Inulin-type fructan CP-A,a predominant polysaccharide in Codonopsis pilosula,demonstrates regulatory effects on immune activity and anti-inflammation.The efficacy of CP-A in treating ulcerative colitis(UC)is,however,n...Inulin-type fructan CP-A,a predominant polysaccharide in Codonopsis pilosula,demonstrates regulatory effects on immune activity and anti-inflammation.The efficacy of CP-A in treating ulcerative colitis(UC)is,however,not well-established.This study employed an in vitro lipopolysaccharide(LPS)-induced colonic epithelial cell model(NCM460)and an in vivo dextran sulfate sodium(DSS)-induced colitis mouse model to explore CP-A’s protective effects against experimental colitis and its underlying mechanisms.We monitored the clinical symptoms in mice using various parameters:body weight,disease activity index(DAI),colon length,spleen weight,and histopathological scores.Additionally,molecular markers were assessed through enzyme-linked immunosorbent assay(ELISA),quantitative real-time polymerase chain reaction(qRT-PCR),immunofluorescence(IF),immunohistochemistry(IHC),and Western blotting assays.Results showed that CP-A significantly reduced reactive oxygen species(ROS),tumor necrosis factor-alpha(TNF-α),and interleukins(IL-6,IL-1β,IL-18)in LPS-induced cells while increasing IL-4 and IL-10 levels and enhancing the expression of Claudin-1,ZO-1,and occludin proteins in NCM460 cells.Correspondingly,in vivo findings revealed that CPA administration markedly improved DAI,reduced colon shortening,and decreased the production of myeloperoxidase(MPO),malondialdehyde(MDA),ROS,IL-1β,IL-18,and NOD-like receptor protein 3(NLRP3)inflammasome-associated genes/proteins in UC mice.CP-A treatment also elevated glutathione(GSH)and superoxide dismutase(SOD)levels,stimulated autophagy(LC3B,P62,Beclin-1,and ATG5),and reinforced Claudin-1 and ZO-1 expression,thereby aiding in intestinal epithelial barrier repair in colitis mice.Notably,the inhibition of autophagy via chloroquine(CQ)diminished CP-A’s protective impact against colitis in vivo.These findings elucidate that CP-A’s therapeutic effect on experimental colitis possibly involves mitigating intestinal inflammation through autophagymediated NLRP3 inflammasome inactivation.Consequently,inulin-type fructan CP-A emerges as a promising drug candidate for UC treatment.展开更多
To develop an HPLC-ELSD method for the determination of nine inulin-type fructo-oligosaccharides in inulin,the HPLC-ELSD system consisted of Waters XBridge■ Amide column(4.6 mm×250 mm,5μm)with a gradient elutio...To develop an HPLC-ELSD method for the determination of nine inulin-type fructo-oligosaccharides in inulin,the HPLC-ELSD system consisted of Waters XBridge■ Amide column(4.6 mm×250 mm,5μm)with a gradient elution mobile phase consisting of acetonitrile and water at a flow rate of 1.2 mL/min at 30°C.The detector was an Agilent Technologies 380-ELSD.The drift tube temperature for the ELSD was set at 55°C with a nitrogen flow rate of 1.8 L/min.The injection volume was 15μL.The results showed that the detection range for the nine inulin-type fructo-oligosaccharides was 3.81–30.60μg R^(2)=0.99969 for kestose,3.73–29.97μg R^(2)=0.99981 for nystose,3.82–30.69μg R^(2)=0.99993 for fructosylnystose,3.80–30.48μg R^(2)=0.99995 for GF5,3.73–29.96μg R^(2)=0.99993 for GF6,3.78–30.30μg R^(2)=0.99983 for GF7,3.82–30μg R^(2)=0.99989 for GF8,3.71–29.80μg R^(2)=0.99974 for GF9,3.61–29.00μg R^(2)=0.99970 for GF10,respectively.The recovery of the nine oligosaccharides ranged between 96.48%–100.84%(n=6).The method was simple,accurate,and reproducible that it could be used as an analytical method for evaluating the quality of inulin effectively.展开更多
