RESUMEN
Ginger polysaccharides (GP) promote growth and development in fish. However, the effects of GP on crucian carp remain unclear. The present study investigated the effects of GP on the growth performance, immunity, intestinal microbiota, and disease resistance in crucian carp. Four treatment groups were established with different concentrations of GP (0.1 %, 0.2 %, 0.4 %, and 0.8 %). GP was not added as the control group, and the feeding period lasted for 56 d, followed by a 96-h anti-infection treatment using Aeromonas hydrophila. The results showed that dietary GP significantly improved growth performance, especially in the 0.4 % GP group. Furthermore, GP administration notably increased serum lysozyme (LMZ) activity, digestive enzyme performance, and antioxidant capacity of crucian carp. Moreover, dietary inclusion of GP up-regulated the expression of tumour necrosis factor-α (TNF-α), interleukin-8 (IL-8), interferon-γ (IFN-γ), and nuclear factor kappa-B (NF-κB) genes while down-regulating IL-10 and transforming growth factor-ß (TGF-ß) gene expressions, thus promoting liver health in crucian carp. Additionally, incorporating GP into the diet regulated both the diversity and composition of the intestinal microbiota in crucian carp, explicitly enhancing the relative abundance of beneficial bacteria, such as Fusobacteriota and Firmicutes. Therefore, GP reduces the mortality of crucian carp infected with A. hydrophila. In conclusion, this study provides novel insights into the application of dietary GP in cultured fish and evaluates the value of traditional Chinese medicinal polysaccharides against pathogenic bacteria.
Asunto(s)
Aeromonas hydrophila , Antioxidantes , Carpas , Resistencia a la Enfermedad , Enfermedades de los Peces , Microbioma Gastrointestinal , Polisacáridos , Zingiber officinale , Animales , Aeromonas hydrophila/efectos de los fármacos , Microbioma Gastrointestinal/efectos de los fármacos , Polisacáridos/farmacología , Polisacáridos/química , Resistencia a la Enfermedad/efectos de los fármacos , Antioxidantes/farmacología , Enfermedades de los Peces/microbiología , Enfermedades de los Peces/inmunología , Zingiber officinale/química , Carpas/crecimiento & desarrollo , Carpas/inmunología , Carpas/microbiología , Infecciones por Bacterias Gramnegativas/veterinaria , Infecciones por Bacterias Gramnegativas/inmunología , Infecciones por Bacterias Gramnegativas/tratamiento farmacológico , Suplementos Dietéticos , Alimentación AnimalRESUMEN
In this study, the ginger polysaccharides extracted from hot water (HW-G) were modified with subcritical water (SW-G) to effectively regulate their immune activity, and the relationship between polysaccharide chain conformation and immune activity at different subcritical water temperatures was investigated. The results indicated that, compared with HW-G, the xylose and mannose were degraded at high temperatures. The molecular weight of ginger polysaccharide decreased from 1.083 × 106 g/mol to 3.113 × 105 g/mol after subcritical water modification (100-160 °C). The chain conformation transitioned from rigid rod chain to semi-rigid chain and eventually to random coil. The degree of relaxation of the polysaccharide chains showed a continuous increase trend. Additionally, ginger polysaccharide modified by subcritical water at 130 °C was found to promote the proliferation and phagocytosis of 264.7 cells more obviously and signally increase the secretion levels of NO, IL-6, TNF-α and IL-1ß. When the subcritical water temperature exceeds 130 °C, the activity of ginger polysaccharide begins to decline rapidly. These findings demonstrate a close correlation between polysaccharide chain conformation and immunomodulatory activity, confirming the feasibility of the subcritical water temperature effect as a means of immune activity regulation, which opens up a new approach to obtaining highly active polysaccharides.
Asunto(s)
Agua , Zingiber officinale , Temperatura , Polisacáridos/farmacología , AntioxidantesRESUMEN
Alkaline stress poses a significant challenge to the healthy growth of fish. Ginger polysaccharide (GP) is one of the main active substances in ginger and has pharmacological effects, such as anti-oxidation and immune regulation. However, the physiological regulatory mechanism of GP addition to diet on alkalinity stress in crucian carp remains unclear. This study aimed to investigate the potential protective effects of dietary GP on antioxidant capacity, gene expression levels, intestinal microbiome, and metabolomics of crucian carp exposed to carbonate (NaHCO3). The CK group (no GP supplementation) and COG group (NaHCO3 stress and no GP supplementation) were set up. The GPCS group (NaHCO3 stress and 0.4% GP supplementation) was stressed for seven days. Based on these data, GP significantly increased the activities of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), acid phosphatase (ACP), and alkaline phosphatase (AKP) in carp under alkalinity stress (p < 0.05) and decreased the activity of malon dialdehyde (MDA) (p < 0.05). GP restored the activity of GSH-PX, ACP, and AKP to CK levels. The expression levels of tumor necrosis factor ß (TGF-ß), tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), and interleukin 8 (IL-8) genes were decreased, and the expression levels of determination factor kappa-B (NF-κB) and interleukin 10 (IL-10) genes were increased (p < 0.05). Based on 16â¯S rRNA high-throughput sequencing, GP improved the changes in the intestinal microbial diversity and structural composition of crucian carp caused by NaHCO3 exposure. In particular, GP increased the relative abundance of Proteobacteria and Bacteroidetes and decreased the relative abundance of Actinobacteria. The metabolic response of GP to NaHCO3 exposed crucian carp guts was studied using LC/MS. Compared to the COG group, the GPCS group had 64 different metabolites and enriched 10 metabolic pathways, including lipid metabolism, nucleotide metabolism, and carbohydrate metabolism. The addition of GP to feed can promote galactose metabolism and provide an energy supply to crucian carp, thus alleviating the damage induced by alkalinity stress. In conclusion, GP can mitigate the effects of NaHCO3 alkalinity stress by regulating immune function, intestinal flora, and intestinal metabolism in crucian carp. These findings provide a novel idea for studying the mechanism of salt-alkali tolerance in crucian carp by adding GP to feed.
