RESUMO
Developing more effective bioactive ingredients of natural origin is imperative for promoting wound healing. Sea cucumbers have long enjoyed a good reputation as both food delicacies and traditional medicines. In this study, we heterogeneously expressed a Apostichopus japonicus derived novel protein AjPSPLP-3, which exhibits a theoretical molecular weight of 13.034 kDa, through fusion with maltose binding protein (MBP). AjPSPLP-3 contains a strict CXXCXC motif, nine extremely conserved cysteine residues and two highly conserved cysteine residues. The predicted structure of AjPSPLP-3 consists of random coil and nine ß-sheets, Cys30-Cys67, Cys38-Cys58, Cys53-Cys90, Cys56-Cys66, and Cys81-Cys102 participating in the formation of five pairs of disulfide bonds. In vitro experiments conducted on HaCaT cells proved that AjPSPLP-3 and MBP-fused AjPSPLP-3 significantly contribute to HaCaT cells proliferation and migration without exhibiting hemolytic activity on murine erythrocytes. Specifically, treatment with 10 µmol/L MBP-fused AjPSPLP-3 protein increased the viability of HaCaT cells by 12.28 % (p < 0.001), while treatment with 10 µmol/L AjPSPLP-3 protein increased viability of HaCaT cells by 6.01 % (p < 0.01). Furthermore, wound closure of MBP-fused AjPSPLP-3 and AjPSPLP-3 were 22.51 % (p < 0.01) and 7.32 % (p < 0.05) higher than that of the control groups in HaCaT cells following 24 h of incubation.
Assuntos
Movimento Celular , Proliferação de Células , Stichopus , Animais , Stichopus/genética , Stichopus/química , Proliferação de Células/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Humanos , Camundongos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/farmacologia , Proteínas Recombinantes de Fusão/metabolismo , Proteínas Recombinantes de Fusão/isolamento & purificação , Clonagem Molecular , Sequência de Aminoácidos , Linhagem Celular , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Ligantes de Maltose/genética , Proteínas Ligantes de Maltose/química , Proteínas Ligantes de Maltose/metabolismo , Células HaCaTRESUMO
Sea cucumber viscera contain various naturally occurring active substances, but they are often underutilized during sea cucumber processing. Polydeoxyribonucleotide (PDRN) is an adenosine A2A receptor agonist that activates the A2A receptor to produce various biological effects. Currently, most studies on the activity of PDRN have focused on its anti-inflammatory, anti-apoptotic, and tissue repair properties, yet relatively few studies have investigated its antioxidant activity. In this study, we reported for the first time that PDRN was extracted from the sperm of Apostichopus japonicus (AJS-PDRN), and we evaluated its antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), and hydroxyl radical scavenging assays. An in vitro injury model was established using H2O2-induced oxidative damage in RAW264.7 cells, and we investigated the protective effect of AJS-PDRN on these cells. Additionally, we explored the potential mechanism by which AJS-PDRN protects RAW264.7 cells from damage using iTRAQ proteomics analysis. The results showed that AJS-PDRN possessed excellent antioxidant activity and could significantly scavenge DPPH, ABTS, and hydroxyl radicals. In vitro antioxidant assays demonstrated that AJS-PDRN was cytoprotective and significantly enhanced the antioxidant capacity of RAW264.7 cells. The results of GO enrichment and KEGG pathway analysis indicate that the protective effects of AJS-PDRN pretreatment on RAW264.7 cells are primarily achieved through the regulation of immune and inflammatory responses, modulation of the extracellular matrix and signal transduction pathways, promotion of membrane repair, and enhancement of cellular antioxidant capacity. The results of a protein-protein interaction (PPI) network analysis indicate that AJS-PDRN reduces cellular oxidative damage by upregulating the expression of intracellular selenoprotein family members. In summary, our findings reveal that AJS-PDRN mitigates H2O2-induced oxidative damage through multiple pathways, underscoring its significant potential in the prevention and treatment of diseases caused by oxidative stress.
