[Objective] To analyze the components and structure of Opuntia milpa alta polysaccharides, and to study its anticoagulant activity. [Method] The crude polysaccharides （CP） were prepared from Opuntia milpa alta using...[Objective] To analyze the components and structure of Opuntia milpa alta polysaccharides, and to study its anticoagulant activity. [Method] The crude polysaccharides （CP） were prepared from Opuntia milpa alta using hot water extraction and ethanol precipitation method, purified by anion-exchange chromatography and gel filtration chromatography to study its monosaccharide composition, weight-average molecular weight distribution and main chain structure. And a preliminary study was conducted on the anticoagulant activity of polysaccharide components. [Result] The polysaccharide components WSP1, WSP2a and WSP3 were obtained from CP by the anion-exchange chromatography and gel filtration chromatography. WSP1 was mainly composed of galactose with a weight-average molecular weight of 2.32×106. WSP2a consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid, the contents were 6.3%, 32.3%, 12.9%, 1.5%, 4.8%, 37.1%, 0.64% and 4.4%, respectively. WSP3 mainly composed of arabinose, xylose, rhamnose and trace of glucose, mannose, uronic acid. The weight-average molecular weight of WSP2a and WSP3 was 1.24×106 and 7.92×106, respectively. WSP2 prolonged the activated partial thromboplatin time （APTT） and thrombin time （TT）, while had no significant effect on the prothrombin time （PT） in vitro, indicating that anticoagulant activity of WSP2 functions by affecting the intrinsic coagulation system. The test of inhibition to the conversion of fibrinogen into thrombin showed that WSP2a and WSP3 had obvious anticoagulant activity. Methylation analysis indicated that the main chain of WSP2a was constituted of 1, 4-GalA, 1, 2-Rha and 1, 2, 4-Rha. [Conclusion] This study provided experimental basis for the optimization and development of polysaccharide resources from cactus展开更多
More and more studies have reported the role of Paecilomyces hepiali polysaccharides in bidirectional regulation(immune enhancement and anti-inflammatory).However,there are few collaborative studies on the structure,i...More and more studies have reported the role of Paecilomyces hepiali polysaccharides in bidirectional regulation(immune enhancement and anti-inflammatory).However,there are few collaborative studies on the structure,immune enhancement,and anti-inflammatory effects of polysaccharides from the fermentation broth of P.hepiali(PHP)and the mechanisms are still unclear.The present study aimed to clarify the mechanism underlying the dual-immunomodulatory(immune enhancement and anti-inflammatory).PHP40 was precipitated with 40%ethanol from PHP40 and further purified to obtain PHP40-2.The chemical characteristics of PHP40-2 was analyzed by chemical and instrumental analyses,and the double immunomodulatory effects of RAW264.7 macrophages were investigated.PHP40-2 was a heteropolysaccharide composed of→4,6)-α-D-Glcp-(1→,→6)-α-D-Manp-(1→,α-D-Glcp-(1→,and→4)-β-D-Galp-(1→,and its molecular weight was 6.2×10^(5) Da.The nitric oxide(No)production,phagocytosis activity,and cytokine secretion were significantly increased(P<0.05)after PHP40-2 treatment;however,PHP40-2 could inhibit No production and cytokine secretion in LPS-induced RAW264.7 macrophages(P<0.05).After mRNA and protein expression analysis,toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway was discovered to play an important role in the dual-immunomodulatory effects of PHP40-2.This study reveals that the fermentation broth of PHP is also a valuable resource and PHP40-2 has the potential to be developed as a novel functional food.展开更多
The potent anti-cancer metastatic effects induced by the immunostimulatory activities of intravenously administered BF–I,an active polysaccharide purified from fermented barley,were previously evaluated.This study ai...The potent anti-cancer metastatic effects induced by the immunostimulatory activities of intravenously administered BF–I,an active polysaccharide purified from fermented barley,were previously evaluated.This study aimed to evaluate the anti-cancer metastatic effect of orally administered BF–I and elucidated the microstructure and active moiety of BF–I using sequential enzymatic hydrolysis.In experiments using Colon26-M3.1 carcinoma,prophylactic oral administration of BF–I potently suppressed lung cancer metastasis.Furthermore,gas chromatography-mass spectrometry analysis of partially methylated alditol acetate showed that BF–I comprised 15 different glycosyl linkages,including 4-linked Xylp(16.1%),terminal Araf(12.4%),4-linked Glcp(32.8%),3-linked Glcp(13.5%),3,6-linked Glcp(1.5%),and 3,6-linked Galp(5.3%).These glycosyl linkage combinations are typical for arabinoxylan,yeast-derivedβ-glucan,barley-derivedβ-glucan,and type II arabinogalactan.To clarify the microstructure,BF–I was sequentially degraded via linkage-specific enzymatic hydrolysis using five enzymes.Furthermore,the sugar composition,glycosyl linkage,and reactivity with theβ-glucosyl Yariv reagent were analyzed,along with sequencing based on tandem mass spectra of partially degraded fragment fractions.The results confirmed that BF–I comprises a mixture of four different types of polysaccharides(arabinoxylan,yeast-derivedβ-glucan,barley-derivedβ-glucan,and type II arabinogalactan)with similar molecular weights.Furthermore,different polysaccharides contribute significantly to cytokine(interleukin-6,-12,and tumor necrosis factor-α)secretion by peritoneal macrophages.Therefore,the potent anti-cancer metastatic activity of BF–I is proposed to be related to the immunostimulatory activities of the four different types of polysaccharides.展开更多
