUMO

Background: For the past few years, gene-therapy has recently shown considerable clinical benefit in cancer therapy, and the applications of gene therapies in cancer treatments continue to increase perennially. EZH2, an ideal candidate for tumor gene therapy, plays an important role in the tumorigenesis. Methods: In this study, we developed a novel gene delivery system with a self-assembly method by Methoxy polyethylene glycol-polycaprolactone (MPEG-PCL) and DOTAP(DMC). And EZH2si-DMC was used to research anti-glioma both in vitro and in vivo. Results: DMC with zeta-potential value of 36.7 mV and size of 35.6 nm showed good performance in the delivery siRNA to glioma cell in vitro with high 98% transfection efficiency. EZH2si-DMC showed good anti-glioma effect in vitro through inducing cell apoptosis and inhibiting cell growth. What's more, treatment of tumor-bearing mice with DMC-EZH2si complex had significantly inhibited tumor growth at the subcutaneous model in vivo by inhibiting EZH2 protein expression, promoting apoptosis and reducing proliferation. Conclusion: The EZH2 siRNA and DMC complex may be used to treat the glioma in clinical as a new drug.


Assuntos
Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Glioma/tratamento farmacológico , Terapia de Alvo Molecular , Nanopartículas/química , RNA Interferente Pequeno/metabolismo , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Ácidos Graxos Monoinsaturados/síntese química , Ácidos Graxos Monoinsaturados/química , Feminino , Glioma/patologia , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Micelas , Nanopartículas/ultraestrutura , Poliésteres/síntese química , Poliésteres/química , Polietilenoglicóis/síntese química , Polietilenoglicóis/química , Compostos de Amônio Quaternário/síntese química , Compostos de Amônio Quaternário/química , RNA Interferente Pequeno/administração & dosagem
17.
J Immunol ; 202(12): 3524-3536, 2019 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-31053626

RESUMO

Certain types of cationic lipids have shown promise in cancer immunotherapy, but their mechanism of action is poorly understood. In this study, we describe the properties of an immunotherapeutic consisting of the pure cationic lipid enantiomer R-1,2-dioleoyl-3-trimethyl-ammonium-propane (R-DOTAP) formulated with modified viral or self-peptide Ags. R-DOTAP formulations with peptide Ags stimulate strong cross-presentation and potent CD8 T cell responses associated with a high frequency of polyfunctional CD8 T cells. In a human papillomavirus tumor model system, a single s.c. injection of tumor-bearing mice with R-DOTAP plus human papillomavirus Ags induces complete regression of large tumors associated with an influx of Ag-specific CD8 T cells and a reduction of the ratio of regulatory/Ag-specific CD8 T cells. R-DOTAP also synergizes with an anti-PD1 checkpoint inhibitor, resulting in a significant inhibition of B16 melanoma tumor growth. We found that R-DOTAP stimulates type I IFN production by dendritic cells in vivo and in vitro. s.c. injection of R-DOTAP results in an IFN-dependent increase in draining lymph node size and a concomitant increase in CD69 expression. Using knockout mice, we show that type I IFN is required for the induction of CD8 T cell activity following administration of R-DOTAP plus Ag. This response requires Myd88 but not TRIF or STING. We also show that R-DOTAP stimulates both TLR7 and 9. Collectively, these studies reveal that R-DOTAP stimulates endosomal TLRs, resulting in a Myd88-dependent production of type I IFN. When administered with Ag, this results in potent Ag-specific CD8 T cell responses and antitumor activity.


Assuntos
Imunoterapia Adotiva/métodos , Melanoma/terapia , Nanopartículas/metabolismo , Papillomaviridae/fisiologia , Infecções por Papillomavirus/terapia , Neoplasias Cutâneas/terapia , Linfócitos T Citotóxicos/imunologia , Animais , Células Cultivadas , Modelos Animais de Doenças , Ácidos Graxos Monoinsaturados/química , Humanos , Interferon Tipo I/metabolismo , Ativação Linfocitária , Melanoma/imunologia , Melanoma Experimental , Camundongos , Camundongos Endogâmicos C57BL , Camundongos SCID , Camundongos Transgênicos , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Nanopartículas/química , Infecções por Papillomavirus/imunologia , Compostos de Amônio Quaternário/química , Neoplasias Cutâneas/imunologia , Linfócitos T Citotóxicos/transplante
18.
Pharm Res ; 36(6): 81, 2019 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-30976936

RESUMO

PURPOSE: To develop cationic lipid-coated magnesium phosphate nanoparticles (LPP) for intracellular catalase (CAT) delivery. METHODS: Magnesium phosphate nanoparticles (MgP NP) were prepared by micro-emulsion precipitation and mixed with catalase-loaded cationic liposomes (DOTAP/cholesterol) to yield LPP formulation of catalase (LPP-CAT). The size and ζ-potential of LPP-CAT were measured by dynamic light scattering. The pH-sensitivity of LPP-CAT was determined by monitoring their degradation of hydrogen peroxide (H2O2) and their morphologies under transmission electron microscopy (TEM) at pH 7.4 and 5.5. The ability of LPP-CAT to protect MCF-7 cells against hydrogen peroxide was measured by MTS assay. ROS levels in EA.hy926 cells were measured after treatment with LPP-CAT. RESULTS: LPP-CAT were successfully prepared and carried an average diameter of <300 nm and ζ -potential of about +40 mV. At pH 5.5, LPP-CAT degraded H2O2 almost 4-fold as fast as pH 7.4 and displayed drastic morphological changes of an osmotic explosion. LPP-CAT protected MCF-7 cells from lethal level of exogenous H2O2 and significantly lowered the ROS levels in EA.hy926 cells. A lipid with a pH-sensitive conformational switch (flipid) further enhanced the protein delivery of LPP-CAT. CONCLUSION: LPP represents a promising nano-system for intracellular protein delivery.


