RESUMO
Pien Tze Huang (PZH), a class I nationally protected traditional Chinese medicine (TCM), has been used to treat liver diseases such as hepatitis; however, the effect of PZH on the progression of sepsis is unknown. Here, we reported that PZH attenuated lipopolysaccharide (LPS)-induced sepsis in mice and reduced LPS-induced production of proinflammatory cytokines in macrophages by inhibiting the activation of mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signalling. Mechanistically, PZH stimulated signal transducer and activator of transcription 3 (STAT3) phosphorylation to induce the expression of A20, which could inhibit the activation of NF-κB and MAPK signalling. Knockdown of the bile acid (BA) receptor G protein-coupled bile acid receptor 1 (TGR5) in macrophages abolished the effects of PZH on STAT3 phosphorylation and A20 induction, as well as the LPS-induced inflammatory response, suggesting that BAs in PZH may mediate its anti-inflammatory effects by activating TGR5. Consistently, deprivation of BAs in PZH by cholestyramine resin reduced the effects of PZH on the expression of phosphorylated-STAT3 and A20, the activation of NF-κB and MAPK signalling, and the production of proinflammatory cytokines, whereas the addition of BAs to cholestyramine resin-treated PZH partially restored the inhibitory effects on the production of proinflammatory cytokines. Overall, our study identifies BAs as the effective components in PZH that activate TGR5-STAT3-A20 signalling to ameliorate LPS-induced sepsis.
RESUMO
Fluorescent probes for metal ion recognition can be divided into selective probes, weakly selective probes, and non-selective probes roughly. Weakly selective probes are not often used for quantitative analysis of metal ions due to their overlapping spectra resulting from simultaneous interactions with multiple metal ions. Conversely, the different metal ions contained in herbal medicine extracts from different geographical origins will produce corresponding fluorescence fingerprint profiles after interaction with weakly selective fluorescence probes. The performance can be used in the study of origin tracing of food or Chinese herbal medicine. Weakly selective fluorescent probes of benzimidazole derivatives have been synthesized and attempted to be used in the origin tracing of Radix Astragali in this work. Radix Astragali from different origins will produce different fluorescence fingerprint spectra due to the difference of metal ions and content in combination with the probe. Excitation-emission matrix (EEM) fluorescence spectroscopy in conjunction with N-way partial least squares discriminant analysis (N-PLS-DA), and unfolded partial least squares discriminant analysis (U-PLS-DA) were used to identify the origin of 150 Radix Astragali samples from five geographical origins. The prediction results showed that the correct recognition rates of the U-PLS-DA model and N-PLS-DA model are 95.92% and 93.88%, respectively. In comparison, the results of U-PLS-DA are slightly better than those of N-PLS-DA. These findings indicate that EEM fluorescence spectroscopy based on weakly selective fluorescent probes combined with multi-way chemometrics provides a good idea for the origin tracing of traditional Chinese medicine.
Assuntos
Astragalus propinquus , Medicamentos de Ervas Chinesas , Medicamentos de Ervas Chinesas/química , Corantes Fluorescentes , Quimiometria , Análise dos Mínimos Quadrados , ÍonsRESUMO
Fruit softening, an irreversible process that occurs during fruit ripening, can lead to losses and waste during postharvest transportation and storage. Cell wall disassembly is the main factor leading to loss of fruit firmness, and several ripening-associated cell wall genes have been targeted for genetic modification, particularly pectin modifiers. However, individual knockdown of most cell wall-related genes has had minimal influence on cell wall integrity and fruit firmness, with the notable exception of pectate lyase. Compared to pectin disassembly, studies of the cell wall matrix, the xyloglucan-cellulose framework, and underlying mechanisms during fruit softening are limited. Here, a tomato (Solanum lycopersicum) fruit ripening-associated α-expansin (SlExpansin1/SlExp1) and an endoglucanase (SlCellulase2/SlCel2), which function in the cell wall matrix, were knocked out individually and together using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9-mediated genome editing. Simultaneous knockout of SlExp1 and SlCel2 enhanced fruit firmness, reduced depolymerization of homogalacturonan-type pectin and xyloglucan, and increased cell adhesion. In contrast, single knockouts of either SlExp1 or SlCel2 did not substantially change fruit firmness, while simultaneous overexpression of SlExp1 and SlCel2 promoted early fruit softening. Collectively, our results demonstrate that SlExp1 and SlCel2 synergistically regulate cell wall disassembly and fruit softening in tomato.