Assuntos
Antioxidantes , Peróxido de Hidrogênio , Estresse Oxidativo , Polidesoxirribonucleotídeos , Proteômica , Espermatozoides , Animais , Camundongos , Peróxido de Hidrogênio/toxicidade , Proteômica/métodos , Masculino , Antioxidantes/farmacologia , Antioxidantes/isolamento & purificação , Estresse Oxidativo/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/metabolismo , Células RAW 264.7 , Polidesoxirribonucleotídeos/farmacologia , Stichopus/química , Pepinos-do-Mar/química , Substâncias Protetoras/farmacologiaRESUMO
Three polysaccharides (SnNG, SnFS and SnFG) were purified from the body wall of Stichopus naso. The physicochemical properties, including monosaccharide composition, molecular weight, sulfate content, and optical rotation, were analyzed, confirming that SnFS and SnFG are sulfated polysaccharides commonly found in sea cucumbers. The highly regular structure {3)-L-Fuc2S-(α1,}n of SnFS was determined via a detailed NMR analysis of its oxidative degradation product. By employing ß-elimination depolymerization of SnFG, tri-, penta-, octa-, hendeca-, tetradeca-, and heptadeca-saccharides were obtained from the low-molecular-weight product. Their well-defined structures confirmed that SnFG possessed the backbone of {D-GalNAc4S6S-ß(1,4)-D-GlcA}, and each GlcA residue was branched with Fuc2S4S. SnFS and SnFG are both structurally the simplest version of natural fucan sulfate and fucosylated glycosaminoglycan, facilitating the application of low-value sea cucumbers S. naso. Bioactivity assays showed that SnFG and its derived oligosaccharides exhibited potent anticoagulation and intrinsic factor Xase (iXase) inhibition. Moreover, a comparative analysis with the series of oligosaccharides solely branched with Fuc3S4S showed that in oligosaccharides with lower degrees of polymerization, such as octasaccharides, Fuc2S4S led to a greater increase in APTT prolongation and iXase inhibition. As the degree of polymerization increases, the influence from the sulfation pattern diminishes, until it is overshadowed by the effects of molecular weight.
Assuntos
Anticoagulantes , Peso Molecular , Oligossacarídeos , Polissacarídeos , Animais , Anticoagulantes/farmacologia , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Polissacarídeos/farmacologia , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Oligossacarídeos/farmacologia , Oligossacarídeos/química , Oligossacarídeos/isolamento & purificação , Stichopus/química , Pepinos-do-Mar/química , Sulfatos/química , Espectroscopia de Ressonância Magnética , Coagulação Sanguínea/efeitos dos fármacosRESUMO
Long-term and excessive alcohol consumption can lead to the development of alcoholic liver disease (ALD), characterized by oxidative damage, intestinal barrier injury, and disruption of intestinal microbiota. In this study, we extracted fucoidan (Aj-FUC) from Apostichopus japonicus using enzymatic methods and characterized its structure. The ALD model was established in male Balb/c mice using 56° Baijiu, with silymarin as a positive control. Mice were orally administered 100 mg/kg·bw and 300 mg/kg·bw of Aj-FUC for 28 days to evaluate its effects on liver injury in ALD mice and explore its potential role in modulating the gut-liver axis. The results showed significant improvements in histopathological changes and liver disease in the Aj-FUC group. Aj-FUC treatment significantly increased the levels of glutathione (GSH) and glutathione peroxidase (GSH-Px) while weakly reduced the elevation of malondialdehyde (MDA) induced by ALD. It also regulated the Nrf2/HO-1 signaling pathway, collectively alleviating hepatic oxidative stress. Aj-FUC intervention upregulated the expression of ZO-1 and Occludin, thus contributing to repair the intestinal barrier. Additionally, Aj-FUC increased the content of short-chain fatty acids (SCFAs) and regulated the imbalance in gut microbiota. These results suggested that Aj-FUC alleviates ALD by modulating the gut-liver axis homeostasis. It may prove to be a useful dietary supplement in the treatment of alcoholic liver damage.