Fructan is not only a carbon source for storage but also plays an important role as anti-stress agents in many plant species. Complex fructans having both β-（2,1）- and β-（2,6）-linked fructosyl units accumulate i...Fructan is not only a carbon source for storage but also plays an important role as anti-stress agents in many plant species. Complex fructans having both β-（2,1）- and β-（2,6）-linked fructosyl units accumulate in Triticeae plants commonly. Three enzymes （sucrose： sucrose 1-fructosyltransferase, 1-SST, EC： 2.4.1.99; sucrose： fructan 6-fructosyltransferase, 6- SFT, EC： 2.4.1.10; and fructan： fructan 1-fructosyltransferase, 1-FFT, EC： 2.4.1.100） were involved in fructan biosynthesis in Triticeae plant species. We successfully isolated these genes from tetraploid wheat （Triticum turgidum, genotype： AABB）, common wheat （Triticum aestivum L., genotype： AABBDD） and three wild relatives of common wheat, Triticum urartu Thum. （the origin of the AA genome）, Aegilops speltoides （Tausch） Gren. （the putative source of the SS genome） and Aegilops tauschii Coss. （the source of the DD genome）. Sequence analysis revealed that all the FBEs （fructan biosynthetic enzymes） had three highly conserved functional motifs except 1-SST （EU981912） from tetraploid wheat species only with conserved DPNG. Low pI （isoelectric point） and potential N-glycosylation sites were predicted, which were crucial for protein compartmentation and post-translational process. Analysis on subcelluar localization signals showed that only 6-SFT had vacuolar-directed signal. Sequences alignment result showed that 1-SST and 1-FFT were more conservative and had closer relationship each other, while 6-SFT was more active during the evolution processing. According to the syntenic relationship between wheat and rice genome, FBEs were predicated to be located on the homeologous group 6 and group 2 chromosomes. Expression profile confirmed that expression of all the three FBEs were drought-stress induced. This study can assist to establish a useful theoretical platform for cold- or drought-tolerant improvement of wheat by modulating FBEs expression.展开更多
Translocation of previously accumulated nitrogen and carbohydrates from vegetative tissue of the wheat plant is a major assimilate source for grain filling. This study was conducted to examine genotype differences in ...Translocation of previously accumulated nitrogen and carbohydrates from vegetative tissue of the wheat plant is a major assimilate source for grain filling. This study was conducted to examine genotype differences in nitrogen and fructan translocation and their relationships to grain yield and protein content. Effects indicated that significant genotype differences existed for nitrogen accumulation at anthesis and fructan at milk stage and their translocation. Two high protein genotypes, Cunningham and PST90-19, accumulated more nitrogen before anthesis and had greater nitrogen translocation, but lower post-anthesis nitrogen uptake, than two low protein genotypes, SUN109A and TM56. Among plant parts, leaves were the major storage for tissue nitrogen and provided the overwhelming proportion of the total nitrogen translocation, whereas for fructan accumulation and translocation it was the stems. The two high protein genotypes had a higher percentage of their grain nitrogen derived from nitrogen translocation, while for the two low protein ones, it was from post-anthesis nitrogen uptake and assimilation. Increasing nitrogen application increased nitrogen accumulation and translocation, but decreased fructan accumulation and translocation. High grain protein content was associated with high nitrogen translocation from leaves, stems and the total plant, while high grain yield was related to high fructan translocation from stems and the total plant. Fructan translocation was negatively correlated to grain protein content. Nitrogen and fructan translocation were not correlated with each other.展开更多
The aim of the study was to contribute to the information on agave soluble fibers since research has been focused on chicory fiber but not in agave products;thus we assess the acute toxicity and genotoxicity of two or...The aim of the study was to contribute to the information on agave soluble fibers since research has been focused on chicory fiber but not in agave products;thus we assess the acute toxicity and genotoxicity of two organic and high purity dietary soluble fibers from agave, Metlin? and Metlos?. We performed an acute toxicity assay in Hsd:ICR mice and Hsd:Wistar rats and an in vivo genotoxic test. Results showed that there are no deaths at any doses or genotoxicity, so it can be concluded that these products are non-toxic, at the administrated doses and none showed a cytotoxic, clastogenic or aneuploidic effect.展开更多