Assuntos
Celulase , Solanum lycopersicum , Frutas/metabolismo , Solanum lycopersicum/genética , Celulase/genética , Celulase/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Pectinas/metabolismo , Parede Celular/metabolismoRESUMO
Epulopiscium spp. are the largest known heterotrophic bacteria; a large cigar-shaped individual is a million times the volume of Escherichia coli. To better understand the metabolic potential and relationship of Epulopiscium sp. type B with its host Naso tonganus, we generated a high-quality draft genome from a population of cells taken from a single fish. We propose the name Candidatus Epulopiscium viviparus to describe populations of this best-characterized Epulopiscium species. Metabolic reconstruction reveals more than 5% of the genome codes for carbohydrate active enzymes, which likely degrade recalcitrant host-diet algal polysaccharides into substrates that may be fermented to acetate, the most abundant short-chain fatty acid in the intestinal tract. Moreover, transcriptome analyses and the concentration of sodium ions in the host intestinal tract suggest that the use of a sodium motive force (SMF) to drive ATP synthesis and flagellar rotation is integral to symbiont metabolism and cellular biology. In natural populations, genes encoding both F-type and V-type ATPases and SMF generation via oxaloacetate decarboxylation are among the most highly expressed, suggesting that ATPases synthesize ATP and balance ion concentrations across the cell membrane. High expression of these and other integral membrane proteins may allow for the growth of its extensive intracellular membrane system. Further, complementary metabolism between microbe and host is implied with the potential provision of nitrogen and B vitamins to reinforce this nutritional symbiosis. The few features shared by all bacterial behemoths include extreme polyploidy, polyphosphate synthesis, and thus far, they have all resisted cultivation in the lab.
Assuntos
Sódio , ATPases Vacuolares Próton-Translocadoras , Animais , Sódio/metabolismo , Bactérias/metabolismo , Clostridiales/metabolismo , ATPases Vacuolares Próton-Translocadoras/metabolismo , Trifosfato de Adenosina/metabolismoRESUMO
Several spectroscopic techniques have been used to detect olive oil adulteration. To evaluate the performance of these spectral techniques on this issue, this work performed a comparative study on identifying adulterated olive oil with different concentrations of soybean oil based on Fourier-transform infrared (FTIR), visible-near-infrared (Vis-NIR) and excitation-emission matrix fluorescence spectroscopy (EEMs) combined with chemometrics. Principal component analysis (PCA)/ multi-way-PCA analysis showed the feasibility of the three spectral methods for the identification of olive oil adulteration. The accuracy of FTIR and Vis-NIR based on partial least squares discriminant analysis (PLS-DA) for adulterated olive oil was 100%, while the accuracy of EEMs based on unfold-PLS-DA was only 73%. The accuracy of EEMs combined with back-propagation artificial neural network based on self-weighted alternating trilinear decomposition is 100%. In comparison, FTIR and Vis-NIR are superior for the detection of olive oil adulteration due to the convenience of instrument operation and modeling.