Assuntos
Homeostase , Hepatopatias Alcoólicas , Fígado , Estresse Oxidativo , Polissacarídeos , Stichopus , Animais , Hepatopatias Alcoólicas/tratamento farmacológico , Hepatopatias Alcoólicas/metabolismo , Hepatopatias Alcoólicas/patologia , Polissacarídeos/farmacologia , Polissacarídeos/química , Camundongos , Masculino , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Homeostase/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Stichopus/química , Camundongos Endogâmicos BALB C , Malondialdeído/metabolismo , Microbioma Gastrointestinal/efeitos dos fármacos , Glutationa/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Transdução de Sinais/efeitos dos fármacos , Modelos Animais de Doenças , Glutationa Peroxidase/metabolismoRESUMO
Apostichopus japonicus (A. japonicus) has rich nutritional value and is an important economic crop. Due to its rich endogenous enzyme system, fresh A. japonicus is prone to autolysis during market circulation and storage, resulting in economic losses. In order to alleviate this phenomenon, we investigated the effect of polyphenol oxidase (PPO) mediated (-)-epigallocatechin gallate (EGCG) on the activity and structure of endogenous cathepsin series protein (CEP) from A. japonicus. Research on cathepsin activity showed that PPO mediated EGCG could significantly reduce enzyme activity, resulting in a decrease in enzymatic reaction rate. SDS-PAGE and scanning electron microscopy results showed that PPO mediates EGCG could induce CEP aggregation to form protein aggregates. Various spectral results indicated that EGCG caused changes in the structure of CEP. Meanwhile, the conjugates formed by PPO mediated EGCG had lower thermal stability. In conclusion, PPO mediated EGCG was an effective method to inhibit the endogenous enzyme activity.
Assuntos
Catequina , Catequina/análogos & derivados , Catecol Oxidase , Catepsinas , Stichopus , Catequina/química , Catequina/farmacologia , Catecol Oxidase/metabolismo , Catecol Oxidase/química , Animais , Stichopus/enzimologia , Stichopus/química , Catepsinas/metabolismo , Catepsinas/química , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/química , Estabilidade Enzimática , CinéticaRESUMO
Sea cucumbers release chemical repellents from their guts when they are in danger from predators or a hostile environment. To investigate the chemical structure of the repellent, we collected and chemically analyzed the viscera of stressed sea cucumbers (Apostichopus japonicus) in the Yellow Sea of China. Two undescribed triterpene glycosides (1 and 2), together with a known cladoloside A (3), were identified and elucidated as 3ß-O-{2-O-[ß-d-quinovopyranosyl]-4-O-[3-O-methyl-ß-d-glucopyranosyl-(1â3)-ß-d-glucopyranosyl]-ß-d-xylopyranosyl}-holosta-9(11),25(26)-dien-16-one (1), 3ß-O-{2-O-[ß-d-glucopyranosyl]-4-O-[3-O-methyl-ß-d-glucopyranosyl-(1â3)-ß-d-glucopyranosyl]-ß-d-xylopyranosyl}-holosta-9(11),25(26)-dien-16-one (2), 3ß-O-{2-O-[3-O-methyl-ß-d-glucopyranosyl-(1â3)-ß-d-xylopyranosyl-(1â4)-ß-d-quinovopyranosyl]-ß-d-xylopyranosyl}-holosta-9(11),25(26)-dien-16-one (3) by spectroscopic analysis, including HR-ESI-MS and NMR spectra. Compounds 1, 2, and 3 display embryonic toxicity, as indicated by their 96-hour post-fertilization lethal concentration (96 hpf-LC50) values of 0.289, 0.536, and 0.091â µM, respectively. Our study discovered a class of triterpene glycoside compounds consisting of an oligosaccharide with four sugar units and a holostane aglycone. These compounds possess embryotoxicity and may serve as chemical defense molecules in marine benthic ecosystems.