Fructans are important in the survival of plants and also valuable for humans as potentially health promoting food ingredients. In this study fructan content and composition were determined in grains of 20 barley bree...Fructans are important in the survival of plants and also valuable for humans as potentially health promoting food ingredients. In this study fructan content and composition were determined in grains of 20 barley breeding lines and cultivars with a wide variation in chemical composition, morphology and country of origin, grown at one site in Chile. There was significant genotypic variation in grain fructan content ranging from 0.9% to 4.2% of grain dry weight. Fructan degree of polymerisation (DP) was analysed using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Changes in the distribution of different chain lengths and the pattern of structures of fructan were detected with increasing amount of fructan in the different barleys. A positive correlation was found between fructan content and the relative amount of long chain fructan (DP > 9) (r = 0.54, p = 0.021). Our results provide a basis for selecting promising barley lines and cultivars for further research on fructan in barley breeding with the aim to produce healthy food products.展开更多
为探究乳酸菌果聚糖代谢机制和开发低果聚糖发酵食品,本研究对287株酸面团来源的乳酸菌进行果聚糖利用能力筛选,获得1株嗜果聚糖乳酸菌19M03,之后利用高通量测序和生物信息学技术对该菌株进行全基因组测序和果聚糖代谢机制分析。测序结...为探究乳酸菌果聚糖代谢机制和开发低果聚糖发酵食品,本研究对287株酸面团来源的乳酸菌进行果聚糖利用能力筛选,获得1株嗜果聚糖乳酸菌19M03,之后利用高通量测序和生物信息学技术对该菌株进行全基因组测序和果聚糖代谢机制分析。测序结果表明,菌株19M03基因组含有1个染色体和9个质粒,基因组全长3246316 bp,GC相对含量44.53%,包含3084个编码基因、66个tRNA和16个rRNA。系统发育关系分析表明,分类学上菌株19M03属于植物乳植杆菌。基因功能注释结果表明,在同源蛋白簇、基因本体论、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)和碳水化合物活性酶数据库中,19M03共编码357个碳水化合物代谢相关基因和351个碳水化合物转运相关基因,其中与果聚糖代谢和转运相关的基因均有12个。根据这些基因在KEGG数据库中的代谢关系,提出19M03果聚糖代谢模型。该模型解析了果聚糖水解过程及其催化反应的细胞定位(既可胞内,又可胞外)、细胞膜中运输低聚果糖和果糖的特异转运蛋白(IIABC^(FOS)、IIABCD^(fru)、IIABC^(fru))、细胞内果糖分解代谢途径(n=4)和果糖合成代谢途径(n=1)及其所有相关酶的编码基因。此外,耐药和毒力基因分析证明19M03具有良好的安全性。总之,本研究筛选获得了1株嗜果聚糖乳酸菌19M03,并在全基因组水平揭示了19M03果聚糖代谢机制,这些结果可为阐明乳酸菌果聚糖代谢机制提供重要的理论参考,为今后低果聚糖功能食品的开发和应用提供优质的菌种资源。展开更多
基金supported by the National Natural Science Foundation of China(No.81904031)National Key Research and Development Program of China(No.2019YFC1710800)+1 种基金the Natural Science Foundation of Shanxi Province(No.201901D211325)the Science Research Start-up Fund for Doctor of Shanxi Medical University(No.XD1802).
文摘Inulin-type fructan CP-A,a predominant polysaccharide in Codonopsis pilosula,demonstrates regulatory effects on immune activity and anti-inflammation.The efficacy of CP-A in treating ulcerative colitis(UC)is,however,not well-established.This study employed an in vitro lipopolysaccharide(LPS)-induced colonic epithelial cell model(NCM460)and an in vivo dextran sulfate sodium(DSS)-induced colitis mouse model to explore CP-A’s protective effects against experimental colitis and its underlying mechanisms.We monitored the clinical symptoms in mice using various parameters:body weight,disease activity index(DAI),colon length,spleen weight,and histopathological scores.Additionally,molecular markers were assessed through enzyme-linked immunosorbent assay(ELISA),quantitative real-time polymerase chain reaction(qRT-PCR),immunofluorescence(IF),immunohistochemistry(IHC),and Western blotting assays.Results showed that CP-A significantly reduced reactive oxygen species(ROS),tumor necrosis factor-alpha(TNF-α),and interleukins(IL-6,IL-1β,IL-18)in LPS-induced cells while increasing IL-4 and IL-10 levels and enhancing the expression of Claudin-1,ZO-1,and occludin proteins in NCM460 cells.Correspondingly,in vivo findings revealed that CPA administration markedly improved DAI,reduced colon shortening,and decreased the production of myeloperoxidase(MPO),malondialdehyde(MDA),ROS,IL-1β,IL-18,and NOD-like receptor protein 3(NLRP3)inflammasome-associated genes/proteins in UC mice.CP-A treatment also elevated glutathione(GSH)and superoxide dismutase(SOD)levels,stimulated autophagy(LC3B,P62,Beclin-1,and ATG5),and reinforced Claudin-1 and ZO-1 expression,thereby aiding in intestinal epithelial barrier repair in colitis mice.Notably,the inhibition of autophagy via chloroquine(CQ)diminished CP-A’s protective impact against colitis in vivo.These findings elucidate that CP-A’s therapeutic effect on experimental colitis possibly involves mitigating intestinal inflammation through autophagymediated NLRP3 inflammasome inactivation.Consequently,inulin-type fructan CP-A emerges as a promising drug candidate for UC treatment.