Asunto(s)
Carpas , Microbioma Gastrointestinal , Zingiber officinale , Animales , Carpa Dorada/metabolismo , Carpas/metabolismo , Antioxidantes/metabolismo , Dieta , Carbonatos , Alimentación Animal/análisisRESUMEN
Recently, targeting myeloid-derived suppressor cells (MDSCs) which mainly play an immunosuppressive role in tumor microenvironment has become a hot spot in tumor immunotherapy. This study focuses on biological effect of ginger polysaccharide extracted from natural plants on promoting apoptosis of MDSCs by regulating lipid metabolism. An MTT assay was used to detect the inhibitory effect of ginger polysaccharide on the growth of an MDSC-like cell line (MSC-2). The apoptosis-promoting effect of ginger polysaccharide on MSC-2 cells was detected by flow cytometry. Expression levels of apoptosis proteins (caspase 9 and Bcl-2) and lipid metabolism enzymes (fatty acid synthase (FASN) and diacylglycerol acyltransferase 2) in MSC-2 cells treated with different concentrations of ginger polysaccharide were detected by western blot assay. Nile red staining was used to quantitatively detect the effect of ginger polysaccharide on lipid droplet synthesis. Ginger polysaccharide inhibited proliferation of MSC-2 cells and promoted their apoptosis by upregulating pro-apoptotic caspase 9 protein, downregulating anti-apoptotic Bcl-2 protein, inhibiting expression of FASN and diacylglycerol acyltransferase 2 (key enzymes in fatty acid synthesis and lipid droplet formation, respectively). Ginger polysaccharide promoted apoptosis of MDSCs by regulating key lipid metabolism enzymes, inhibiting fatty acid synthesis and lipid droplet accumulation, and reducing the energy supply of cells.
Asunto(s)
Células Supresoras de Origen Mieloide , Zingiber officinale , Caspasa 9/metabolismo , Metabolismo de los Lípidos , Diacilglicerol O-Acetiltransferasa/metabolismo , Diacilglicerol O-Acetiltransferasa/farmacología , Apoptosis , Proteínas Reguladoras de la Apoptosis/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Polisacáridos/farmacología , Polisacáridos/metabolismo , Ácidos Grasos/farmacología , Proliferación CelularRESUMEN
A neutral ginger polysaccharide fraction (NGP) was isolated from ginger, and its primary structures and immunomodulatory activity were investigated. The results showed that NGP had a low molecular weight of 6305 Da. NGP principally consisted of glucose (93.88%) together with minor levels of galactose (3.27%) and arabinose (1.67%). Besides, results of methylation analysis and 1D/2D NMR spectroscopy demonstrated that NGP was α-glucan which had the main chain of 1,4-linked α-d-Glcp and α-d-Glcp residues branched at C-6 position which was different from the common triple helical ß-glucans. NGP also displayed a remarkable immunological activity on the RAW264.7 cells in vitro. It could significantly enhance the proliferation of macrophages without cytotoxicity and increase the production of immune substances (NO, TNF-α, IL-1ß and IL-6). The secretion at the concentration of 200 µg/mL was 29.41 µM, 1496.71, 44.30 and 1889.83 pg/mL for each substance, respectively. The results indicated that NGP could be a potential immune agent and might provide meaningful information for further chain conformation and immune mechanism research.