Assuntos
Glicosídeos , Triterpenos , Animais , Glicosídeos/química , Glicosídeos/isolamento & purificação , Glicosídeos/toxicidade , Triterpenos/química , Triterpenos/isolamento & purificação , Triterpenos/farmacologia , Stichopus/química , Vísceras/química , Pepinos-do-Mar/química , Embrião não Mamífero/efeitos dos fármacosRESUMO
Fibrillin is an important structural protein in connective tissues. The presence of fibrillin in sea cucumber Apostichopus japonicus is still poorly understood, which limits our understanding of the role of fibrillin in the A. japonicus microstructure. The aim of this study was to clarify the presence of fibrillin in the sea cucumber A. japonicus body wall. Herein, the presence of fibrillin in sea cucumber A. japonicus was investigated by utilizing targeted proteomics and visualization strategies. The contents of three different isoforms of fibrillin with high abundance in A. japonicus were determined to be 0.96, 2.54, and 0.15 µg/g (wet base), respectively. The amino acid sequence of fibrillin (GeneBank number: PIK56741.1) that started at position 631 and ended at position 921 was selected for cloning and expressing antigen. An anti-A. japonicus fibrillin antibody with a titer greater than 1:64â¯000 was successfully obtained. It was observed that the distribution of fibrillin in the A. japonicus body wall was scattered and dispersed in the form of fibril bundles at the microscale. It further observed that fibrillin was present near collagen fibrils and some entangled outside the collagen fibrils at the nanoscale. Moreover, the stoichiometry of the most dominant collagen and fibrillin molecules in A. japonicus was determined to be approximately 250:1. These results contribute to an understanding of the role of fibrillin in the sea cucumber microstructure.
Assuntos
Pepinos-do-Mar , Stichopus , Animais , Stichopus/genética , Stichopus/química , Pepinos-do-Mar/metabolismo , Proteômica , Fibrilinas , Colágeno/químicaRESUMO
This study aimed to investigate the alleviative effects of fucosylated chondroitin sulfate from sea cucumber Stichopus chloronotus (fCSSc) on the intestinal barrier injury and oxidative stress damage in vitro and in vivo. The results showed that fCS-Sc protected the intestinal barrier and improved the antioxidant function in H2O2 damaged Caco-2 cells via up-regulating the tight junction proteins and activating Keap1-Nrf2-ARE antioxidant pathway. Furthermore, administration fCS-Sc could ameliorate the weight loss and spleen index decrease in Cyclophosphamide (Cy) treated mice, improve the expressions of ZO-1, Claudin-1, Nrf2, SOD, and NQO-1 in Cy damaged colon tissue, showing significant protective effects against intestinal barrier damage and oxidative stress in vivo. fCS-Sc intervention also alleviated the gut microbiota disorder though increasing the richness and diversity of intestinal bacteria, regulating the structural composition of gut microbiota. fCS-Sc promoted the relative abundance of beneficial microbiota and inhibited the growth of harmful bacteria. This study provided a theoretical basis for the application of fCS-Sc as a prebiotic in chemotherapy.
Assuntos
Pepinos-do-Mar , Stichopus , Humanos , Animais , Camundongos , Stichopus/química , Pepinos-do-Mar/química , Proteína 1 Associada a ECH Semelhante a Kelch , Antioxidantes , Células CACO-2 , Peróxido de Hidrogênio , Fator 2 Relacionado a NF-E2 , Sulfatos de Condroitina/farmacologia , Sulfatos de Condroitina/química , Estresse OxidativoRESUMO
Apostichopus japonicus, also known as Stichopus japonicus, with medicinal and food homologous figures, is a globally recognized precious ingredient with extremely high nutritional value. There is no relevant review available through literature search, so this article selects the research articles through the keywords "sea cucumber" and "Apostichopus japonicus (Stichopus japonicus)" in six professional databases, such as Wiley, PubMed, ScienceDirect, ACS, Springer, and Web of Science, from 2000 to the present, summarizing the extraction, isolation, and purification methods for the four major categories (polysaccharides, proteins and peptides, saponins, and other components) of the A. japonicus chemical substances and 10 effective biological activities of A. japonicus. Included are anticoagulation, anticancer/antitumor activities, hematopoiesis, regulation of gut microbiota, and immune regulatory activities that correspond to traditional efficacy. Literature support is provided for the development of medicines and functional foods and related aspects that play a leading role in future directions.