基金National Natural Science Foundation of China(Grant No.21977005)the Beijing Natural Science Foundation(Grant No.7192101)。
文摘To develop an HPLC-ELSD method for the determination of nine inulin-type fructo-oligosaccharides in inulin,the HPLC-ELSD system consisted of Waters XBridge■ Amide column(4.6 mm×250 mm,5μm)with a gradient elution mobile phase consisting of acetonitrile and water at a flow rate of 1.2 mL/min at 30°C.The detector was an Agilent Technologies 380-ELSD.The drift tube temperature for the ELSD was set at 55°C with a nitrogen flow rate of 1.8 L/min.The injection volume was 15μL.The results showed that the detection range for the nine inulin-type fructo-oligosaccharides was 3.81–30.60μg R^(2)=0.99969 for kestose,3.73–29.97μg R^(2)=0.99981 for nystose,3.82–30.69μg R^(2)=0.99993 for fructosylnystose,3.80–30.48μg R^(2)=0.99995 for GF5,3.73–29.96μg R^(2)=0.99993 for GF6,3.78–30.30μg R^(2)=0.99983 for GF7,3.82–30μg R^(2)=0.99989 for GF8,3.71–29.80μg R^(2)=0.99974 for GF9,3.61–29.00μg R^(2)=0.99970 for GF10,respectively.The recovery of the nine oligosaccharides ranged between 96.48%–100.84%(n=6).The method was simple,accurate,and reproducible that it could be used as an analytical method for evaluating the quality of inulin effectively.
基金funded by the National High-Tech R&D Program of China (863 Program of China,2007AA10Z129)
文摘Fructan is not only a carbon source for storage but also plays an important role as anti-stress agents in many plant species. Complex fructans having both β-（2,1）- and β-（2,6）-linked fructosyl units accumulate in Triticeae plants commonly. Three enzymes （sucrose： sucrose 1-fructosyltransferase, 1-SST, EC： 2.4.1.99; sucrose： fructan 6-fructosyltransferase, 6- SFT, EC： 2.4.1.10; and fructan： fructan 1-fructosyltransferase, 1-FFT, EC： 2.4.1.100） were involved in fructan biosynthesis in Triticeae plant species. We successfully isolated these genes from tetraploid wheat （Triticum turgidum, genotype： AABB）, common wheat （Triticum aestivum L., genotype： AABBDD） and three wild relatives of common wheat, Triticum urartu Thum. （the origin of the AA genome）, Aegilops speltoides （Tausch） Gren. （the putative source of the SS genome） and Aegilops tauschii Coss. （the source of the DD genome）. Sequence analysis revealed that all the FBEs （fructan biosynthetic enzymes） had three highly conserved functional motifs except 1-SST （EU981912） from tetraploid wheat species only with conserved DPNG. Low pI （isoelectric point） and potential N-glycosylation sites were predicted, which were crucial for protein compartmentation and post-translational process. Analysis on subcelluar localization signals showed that only 6-SFT had vacuolar-directed signal. Sequences alignment result showed that 1-SST and 1-FFT were more conservative and had closer relationship each other, while 6-SFT was more active during the evolution processing. According to the syntenic relationship between wheat and rice genome, FBEs were predicated to be located on the homeologous group 6 and group 2 chromosomes. Expression profile confirmed that expression of all the three FBEs were drought-stress induced. This study can assist to establish a useful theoretical platform for cold- or drought-tolerant improvement of wheat by modulating FBEs expression.