Assuntos
Óleos de Plantas , Óleo de Soja , Azeite de Oliva/análise , Óleo de Soja/análise , Espectrometria de Fluorescência/métodos , Óleos de Plantas/química , Análise de Fourier , Quimiometria , Contaminação de Alimentos/análise , Análise dos Mínimos Quadrados , Espectroscopia de Infravermelho com Transformada de Fourier/métodosRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Herb-derived anti-tumor agents, such as paclitaxel and vincristine, exert significant but varied effectivenesses towards different cancer types. Similarly, Centipeda minima (CM) is a well-known traditional Chinese medicine that has been used to treat rhinitis, relieve pain and reduce swelling, and recently found to exert overwhelming anti-tumor effects against breast cancer, colon cancer, and nasopharyngeal carcinoma with different response rates. However, what is the optimizing cancer model that benefits most from CM, and what is the specific target underlying still require more exclusive and profound investigations. AIMS OF THE STUDY: This study aimed to explore the dominant tumor model and specific target of CM by integrative pharmacology and biological experiments. MATERIALS AND METHODS: The most predominant and specific cancer types that are sensitive to CM were screened and identified based on a combination network pharmacology and bioinformatics analysis. Compound-target network and protein-protein interaction of CM-related cancer targets were carried out to determine the most abundant active compound. Simultaneously, the priority target responsible for CM-related anti-tumor efficacy was further validated by molecular docking and in vitro experiments. RESULTS: In total, approximately 42% (8/19) of the targets were enriched in prostate cancer (p = 1.25E-09), suggesting prostate cancer would be the most sensitive tumor response to CM-related efficacy. Furthermore, we found that arnicolide D (ARD), the most abundant and representative active compound of CM, could directly bind to Src with binding energy of -7.3 kcal/mol, implying Src would be the priority target responsible for CM-related anti-tumor efficacy. Meanwhile, the results were further validated by solvent-induced protein precipitation (SIP) assay. In addition, PCR and WB results also revealed that either CM or ARD could not influence the gene expression of Src, while significantly decreased its protein expression instead, which further suggested that ARD might markedly shortene the Src protein half-life to promote Src protein degradation, thereby achieving significant anti-prostate cancer efficacy. CONCLUSION: Our findings not only suggest CM as a promising Src-targeting candidate for prostate cancer treatment, but also bring up a strategy for understanding the personalization of herbal medicines by using integrative pharmacology.
Assuntos
Asteraceae/química , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes src , Lactonas/farmacologia , Farmacologia em Rede/métodos , Neoplasias da Próstata/tratamento farmacológico , Sesquiterpenos/farmacologia , Transdução de Sinais/efeitos dos fármacos , Antineoplásicos/farmacologia , Disponibilidade Biológica , Bases de Dados Genéticas , Humanos , Masculino , Medicina Tradicional Chinesa , Simulação de Acoplamento Molecular/métodos , Células PC-3 , Compostos Fitoquímicos/farmacologia , Mapas de Interação de ProteínasRESUMO
Prometryn is an occasional triazine herbicide used in aquaculture to kill algae. However, deposition of prometryn at the bottom of the pond poses a potential threat to aquatic animals, especially benthos, such as the sea cucumber. This study investigated the toxic effects of prometryn oral exposure on antioxidants, and the intestinal histomorphology and microbiome of sea cucumbers. Results showed that the accumulation of prometryn in the intestine, respiratory tree, and body wall decreased sequentially under the same level. Severe pathological damages were observed in the intestines of sea cucumbers fed with 0.080 and 1.595 g/kg prometryn (measured concentration). Moreover, hydrogen peroxide (H2O2) and malondialdehyde (MDA) concentrations were significantly increased in prometryn treatment groups compared to the control group (P < 0.05), while the catalase (CAT) activity was significantly decreased (P < 0.05) in the coelomic fluid of treatment groups. At the phylum level, the abundance of Proteobacteria was significantly higher in the 0.080 g/kg treatment group than in the control group. In addition, prometryn exposure reduced the diversity of intestinal microflora in sea cucumbers. In conclusion, these results suggest that prometryn has potential toxicity to sea cucumber. Therefore, the harm of prometryn deposited in the sediment to aquatic animals must be a concern in aquaculture.
Assuntos
Ração Animal/análise , Antioxidantes/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Prometrina/toxicidade , Pepinos-do-Mar/efeitos dos fármacos , Animais , Suplementos Nutricionais , Herbicidas/toxicidade , Imunidade Inata , Intestinos/efeitos dos fármacos , Intestinos/microbiologia , Intestinos/patologia , Pepinos-do-Mar/imunologia , Pepinos-do-Mar/metabolismo , Pepinos-do-Mar/microbiologiaRESUMO
Chinese water chestnut peels are a kind of vegetable processing waste containing many active components such as polysaccharides, the structure of which remains unknown. To elucidate the structure of polysaccharides from Chinese water chestnut peels, two polysaccharides named WVP-1 and WVP-2 were isolated. WVP-1 (3.16 kDa) consisted of mannose (1.75 %), glucose (84.69 %), galactose (6.32 %), and arabinose (7.24 %), while WVP-2 (56.97 kDa) was composed of mannose (3.18 %), rhamnose (1.52 %), glucuronic acid (1.42 %), galacturonic acid (4.83 %), glucose (11.51 %), galactose (36.02 %), and arabinose (41.53 %). Linkage and NMR data indicated that WVP-1 was composed mainly of â4)-α-d-Glcp(1â and a certain proportion of â3)-ß-d-Glcp-(1â, including linear and branched polysaccharides simultaneously. WVP-2 was a pectin-like polysaccharide with â4)-α-d-GalpA6Me-(1â units and the branch points of â3,4)-α-l-Arap-(1â, â3,6)-ß-d-Galp-(1â. WVP-2 exhibited stronger potential antioxidant and immunomodulatory activities than WVP-1 in vitro. These results provide a foundation for the further study of polysaccharides from Chinese water chestnut peels.