Assuntos
Saponinas , Pepinos-do-Mar , Stichopus , Animais , Stichopus/química , Stichopus/fisiologia , Relação Estrutura-Atividade , AlimentosRESUMO
The main objective of this work was to characterize the acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) from the body wall of the sea cucumber scientifically called, Stichopus hermanni. For the extraction of ASC and PSC, the pre-treated sea cucumber body walls were subjected to 0.5 M acetic acid and 5 g L-1 pepsin, respectively. The yield of ASC (7.30% ± 0.30%) was found to be lower than the PSC (23.66% ± 0.15%), despite both ASC and PSC having similar chemical compositions except for the quantity of protein. The collagens produced from ASC and PSC show maximum peaks on ultraviolet-visible spectroscopic profiles at wavelengths of 230 and 235 nm, respectively, with no significant difference in the maximum temperature (Tmax ) of the extracted ASC and PSC. The ASC's coloration was whiter than that of the PSC. As a result, the collagen obtained from the body wall of the sea cucumber showed promise for usage as a substitute for collagen derived from marine sources. PRACTICAL APPLICATION: The two most popular methods of collagen extraction were acid hydrolysis and enzymatic hydrolysis. To determine whether the extracted collagen is a suitable substitute for animal collagen in different industries, it is required to characterize its physicochemical qualities. This study discovered a new application for marine collagen in the food industry: The sea cucumber has collagen with a greater yield in pepsin extraction with good physicochemical qualities.
Assuntos
Pepinos-do-Mar , Stichopus , Animais , Stichopus/química , Stichopus/metabolismo , Pepsina A/metabolismo , Pepinos-do-Mar/metabolismo , Colágeno/química , Ácidos/químicaRESUMO
To retard the protein degradation during sea cucumber processing, polyphenol extracts from Ascophyllum nodosum (PhE) was used as a potential antioxidant to maintain the structural integrity of sea cucumber body wall. Accordingly, the protection effects of PhE (0, 0.5, 1.0 and 1.5â¯mg PhE/g SFBW) against thermal degradation of the solid fragments of body wall (SFBW) have been investigated in order to evaluate their impact on the oxidation level and structural changes. Electronic Spin Resonance results showed that PhE could significantly inhibit the occurrence of oxidation by scavenging the free radicals. The effect of PhE on chemical analysis of soluble matters in SFBW was characterized by SDS-PAGE and HPLC. Compared with thermally treated SFBW, samples with PhE presented a decrease in protein dissolution. Thermal treatment resulted in the disintegration of collagen fibrils and fibril bundles in SFBW samples, while the density of collagen fibrils was increased, and the porosity decreased in samples with PhE. The results of FTIR and intrinsic tryptophan fluorescence confirmed that the structures of SFBW were modified by PhE. Besides, the denaturing temperature and decomposition temperature were both improved with the addition of PhE. These results suggested that PhE appeared to have a positive effect on lowering oxidation and improving thermostability and structural stability of SFBW, which could provide a theoretical basis for protecting sea cucumber body wall against degradation during thermal tenderization.
Assuntos
Ascophyllum , Pepinos-do-Mar , Stichopus , Animais , Pepinos-do-Mar/química , Stichopus/química , Stichopus/metabolismo , Polifenóis , Colágeno/químicaRESUMO
Major yolk proteins (MYPs), one class of the main abundant proteins in sea cucumber body wall, seem to garner more attention in recent years. Herein, a method using multiple reactions monitoring mass spectrometry (MRM-MS) was deliberatively developed to perform quantification analysis of three MYPs, i.e. BAH79576.1, BAH79577.1 and PIK45784.1. Contents of MYPs in body wall of fresh and dried sea cucumbers as well as in waste liquid of boiling and steaming were determined using their corresponding signature peptides of VDEFTGIVGSLR, KLDMYPPPLAR, LDMYPPPLAR, and SGHGEVMFVDSK. The loss of MYPs in the processing of sea cucumbers was directly verified by quantitation data of MYPs in sea cucumber body wall and the waste liquids. This study not only evidenced the loss of MYPs during the processing of sea cucumbers, but also implicated the potential of recycling MYPs from the processing waste water, providing helpful suggestions in maximizing the value of sea cucumbers.