文摘Translocation of previously accumulated nitrogen and carbohydrates from vegetative tissue of the wheat plant is a major assimilate source for grain filling. This study was conducted to examine genotype differences in nitrogen and fructan translocation and their relationships to grain yield and protein content. Effects indicated that significant genotype differences existed for nitrogen accumulation at anthesis and fructan at milk stage and their translocation. Two high protein genotypes, Cunningham and PST90-19, accumulated more nitrogen before anthesis and had greater nitrogen translocation, but lower post-anthesis nitrogen uptake, than two low protein genotypes, SUN109A and TM56. Among plant parts, leaves were the major storage for tissue nitrogen and provided the overwhelming proportion of the total nitrogen translocation, whereas for fructan accumulation and translocation it was the stems. The two high protein genotypes had a higher percentage of their grain nitrogen derived from nitrogen translocation, while for the two low protein ones, it was from post-anthesis nitrogen uptake and assimilation. Increasing nitrogen application increased nitrogen accumulation and translocation, but decreased fructan accumulation and translocation. High grain protein content was associated with high nitrogen translocation from leaves, stems and the total plant, while high grain yield was related to high fructan translocation from stems and the total plant. Fructan translocation was negatively correlated to grain protein content. Nitrogen and fructan translocation were not correlated with each other.
文摘The aim of the study was to contribute to the information on agave soluble fibers since research has been focused on chicory fiber but not in agave products;thus we assess the acute toxicity and genotoxicity of two organic and high purity dietary soluble fibers from agave, Metlin? and Metlos?. We performed an acute toxicity assay in Hsd:ICR mice and Hsd:Wistar rats and an in vivo genotoxic test. Results showed that there are no deaths at any doses or genotoxicity, so it can be concluded that these products are non-toxic, at the administrated doses and none showed a cytotoxic, clastogenic or aneuploidic effect.
基金Financial support from BarleyFunFood–Barley for Functional Food and Feed,a joint program granted by the Faculty of Natural Resources and Agricultural Sciences,SLU,and Lantmännen AB is gratefully acknowledged.
文摘Fructans are important in the survival of plants and also valuable for humans as potentially health promoting food ingredients. In this study fructan content and composition were determined in grains of 20 barley breeding lines and cultivars with a wide variation in chemical composition, morphology and country of origin, grown at one site in Chile. There was significant genotypic variation in grain fructan content ranging from 0.9% to 4.2% of grain dry weight. Fructan degree of polymerisation (DP) was analysed using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Changes in the distribution of different chain lengths and the pattern of structures of fructan were detected with increasing amount of fructan in the different barleys. A positive correlation was found between fructan content and the relative amount of long chain fructan (DP > 9) (r = 0.54, p = 0.021). Our results provide a basis for selecting promising barley lines and cultivars for further research on fructan in barley breeding with the aim to produce healthy food products.
文摘为探究乳酸菌果聚糖代谢机制和开发低果聚糖发酵食品,本研究对287株酸面团来源的乳酸菌进行果聚糖利用能力筛选,获得1株嗜果聚糖乳酸菌19M03,之后利用高通量测序和生物信息学技术对该菌株进行全基因组测序和果聚糖代谢机制分析。测序结果表明,菌株19M03基因组含有1个染色体和9个质粒,基因组全长3246316 bp,GC相对含量44.53%,包含3084个编码基因、66个tRNA和16个rRNA。系统发育关系分析表明,分类学上菌株19M03属于植物乳植杆菌。基因功能注释结果表明,在同源蛋白簇、基因本体论、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)和碳水化合物活性酶数据库中,19M03共编码357个碳水化合物代谢相关基因和351个碳水化合物转运相关基因,其中与果聚糖代谢和转运相关的基因均有12个。根据这些基因在KEGG数据库中的代谢关系,提出19M03果聚糖代谢模型。该模型解析了果聚糖水解过程及其催化反应的细胞定位(既可胞内,又可胞外)、细胞膜中运输低聚果糖和果糖的特异转运蛋白(IIABC^(FOS)、IIABCD^(fru)、IIABC^(fru))、细胞内果糖分解代谢途径(n=4)和果糖合成代谢途径(n=1)及其所有相关酶的编码基因。此外,耐药和毒力基因分析证明19M03具有良好的安全性。总之,本研究筛选获得了1株嗜果聚糖乳酸菌19M03,并在全基因组水平揭示了19M03果聚糖代谢机制,这些结果可为阐明乳酸菌果聚糖代谢机制提供重要的理论参考,为今后低果聚糖功能食品的开发和应用提供优质的菌种资源。