Assuntos
Antioxidantes/química , Eleocharis/química , Fatores Imunológicos/química , Extratos Vegetais/química , Polissacarídeos/química , Animais , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Ácido Glucurônico , Ácidos Hexurônicos , Fatores Imunológicos/isolamento & purificação , Fatores Imunológicos/farmacologia , Interleucina-6/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Camundongos , Peso Molecular , Monossacarídeos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Células RAW 264.7 , Ramnose , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Cryo-electron tomography combined with subtomogram averaging (StA) has yielded high-resolution structures of macromolecules in their native context. However, high-resolution StA is not commonplace due to beam-induced sample drift, images with poor signal-to-noise ratios (SNR), challenges in CTF correction, and limited particle number. Here we address these issues by collecting tilt series with a higher electron dose at the zero-degree tilt. Particles of interest are then located within reconstructed tomograms, processed by conventional StA, and then re-extracted from the high-dose images in 2D. Single particle analysis tools are then applied to refine the 2D particle alignment and generate a reconstruction. Use of our hybrid StA (hStA) workflow improved the resolution for tobacco mosaic virus from 7.2 to 4.4 Å and for the ion channel RyR1 in crowded native membranes from 12.9 to 9.1 Å. These resolution gains make hStA a promising approach for other StA projects aimed at achieving subnanometer resolution.
Assuntos
Canais de Cálcio/química , Microscopia Crioeletrônica/métodos , Processamento de Imagem Assistida por Computador/métodos , Tomografia com Microscopia Eletrônica/métodos , Substâncias Macromoleculares/química , Canal de Liberação de Cálcio do Receptor de Rianodina/química , Razão Sinal-Ruído , Imagem Individual de Molécula , Fluxo de TrabalhoRESUMO
Curcumin is the main secondary metabolite of Curcuma longa and other Curcuma spp, and has been reported to have some potential in preventing and treating some physiological disorders. This study investigated the effect of curcumin in inhibiting high-fat diet and streptozotocin (STZ)-induced hyperglycemia and hyperlipidemia in rats. Twenty-six male Sprague-Dawley (SD) rats (170-190 g) were randomly divided into a standard food pellet diet group (Control group), a high-fat diet and streptozotocin group (HF + STZ group), and a high-fat diet combined with curcumin and STZ group (HF + Cur + STZ group). Compared with the HF + STZ group, the HF + Cur + STZ group exhibited significantly reduced fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), alanine aminotransferase (AST), and aspartate transaminase (ALT) levels, as well as liver coefficients. In the livers of these rats, the expression of malondialdehyde (MDA) and Bax was downregulated, whereas that of superoxide dismutase (SOD) and Bcl-2 was upregulated. Moreover, the liver histology of these rats was improved and resembled that of the control rats. These results suggest that curcumin prevents high-fat diet and STZ-induced hyperglycemia and hyperlipidemia, mainly via anti-oxidant and anti-apoptotic mechanisms in the liver.