Assuntos
Pepinos-do-Mar , Stichopus , Animais , Proteômica/métodos , Stichopus/química , Pepinos-do-Mar/metabolismo , Espectrometria de Massas , Proteínas do Ovo/metabolismoRESUMO
BACKGROUND: Apostichopus japonicus and Parastichopus californicus are two of the most important and profitable commercial sea cucumbers along the North Pacific coast. This study compared the body wall production rate (BWPR), proximate composition, amino acid, fatty acid, trace element and vitamin composition, and nonspecific immune enzyme activities of A. japonicus and P. californicus cultured in an artificial pond. RESULTS: The BWPR, crude fat and ash content in the body walls of A. japonicus and P. californicus showed remarkable differences (P < 0.05). For the 18 amino acids tested, differences in the contents of 15 were significant (P < 0.05) between the two species, except for threonine, methionine and histidine, and their first limiting amino acids were both methionine+cysteine. There were seven saturated and ten unsaturated fatty acids in their body walls, and except for 18:1 and 20:1, the content differences of the other 15 fatty acids were all significant (P < 0.05). Furthermore, between the two sea cucumbers, differences in the content of seven trace elements (Cu, Fe, Mn, Zn, Cr, Ni, Se) and six vitamins (B1, B3, B5, B9, C, E) were significant (P < 0.05). The activities of superoxide dismutase (SOD), catalase (CAT), acid phosphatase (ACP) and alkaline phosphatase (AKP) also showed distinct differences (P < 0.05). CONCLUSION: There are greater differences in the biochemical compositions and contents between A. japonicus and P. californicus, each with its own unique quality advantages. A. japonicus and P. californicus have high nutritional value, which are both the superior sea cucumbers. © 2022 Society of Chemical Industry.
Assuntos
Pepinos-do-Mar , Stichopus , Aminoácidos/metabolismo , Animais , Ácidos Graxos/análise , Metionina/metabolismo , Valor Nutritivo , Pepinos-do-Mar/química , Stichopus/químicaRESUMO
BACKGROUND: Zinc absorption in intestinal system could be strongly affected by the gastrointestinal digestion and absorption of zinc-chelating peptides serving as zinc carriers. In this study, a novel zinc-chelating sea cucumber synthetic peptide (SCSP) was synthesized to estimate its gastrointestinal digestion and promotive effect of zinc absorption in vitro. RESULTS: Analysis of isothermal titration calorimetry suggested that the binding of SCSP and zinc (N ≈ 1) was exothermic, with relatively weak binding affinity (K = 1.0 × 10-3 mol L-1 ). The formation of SCSP-Zn complexes brought morphological changes to the peptides confirmed by scanning electron microscopy (SEM), which also indicated 6.88% of the existence of zinc element. In addition, the SCSP-Zn complexes remained stable under simulated human gastrointestinal digestion. In an in vitro study, the SCSP-Zn complex could successfully transport through the intestinal membrane in the model of everted rat gut sacs (nearly 7.5 µM cm-2 ) as well as Caco-2 cells where the zinc transport reached 0.0014 mg mL-1 carried by SCSP. Fluorescence staining experiments revealed free zinc accumulation inside the tissues and cells treated with the SCSP-Zn complex. CONCLUSIONS: The chelation SCSP-Zn had the promotion ability of zinc absorption in vitro and ex vivo experiments, which suggested a theoretical basis for the design and production of effective zinc chelating peptides as zinc carriers to improve zinc bioavailability. © 2022 Society of Chemical Industry.
Assuntos
Pepinos-do-Mar , Stichopus , Animais , Células CACO-2 , Digestão , Humanos , Peptídeos/química , Ratos , Pepinos-do-Mar/química , Stichopus/química , Zinco/metabolismoRESUMO
Sea cucumber Stichopus chloronotus is a traditional tonic food with high nutritive value in Southern China. Fucoidan from sea cucumber Stichopus chloronotus (Fuc-Sc) is its main bio-active polysaccharide, the immune-activation effects of which have been fully investigated on RAW264.7 cells in the present study. The results indicated that Fuc-Sc could stimulate the RAW264.7 cells by promoting the production of NO, TNF-α, IL-6 and IL-10. Western blot and RT-PCR analysis revealed that TLR4 and TLR2 were involved in the recognition of Fuc-Sc and activation of downstream NF-κB signal pathway. Moreover, the chemical structure parameter molecular weight showed obvious impact on the stimulation effects of Fuc-Sc on NO production. Degraded product of Fuc-Sc with weight average molecular weight of 113.1 × 104 Da exhibited higher activities than that of intact Fuc-Sc, suggesting the existent of optimum chain length to exert its highest activities. Taken together, Fuc-Sc exerted its immunostimulating activity via TLR2/4 activation of NF-κB pathway and showed potentials to be a good immunoadjuvant.