Assuntos
Catalase/administração & dosagem , Portadores de Fármacos/química , Lipídeos/química , Compostos de Magnésio/química , Nanopartículas Metálicas/química , Fosfatos/química , Catalase/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Colesterol/química , Liberação Controlada de Fármacos , Ácidos Graxos Monoinsaturados/química , Humanos , Peróxido de Hidrogênio/metabolismo , Concentração de Íons de Hidrogênio , Lipossomos/química , Tamanho da Partícula , Compostos de Amônio Quaternário/química , Propriedades de Superfície
19.
J Agric Food Chem ; 67(16): 4425-4434, 2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-30945860

RESUMO

Grapholita molesta is a notorious fruit borer globally, causing severe damage to fruit production. To control the pest, one commonly used mean is pheromone-mediated management. As an important sex pheromone, Z-8-dodecenyl acetate (Z8-12: Ac), is often coformulated with other active ingredients to regulate the behavior of G. molesta. To uncover its interactions with G. molesta pheromone binding protein 2 (GmolPBP2) is used to help develop insect attractants. During 200 ns molecular dynamics simulations, two representative conformations of the GmolPBP2-Z8-12: Ac complex are selected. Conformation II at the time of 14-106 ns is dominantly maintained by the hydrophobic interactions and hydrogen bond. In Conformation I, which lasts from 106 to 200 ns, the hydrophobic interactions are enhanced while the hydrogen bond is quite weakened, due to the formation of a more sophisticated hydrophobic binding pocket and the enlargement of hydrogen bond distance. Taking the two conformations as a whole, the affinity between GmolPBP2 and Z8-12: Ac is crucially determined by three hot-spots including Phe11, Trp36, and Ile51. These results would provide a basis for the discovery, optimization, and design of leading compounds potentially active to attract G. molesta.


Assuntos
Proteínas de Transporte/química , Ácidos Graxos Monoinsaturados/química , Proteínas de Insetos/química , Mariposas/metabolismo , Feromônios/química , Sequência de Aminoácidos , Animais , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Ácidos Graxos Monoinsaturados/metabolismo , Feminino , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Cinética , Masculino , Dados de Sequência Molecular , Mariposas/química , Mariposas/genética , Feromônios/metabolismo , Ligação Proteica , Alinhamento de Sequência
20.
Lipids Health Dis ; 18(1): 86, 2019 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-30947713

RESUMO

BACKGROUND: Cis- and trans-palmitoleic acids (Cis-POA and trans-POA) are isomers of palmitoleic acid, a monounsaturated fatty acid which affects glucose and lipid metabolism, and reduces insulin resistance. Trans-POA is used as a biomarker for indicating the risk of type II diabetes and coronary heart disease, but no methods of analysis or distinguishing between cis-POA and trans-POA have yet been reported. METHOD: An accurate and precise HPLC method was developed to determine cis- and trans-POA simultaneously, and compared with results from a GC method. Cis- and trans-POA were analyzed by HPLC on a reverse-phase BDS-C18 column, equilibrated and eluted with acetonitrile (A) and water (B). In the established and validated GC method used for comparison, potassium hydroxide ester exchange was chosen to derivatize the cis- and trans-POA, before being determined. RESULTS: The calibration curves for cis- and trans-POA were linear over the range 0.05 to 500 µg/mL. The HPLC method exhibited good sensitivity, precision and accuracy. The limits of detection (LOD) for cis- and trans-POA were 0.2 and 0.05 µg/mL, respectively. The method successfully determined cis- and trans-POA in fish oil. For the GC method, the contents of cis-POA quantified were similar to those from the HPLC method, but the contents of trans-POA revealed significant variation between the two methods. CONCLUSIONS: After a comprehensive consideration of the characteristics of the saponification and methyl esterification methods which have been tested and verified, the HPLC method was found to be suitable for determining cis- and trans-POA contents in fish oil. It was also suggested that in natural fish oil, cis-POA may be in the glyceride state, and trans-POA almost completely in the free acid form. In comparison with the GC method, the HPLC method provided a simpler process and faster analyses for identifying and determining cis- and trans-POA. The study has also provided technical support for studying the pharmacological differences and relationship between structure and activity of cis- and trans-POA. This could help physicians to analyze patients' samples more quickly in 10 min and therefore provide a more rapid diagnosis of problems relating to the risk of type II diabetes and coronary heart disease.


Assuntos
Diabetes Mellitus Tipo 2/diagnóstico , Ácidos Graxos Monoinsaturados/isolamento & purificação , Óleos de Peixe/química , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patologia , Ácidos Graxos Monoinsaturados/química , Glucose/metabolismo , Glicerídeos/química , Humanos , Isomerismo , Relação Estrutura-Atividade , Água/química
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