Assuntos
Curcumina/farmacologia , Hiperglicemia/tratamento farmacológico , Hiperlipidemias/tratamento farmacológico , Extratos Vegetais/farmacologia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Glicemia , Curcuma/química , Dieta Hiperlipídica/efeitos adversos , Humanos , Hiperglicemia/sangue , Hiperglicemia/induzido quimicamente , Hiperglicemia/patologia , Hiperlipidemias/sangue , Hiperlipidemias/induzido quimicamente , Hiperlipidemias/patologia , Lipídeos/sangue , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Extratos Vegetais/química , Ratos , Estreptozocina/toxicidade , Superóxido Dismutase/metabolismo , Triglicerídeos/sangueRESUMO
A novel polysaccharide designated SLP-4 with the Mw of 19681 Da was purified from the petal of Saussurea laniceps. Monosaccharide composition analysis indicated that SLP-4 was composed of mannose, rhamnose, galacturonic acid, glucose, galactose, xylose and arabinose in a molar ratio of 0.825:2.030:14.998:0.841:8.260:4.039:6.009. Structural features indicated that SLP-4 was a typical pectin polysaccharide with a backbone containing â3,6)-Galp-(1â, â4)-GalpA-(1â, â6)-Galp-(1â, â4, 6)-Galp-(1â and â2, 4)-Rhap-(1â with the branches of â4)-Galp-(1â, T-Galp-(1â, â3)-Galp-(1â, T-Rhap-(1â, T-Araf-(1â, â5)-Araf-(1â, T-Glcp-(1â, â4)-Xylp-(1â and â4)-Manp-(1â. Additionally, SLP-4 could effectively inhibit the secretion of HBsAg and HBeAg in HepG2.2.15 cells, but had little effect on the replication of HBV DNA. This inhibition didn't involve cellular pathways, and was due to the interaction between SLP-4 and HBsAg or HBeAg, which may block the ELISA detection of HBsAg and HBeAg. The present study may provide useful information for further study of SLP-4 and understanding of anti-HBV activity of polysaccharides.
Assuntos
Antivirais/farmacologia , Frutas/química , Vírus da Hepatite B/efeitos dos fármacos , Pectinas/farmacologia , Polissacarídeos/farmacologia , Saussurea/química , Antivirais/química , Antivirais/isolamento & purificação , Configuração de Carboidratos , Testes de Sensibilidade Microbiana , Tamanho da Partícula , Pectinas/química , Pectinas/isolamento & purificação , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Propriedades de SuperfícieRESUMO
BACKGROUND: Moderate electric field (MEF) technology is a promising food preservation strategy since it relies on physical properties-rather than chemical additives-to preserve solid cellular foods during storage. However, the effectiveness of long-term MEF exposure on the psychrotrophic microorganisms responsible for the food spoilage at cool temperatures remains unclear. RESULTS: The spoilage-associated psychrotroph Pseudomonas fragi MC16 was obtained from pork samples stored at 7 °C. Continuous MEF treatment attenuated growth and resulted in subsequent adaptation of M16 cultured on nutrient agar plates at 7 °C, compared to the control cultures, as determined by biomass analysis and plating procedures. Moreover, intracellular dehydrogenase activity and ATP levels also indicated an initial effect of MEF treatment followed by cellular recovery, and extracellular ß-galactosidase activity assays indicated no obvious changes in cell membrane permeability. Furthermore, microscopic observations using scanning and transmission electron microscopy revealed that MEF induced sublethal cellular injury during early treatment stages, but no notable changes in morphology or cytology on subsequent days. CONCLUSION: Our study provides direct evidence that psychrotrophic P. fragi MC16 cultured on nutrient agar plates at 7 °C are capable of adapting to MEF treatment.
Assuntos
Eletricidade , Microbiologia de Alimentos , Conservação de Alimentos/métodos , Pseudomonas fragi/crescimento & desenvolvimento , Pseudomonas fragi/metabolismo , Pseudomonas fragi/efeitos da radiação , Trifosfato de Adenosina/análise , Animais , Biomassa , Permeabilidade da Membrana Celular/fisiologia , Permeabilidade da Membrana Celular/efeitos da radiação , Contagem de Colônia Microbiana , Terapia por Estimulação Elétrica , Ativação Enzimática , Ensaios Enzimáticos , Armazenamento de Alimentos , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Oxirredutases/metabolismo , Oxirredutases/efeitos da radiação , Pseudomonas fragi/enzimologia , Carne Vermelha/microbiologia , Refrigeração , Suínos , Temperatura , beta-Galactosidase/metabolismo , beta-Galactosidase/efeitos da radiaçãoRESUMO
In this study, a selenoprotein W cDNA was cloned from topmouth culter (Erythroculter ilishaeformis), and it was designated as EISelW. The EISelW open reading frame was composed of 261 base pairs (bp), encoding 86-amino-acid protein. The 5' untranslated region (UTR) consisted of 104 bp, and the 3'-UTR was composed of 365 bp. A selenocysteine insertion sequence (SECIS) element was found in the 3'-UTR of EISelW mRNA. The SECIS element was classified as form II because of a small additional apical loop presented in SECIS element of EISelW mRNA. Bioinformatic approaches showed that the secondary structure of EISelW was a ß1-α1-ß2-ß3-ß4-α2 pattern from amino-terminal to carboxy-terminal. Real-time PCR analysis of EISelW mRNAs expression in 17 tissues showed that the EISelW mRNA was predominantly expressed in liver, ovary, pituitary, various regions of the brain, spinal cord and head kidney. Study of intraperitoneal injection showed that the levels of EISelW mRNA in brain, liver, ovary and spleen were regulated by somatostatin 14 (SS14), 17ß-estradiol (E2), cysteamine hydrochloride (CSH) and a binary mixture of E2 and CSH, dependent on the dosage. These results suggest that E2, SS14 and CSH status may affect tissues of selenium metabolism by regulating the expression of SelW mRNA, as SelW plays a central role in selenium metabolism.