Asunto(s)
Factores Inmunológicos/química , Factores Inmunológicos/farmacología , Polisacáridos/química , Polisacáridos/farmacología , Zingiber officinale/química , Animales , Factores Inmunológicos/análisis , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Metilación , Ratones , Peso Molecular , Polisacáridos/análisis , Células RAW 264.7RESUMEN
In this study, ginger polysaccharide (GP), ginger polysaccharide 1 (GP1), and ginger polysaccharide 2 (GP2) from ginger were firstly modified by sulfation. Fourier transform infrared, and nuclear magnetic resonance spectra investigation of sulfated ginger polysaccharide (SGP), sulfated ginger polysaccharide 1 (SGP1), and sulfated ginger polysaccharide 2 (SGP2) revealed that the sulfation successfully occurred with the characteristic absorption peak of polysaccharide. Congo red experiment showed that triple helical structure existed in SGP and SGP1, but random coils existed in SGP2. SGP, SGP1, and SGP2 all showed a rough and rugged surface with plenty of small pores. The blood clotting time of SGP2 or SGP at 2 mg/mL in activated partial thromboplastin time (APTT) assay was 41.42 or 38.01 s, respectively, which were approximately 1.33- and 1.22-fold longer than that of the physiological saline. Compared to the saline control group, prothrombin time (PT) was increased by 1.22-fold with the addition of GP at 2 mg/mL. However, no clotting inhibition phenomenon was observed in thrombin time test even at the concentrations that APTT and PT were obviously prolonged. It indicated that GP2, SGP2, and SGP inhibited the intrinsic pathway of coagulation, but GP inhibited both the intrinsic and extrinsic pathways of coagulation. Hence, ginger polysaccharides might be used as anticoagulants and therapeutic reagents for thrombosis.
Asunto(s)
Anticoagulantes/farmacología , Polisacáridos/química , Polisacáridos/farmacología , Sulfatos/química , Zingiber officinale/química , Coagulación Sanguínea/efectos de los fármacos , Carbohidratos de la Dieta/farmacología , Humanos , Espectroscopía de Resonancia Magnética , Tiempo de Tromboplastina Parcial , Tiempo de Protrombina , Tiempo de TrombinaRESUMEN
Three different extraction technologies including hot water extraction (HWE), enzyme assisted extraction (EAE) and ultrasonic cell grinder extraction (UCGE) were employed to extract crude ginger polysaccharides (GPs) under their respective best parameters, then crude GPs were purified by DEAE cellulose-52 and Sephadex G-200 size-exclusion chromatography in that order. Five GPs fractions (HGP, EGP1, EGP2, UGP1, and UGP2, respectively) were obtained. The differences of five GPs in chemical composition, characterization and antitumor activities were further compared. The molecular weights were different in five GPs, varying from 11.81 to 1831.75 kDa. Mannose and glucose as the main monosaccharide and the glycosidic linkage of â4)-α-D-Glc(1â and -α-Manp-(1â existed in both five GPs. While EGP2 and UGP1 possessed specific structure of â6)-ß-D-Galp-(1â and UGP1 contained more sulfate group. Moreover, UGP1 exhibited strong inhibitory effect on three tumor cells especially the colon cancer. The inhibition rates of UGP1 on H1975, HCT116 and MCF-7 were 23.339 ± 2.285%, 56.843 ± 2.405% and 21.061 ± 1.920% respectively. The study indicated GPs extracted by UCGE could reserve more active structure and inhibit colon cancer more significantly.
Asunto(s)
Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Fraccionamiento Químico/métodos , Polisacáridos/aislamiento & purificación , Polisacáridos/farmacología , Zingiber officinale/química , Antineoplásicos/química , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Humanos , Polisacáridos/químicaRESUMEN
In this study, ginger polysaccharide (GP) was obtained from ginger by enzymatic method, its chemical properties and antitumor activity were investigated. The results indicated that the composition and proportion of GP were lrhamnose, darabinose, dmannose, dglucose and dgalactose in a molar ratio of 3.64:5.37:3.04:61.03:26.91, GP had the characteristic absorption peak of polysaccharide. Congo red experiment showed that GP had a triple helix structure, which could have anti-tumor effect. Furthermore, MTT assay, cell morphology observation, nuclear morphology observation and reactive oxygen species observation demonstrated that GP had significant antitumor effect. Flow cytometry suggested that GP could promote apoptosis and arrest cells in G0-G1 phase. Real-time fluorescence quantification and Western blot revealed that GP could up-regulate the expression of Bax, Fas, FasL, caspase-3, p21 and p53, and down-regulate the expression of Bcl-2. These studies suggested that GP would be used as an antitumor drug in foods to promote the development of functional foods.
Asunto(s)
Apoptosis/efectos de los fármacos , Carcinoma Hepatocelular/tratamiento farmacológico , Puntos de Control del Ciclo Celular/efectos de los fármacos , Neoplasias Hepáticas/tratamiento farmacológico , Polisacáridos/farmacología , Zingiber officinale/química , Proteínas Reguladoras de la Apoptosis/metabolismo , Carcinoma Hepatocelular/metabolismo , Línea Celular Tumoral , Regulación hacia Abajo/efectos de los fármacos , Fase G1/efectos de los fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Fase de Descanso del Ciclo Celular/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Two components ginger polysaccharide 1 (GP1) and ginger polysaccharide 2 (GP2) were extracted. The results showed that the molecular weights of GP1 and GP2 were 6128â¯Da and 12,619â¯Da, respectively. The composition and proportion of GP1 and GP2 were mannose, glucose and galactose in a molar ratio of 4.96: 92.24: 2.80 and arabinose, mannose, glucose and galactose in a molar ratio of 4.78: 16.70: 61.77: 16.75, respectively, illustrating that GP1 and GP2 were not a kind of homopolysaccharide. GP1 has a three-helix structure, and the structure is closely linked. GP2 contains sulfuric acid groups, and has a high oxidation resistance, its structure is more evacuated and messy.