Assuntos
Adjuvantes Imunológicos/farmacologia , NF-kappa B/metabolismo , Polissacarídeos/farmacologia , Stichopus/química , Receptor 2 Toll-Like/metabolismo , Adjuvantes Imunológicos/química , Animais , Sobrevivência Celular/efeitos dos fármacos , China , Humanos , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Camundongos , Peso Molecular , Óxido Nítrico/metabolismo , Polissacarídeos/química , Células RAW 264.7 , Pepinos-do-Mar/química , Transdução de Sinais/efeitos dos fármacos , Relação Estrutura-Atividade , Receptor 4 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
SCOPE: This study aims to investigate the protective effect of Apostichopus japonicus oligopeptide (AJOP) on hyperuricemia, demonstrate the modulation of the gastrointestinal tract (GIT) microbiota, and clarify the underlying microbiota-dependent mechanism. METHODS AND RESULTS: Hyperuricemic mice treated with AJOP and subjected to corresponding fecal microbiota transplantation (FMT) are used to observe the beneficial effects of AJOP and microbiota. Gene transcriptions are measured using quantitative real-time PCR. The GIT (stomach, colon, cecum, and feces) microbiota is analyzed by 16S rDNA sequencing and the short-chain fatty acids are detected using GC-MS. Dietary administration of AJOP significantly alleviates hyperuricemia, regulates uric acid metabolism, inhibites the activation of the NLRP3 inflammasome and NF-κB-related signaling pathway, and restores m6A methylation levels. In addition, substantial heterogeneity is observed in GIT microbiota. Furthermore, FMT effectively alleviates hyperuricemia in mice by selectively regulating the corresponding pathways associated with AJOP treatment, indicating that the mechanism underlying the protective effects of AJOP is partly microbiota-dependent. CONCLUSION: This study demonstrates that AJOP exerts a protective effect on hyperuricemic mice by regulating uric acid metabolism, resulting in substantial heterogeneity among the GIT microbiota, thus mediating the beneficial effects in a microbiota-dependent manner.
Assuntos
Microbioma Gastrointestinal/efeitos dos fármacos , Hiperuricemia/tratamento farmacológico , Oligopeptídeos/farmacologia , Preparações de Plantas/farmacologia , Stichopus/química , Animais , Transplante de Microbiota Fecal , Masculino , Camundongos , Camundongos Endogâmicos ICR , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Ácido Úrico/metabolismoRESUMO
A fucosylated chondroitin sulfate was isolated from the body wall of sea cucumber Stichopus japonicus (FCSsj), whose structure was characterized by NMR spectroscopy and HILIC-FTMS. At the ratio of 1.00:0.26:0.65, three fucosyl residues were found: 2,4-disulfated-fucose (Fuc2,4S), 4-sulfated-fucose (Fuc4S) and 3,4-disulfated-fucose (Fuc3,4S), which were only linked to the O-3 of glucuronic acid residues (GlcA). Besides mono-fucosyl moieties, di-fucosyl branches, namely Fuc2,4Sα(1â3)Fuc4S, were also found to be attached to the O-3 of GlcA. The antidiabetic activity of FCSsj was evaluated using glucosamine induced insulin resistant (IR) Hep G2 cells in vitro. It was found that FCSsj significantly promoted the glucose uptake and glucose consumption of IR-Hep G2 cells in a dose-dependent manner, and could alleviate the cell damage. Furthermore, FCSsj could promote the glycogen synthesis in the glucosamine-induced IR-Hep G2 cells. These results provided a supplement for studying the antidiabetic activity of FCSsj.