Assuntos
Cisteamina/farmacologia , Estradiol/farmacologia , Perciformes/genética , Selenoproteína W/genética , Somatostatina/farmacologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Encéfalo/metabolismo , DNA Complementar/genética , Interações Medicamentosas , Feminino , Fígado/metabolismo , Masculino , Ovário/metabolismo , Filogenia , RNA Mensageiro/metabolismo , Baço/metabolismoRESUMO
Acute lung injury (ALI) is a major cause of acute respiratory failure in critically-ill patients. Resveratrol and curcumin are proven to have potent anti-inflammatory efficacy, but their clinical application is limited by their metabolic instability. Here, a series of resveratrol and the Mono-carbonyl analogs of curcumin (MCAs) hybrids were designed and synthesized by efficient aldol construction strategy, and then screened for anti-inflammatory activities in vitro and in vivo. The results showed that the majority of analogs effectively inhibited the LPS-induced production of IL-6 and TNF-α. Five analogs, a9, a18, a19, a20 and a24 exhibited excellent anti-inflammatory activity in a dose-dependent manner along with low toxicity in vitro. Structure activity relationship study revealed that the electron-withdrawing groups at meta-position and methoxyl group (OCH3) at the para position of the phenyl ring were important for anti-inflammatory activities. The most promising compound a18 decreased LPS induced TNF-α, IL-6, IL-12, and IL-33 mRNA expression. Additionally, a18 significantly protected against LPS-induced acute lung injury in the in vivo mouse model. The research of resveratrol and MCAs hybrids could bring insight into the treatment of inflammatory diseases and compound a18 may serve as a lead compound for the development of anti-ALI agents.
Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Anti-Inflamatórios/química , Anti-Inflamatórios/uso terapêutico , Curcumina/análogos & derivados , Curcumina/uso terapêutico , Estilbenos/química , Estilbenos/uso terapêutico , Lesão Pulmonar Aguda/genética , Lesão Pulmonar Aguda/imunologia , Lesão Pulmonar Aguda/patologia , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-6/genética , Interleucina-6/imunologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pulmão/patologia , Camundongos , Células RAW 264.7 , RNA Mensageiro/genética , Resveratrol , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Elephantopus mollis (EM) is a traditional herbal medicine with multiple pharmacological activities. However, the efficacy of EM in treating human leukemia is currently unknown. In the current study, we report that EM23, a natural sesquiterpene lactone isolated from EM, inhibits the proliferation of human chronic myeloid leukemia (CML) K562 cells and acute myeloid leukemia (AML) HL-60 cells by inducing apoptosis. Translocation of membrane-associated phospholipid phosphatidylserines, changes in cell morphology, activation of caspases, and cleavage of PARP were concomitant with this inhibition. The involvement of the mitochondrial pathway in EM23-mediated apoptosis was suggested by observed disruptions in mitochondrial membrane potential. Mechanistic studies indicated that EM23 caused a marked increase in the level of reactive oxygen species (ROS). Pretreatment with N-acetyl-L-cysteine, a ROS scavenger, almost fully reversed EM23-mediated apoptosis. In EM23-treated cells, the expression levels of thioredoxin (Trx) and thioredoxinreductase (TrxR), two components of the Trx system involved in maintaining cellular redox homeostasis, were significantly down-regulated. Concomitantly, Trx regulated the activation of apoptosis signal-regulating kinase 1 (ASK1) and its downstream regulatory targets, the p38, JNK, and ERK MAPKs. EM23-mediated activation of ASK1/MAPKs was significantly inhibited in the presence of NAC. Furthermore, tumor necrosis factor alpha (TNF-α)-mediated activation of nuclear factor-κB (NF-κB) was suppressed by EM23, as suggested by the observed blockage of p65 nuclear translocation, phosphorylation, and reversion of IκBα degradation following EM23 treatment. Taken together, these results provide important insights into the anticancer activities of the EM component EM23 against human CML K562 cells and AML HL-60 cells.
RESUMO
Sesquiterpene lactones (SLs) are the active constituents of a variety of medicinal plants and found to have potential anticancer activities. However, the intracellular molecular targets of SLs and the underlying molecular mechanisms have not been well elucidated. In this study, we observed that EM23, a natural SL, exhibited anti-cancer activity in human cervical cancer cell lines by inducing apoptosis as indicated by caspase 3 activation, XIAP downregulation and mitochondrial dysfunction. Mechanistic studies indicated that EM23-induced apoptosis was mediated by reactive oxygen species (ROS) and the knockdown of thioredoxin (Trx) or thioredoxin reductase (TrxR) resulted in a reduction in apoptosis. EM23 attenuated TrxR activity by alkylation of C-terminal redox-active site Sec498 of TrxR and inhibited the expression levels of Trx/TrxR to facilitate ROS accumulation. Furthermore, inhibition of Trx/TrxR system resulted in the dissociation of ASK1 from Trx and the downstream activation of JNK. Pretreatment with ASK1/JNK inhibitors partially rescued cells from EM23-induced apoptosis. Additionally, EM23 inhibited Akt/mTOR pathway and induced autophagy, which was observed to be proapoptotic and mediated by ROS. Together, these results reveal a potential molecular mechanism for the apoptotic induction observed with SL compound EM23, and emphasize its putative role as a therapeutic agent for human cervical cancer.
Assuntos
Lactonas/farmacologia , MAP Quinase Quinase 4/metabolismo , Sesquiterpenos/farmacologia , Tiorredoxina Dissulfeto Redutase/antagonistas & inibidores , Tiorredoxina Dissulfeto Redutase/metabolismo , Neoplasias do Colo do Útero/tratamento farmacológico , Apoptose/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Ativação Enzimática , Feminino , Células HL-60 , Humanos , Células K562 , Células MCF-7 , Espécies Reativas de Oxigênio/metabolismo , Transfecção , Neoplasias do Colo do Útero/enzimologia , Neoplasias do Colo do Útero/patologiaRESUMO
In this study, the cDNA encoding insulin-like growth factor-binding protein-1 (IGFBP-1) was cloned from the liver of goldfish (Carassius auratus). The obtained goldfish IGFBP-1 cDNA sequence was 1037 bp in length and had an open reading frame of 789 bp encoding a predicted polypeptide of 262 amino acid residues. IGFBP-1 transcript was detected in all tested central nervous and peripheral tissues. The relatively higher levels of IGFBP-1 mRNA were observed in the liver, gill, kidney, heart, spleen, fat and testis, while the lower levels were found in all different regions of brain, muscle and intestine. In the skin, IGFBP-1 mRNA expression level was extremely low. The IGFBP-1 mRNA expression level in liver was significantly elevated after feeding. With cadmium exposure for 24 h, IGFBP-1 mRNA expression levels in spleen and liver were significantly increased at different cadmium concentrations ranging from 0.5 to 10 ppm. The results in this study provided the data regarding molecular characteristics and expression patterns of IGFBP-1 in goldfish and showed that the expression of IGFBP-1 mRNA might be associated with metabolic status and heavy metal stress and regulated by metabolic factors and cadmium in fish.