Assuntos
Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacologia , Hipoglicemiantes/química , Hipoglicemiantes/farmacologia , Stichopus/química , Animais , Fucose/química , Glucose/metabolismo , Ácido Glucurônico/química , Glicogênio/metabolismo , Células Hep G2 , Humanos , Resistência à Insulina , Espectroscopia de Ressonância Magnética/métodos , Pepinos-do-Mar/químicaRESUMO
The sea cucumber, Apostichopus japonicas, is an important economic species with high nutritive value. In recent years, driven by significant market demand, the sea cucumber breeding industry has developed rapidly. Body color and number of papillae are important factors that determine the value and price of sea cucumbers. In this study, metabolite profiling of four sea cucumber varieties (green, white, purple and spiny) was performed using ultra-performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry (UPLC-QTOF-MS) combined with multivariate analysis. Orthogonal partial least-squares discriminant analysis (OPLS-DA) clearly discriminated the body wall metabolites of four sea cucumber varieties. Differential metabolites included fatty acids, phospholipids, and sugars. KEGG metabolic pathway analysis revealed that carbohydrate, protein, and phospholipid metabolism were highly conserved among the varieties. These results provide a comprehensive insight into differences in the metabolite profile of four A. japonicus varieties and a deeper understanding of sea cucumber varieties breeding.
Assuntos
Metabolômica , Stichopus/metabolismo , Animais , Cromatografia Líquida , Análise Discriminante , Stichopus/químicaRESUMO
Marine sulfated polysaccharides have aroused widespread concern for their various structures and bioactivities. Peroxide depolymerization is a common strategy in analysis of structures and structure-activity relationships of polysaccharides. However, confirming the depolymerization process and exact structures of the degradation products is still a considerable challenge. This study reported the structures of a fucan sulfate (FS) from sea cucumber Stichopus herrmanni and its depolymerized products (dFS) prepared by peroxide degradation. The FS was elucidated with a highly regular structure, {-3)-L-Fuc2S-(α1-}n. Structure analysis of oligosaccharides purified from dFS suggested that peroxide degradation involved in cleavage of glycosidic bonds and oxidative modification of reducing end of sugar residue, while no break in sugar ring was observed. Both FS and series of dFSs exhibited significant anticoagulant activities due to their anti-thrombin effects in presence of heparin cofactor II and their potencies were related to their molecular sizes, dFS with â¼ 20 kDa showed the strongest activity.
Assuntos
Anticoagulantes/química , Coagulação Sanguínea/efeitos dos fármacos , Peróxidos/química , Polissacarídeos/química , Stichopus/química , Animais , Anticoagulantes/isolamento & purificação , Anticoagulantes/farmacologia , Organismos Aquáticos , Testes de Coagulação Sanguínea , Sequência de Carboidratos , Cofator II da Heparina/farmacologia , Humanos , Hidrólise , Peso Molecular , Polissacarídeos/isolamento & purificação , Polissacarídeos/farmacologia , Stichopus/fisiologia , Relação Estrutura-Atividade , Trombina/antagonistas & inibidores , Trombina/metabolismoRESUMO
Sea cucumbers were nutritional food and traditional Chinese medicine. In this study, fucosylated chondroitin sulfate from sea cucumber Stichopus chloronotus (fCS-Sc), a potential anticoagulant agent and immunological adjuvant, was investigated for its immune activation effects on RAW 264.7 macrophage for the first time. The results indicated that fCS-Sc could significantly promote the proliferation, the pinocytic activity of RAW 264.7 cells, and the production of NO, TNF-α, IL-1ß, and IL-6. The fluorescence labeling assay indicated that fCS-Sc could bind to the macrophage. Moreover, the specific pattern recognition receptor inhibition assays showed that toll-like receptor 4 (TLR4) and TLR2 were involved in the recognition of fCS-Sc. Western blot assays indicated that fCS-Sc could induce degradation of cytoplasm IκB-α, and promotion of NF-κB p65 subunit translocation to nucleus, leading to a functional improvement of macrophage through NF-κB pathway. The results suggested that fCS-Sc might served as a promising candidate of immunomodulator.