Assuntos
Cádmio/toxicidade , Proteínas de Peixes/genética , Carpa Dourada/genética , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Poluentes Químicos da Água/toxicidade , Animais , Clonagem Molecular , DNA Complementar/genética , Ingestão de Alimentos/genética , RNA Mensageiro/metabolismoRESUMO
Acute lung injury (ALI) is a leading cause of morbidity and mortality in critically-ill patients. Previously, we reported that a symmetric mono-carbonyl analog of curcumin, (C66), exhibits enhanced stability and was found to have efficacy and be involved in potential cytokines inhibition. In the present study, a series of novel asymmetric C66 analogs were designed and synthesized. A majority of them effectively inhibited the LPS-induced expression of TNF-α and IL-6. Significantly, compound 4b2 was found to effectively reduce LPS-induced pulmonary inflammation, as reflected by reductions in concentration of total protein, inflammatory cell count as well as the lung W/D ratio in bronchoalveolar lavage (BAL) fluid. Furthermore, in vivo administration of 4b2 resulted in remarkable improvement in histopathological changes of lung in rats.
Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Anti-Inflamatórios não Esteroides/farmacologia , Curcumina/farmacologia , Desenho de Fármacos , Anti-Inflamatórios não Esteroides/síntese química , Anti-Inflamatórios não Esteroides/química , Curcumina/síntese química , Curcumina/química , Citocinas/antagonistas & inibidores , Citocinas/biossíntese , Relação Dose-Resposta a Droga , Humanos , Lipopolissacarídeos/antagonistas & inibidores , Lipopolissacarídeos/farmacologia , Estrutura Molecular , Relação Estrutura-AtividadeRESUMO
We previously reported a symmetric monocarbonyl analog of curcumin (MACs), C66, which demonstrated potential anti-inflammatory activity and low toxicity. In continuation of our ongoing research, we designed and synthesized 34 asymmetric MACs based on C66 as a lead molecule. A majority of the C66 analogs effectively inhibited LPS induction of TNF-α and IL-6 expression. Additionally, a preliminary SAR was conducted. Furthermore, active compounds 4a11 and 4a16 were found to effectively reduce the W/D ratio in the lungs and the protein concentration in the bronchoalveolar lavage fluid (BALF). Meanwhile, a histopathological examination indicated that these two analogs significantly attenuate tissue injury in the lungs with LPS-induced ALI rats. 4a11 and 4a16 also inhibited mRNA expression of several inflammatory cytokines, including TNF-α, IL-6, IL-1ß, COX-2, ICAM-1 and VCAM-1, in the Beas-2B cells after LPS challenge. Altogether, the data exhibit a series of new C66 analogs as promising anti-inflammatory agents for the treatment of LPS-induced ALI.
Assuntos
Lesão Pulmonar Aguda/tratamento farmacológico , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/farmacologia , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/patologia , Animais , Anti-Inflamatórios não Esteroides/síntese química , Compostos de Benzilideno/química , Líquido da Lavagem Broncoalveolar , Técnicas de Química Sintética , Cicloexanonas/química , Citocinas/genética , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Estabilidade de Medicamentos , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolissacarídeos/toxicidade , Masculino , Ratos Sprague-Dawley , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/metabolismoRESUMO
In the present study, a full-length cDNA encoding the insulin-like growth factor binding protein-2 (IGFBP-2) was cloned from the liver of goldfish (Carassius auratus) by rapid amplification of cDNA ends technique. The goldfish IGFBP-2 cDNA sequence was 1,513 bp long and had an open reading frame of 825 bp encoding a predicted polypeptide of 274 amino acid residues. Semi-quantitative RT-PCR results revealed that goldfish IGFBP-2 mRNA was expressed in all detected tissues. In liver, central nervous system and pituitary gland, goldfish IGFBP-2 expressed at high levels, followed by anterior intestine, middle intestine and kidney. In posterior intestine, ovary, skin, fat, spleen, muscle and gill, the goldfish IGFBP-2 expression levels were very low. Fasting and refeeding experiment showed that the mRNA expression of goldfish IGFBP-2 was up-regulated significantly in liver compared to the fed group and restored rapidly to normal level after refed. However, the mRNA expressions of IGFBP-2 in hypothalamus and pituitary of goldfish were insensitive to fasting. Furthermore, the mRNA expressions of IGFBP-2 in hypothalamus, pituitary and liver were varied in periprandial changes and significantly down-regulated at 2 and 4 h after meal. These results imply that the IGFBP-2 mRNA expression may be associated with anabolic and catabolic metabolism and regulated by metabolic factors in goldfish.