Compression coated tablets for oral colon specific delivery systems were developed with a mixture polysaccharide of konjac glucomannan(KGM)and xanthan gum(XG)as the compression coat.Diffusion of cimetidine from compre...Compression coated tablets for oral colon specific delivery systems were developed with a mixture polysaccharide of konjac glucomannan(KGM)and xanthan gum(XG)as the compression coat.Diffusion of cimetidine from compression coated tablets was investigated by release experiment in Vitro.0.22U/mLβ-mannanase was applied in the mimic colon solution.The structure of the mixture polysaccharide was studied by an atomic force microscope(AFM).The experimental results indicate that a KGM70 tablet with a 0.4 g coat is of good design,due to a less than 5%drug loss in the mimic upper gastrointestinal solution by the synergistic interaction between XG and KGM,and due to about 50%cumulative release in the mimic colon solution by degradation after 24 hours.The release mechanism and model are discussed based on different periods of drug release including the delay of the drug,the constant release without an enzyme and the delay of degradation.Under hydrolysis byβ-mannanase,drug release from the tablet with KGM coat shows an exponential increase,while that from the dosage with the mixture polysaccharide coat is an approximately zero-order process in which the constant release rate relates to the release velocity of a non-degraded system,the content of KGM within the coat and the average molecular weight ratio of KGM to XG.It was found that XG was the framework of the polysaccharide mixtures by AFM,which is similar to the analysis results from experiments on drug release.展开更多
A polysaccharide of Thea sinensis, TSA, has been isolated from the fresh leaves as a major fraction of polysaccharides. The mol. wt. was estimated to be 850,000 with [α]_D^(15)+ 25.5°(c=0.75, H_2O). The componen...A polysaccharide of Thea sinensis, TSA, has been isolated from the fresh leaves as a major fraction of polysaccharides. The mol. wt. was estimated to be 850,000 with [α]_D^(15)+ 25.5°(c=0.75, H_2O). The component sugars were determined as L-rhamnose, L-arabinose, and D-galactose in molar ratio of 0.54: 1.0: 0.94. The O-acetyl groups locating on galactose residues position 2 were also iden- tified and the content was 3.8％. The ^(13)C NMR spectrum and CrO_3 oxidation of TSA indicated the Rhamnose, Arabinose and Galactose to be in α-, α- and β-configurations, respectively. Me- thylation analysis, periodate oxidation, partial hydrolysis and ^(13) C NMR. spectrum showed that TSA. is a branched galactoarabinan.展开更多
基金Supported by the National Natural Science Foundation of China(31173753)the International Scientific and Technological Cooperation Project of Anhui Province(10080703035)~~
文摘[Objective] To analyze the components and structure of Opuntia milpa alta polysaccharides, and to study its anticoagulant activity. [Method] The crude polysaccharides （CP） were prepared from Opuntia milpa alta using hot water extraction and ethanol precipitation method, purified by anion-exchange chromatography and gel filtration chromatography to study its monosaccharide composition, weight-average molecular weight distribution and main chain structure. And a preliminary study was conducted on the anticoagulant activity of polysaccharide components. [Result] The polysaccharide components WSP1, WSP2a and WSP3 were obtained from CP by the anion-exchange chromatography and gel filtration chromatography. WSP1 was mainly composed of galactose with a weight-average molecular weight of 2.32×106. WSP2a consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid, the contents were 6.3%, 32.3%, 12.9%, 1.5%, 4.8%, 37.1%, 0.64% and 4.4%, respectively. WSP3 mainly composed of arabinose, xylose, rhamnose and trace of glucose, mannose, uronic acid. The weight-average molecular weight of WSP2a and WSP3 was 1.24×106 and 7.92×106, respectively. WSP2 prolonged the activated partial thromboplatin time （APTT） and thrombin time （TT）, while had no significant effect on the prothrombin time （PT） in vitro, indicating that anticoagulant activity of WSP2 functions by affecting the intrinsic coagulation system. The test of inhibition to the conversion of fibrinogen into thrombin showed that WSP2a and WSP3 had obvious anticoagulant activity. Methylation analysis indicated that the main chain of WSP2a was constituted of 1, 4-GalA, 1, 2-Rha and 1, 2, 4-Rha. [Conclusion] This study provided experimental basis for the optimization and development of polysaccharide resources from cactus
基金sponsored by Zhejiang Provincial Key Researchand Development Program(2022C04036,2023C02040)ChinaPostdoctoral Science Foundation funded project(2022M712846).
文摘More and more studies have reported the role of Paecilomyces hepiali polysaccharides in bidirectional regulation(immune enhancement and anti-inflammatory).However,there are few collaborative studies on the structure,immune enhancement,and anti-inflammatory effects of polysaccharides from the fermentation broth of P.hepiali(PHP)and the mechanisms are still unclear.The present study aimed to clarify the mechanism underlying the dual-immunomodulatory(immune enhancement and anti-inflammatory).PHP40 was precipitated with 40%ethanol from PHP40 and further purified to obtain PHP40-2.The chemical characteristics of PHP40-2 was analyzed by chemical and instrumental analyses,and the double immunomodulatory effects of RAW264.7 macrophages were investigated.PHP40-2 was a heteropolysaccharide composed of→4,6)-α-D-Glcp-(1→,→6)-α-D-Manp-(1→,α-D-Glcp-(1→,and→4)-β-D-Galp-(1→,and its molecular weight was 6.2×10^(5) Da.The nitric oxide(No)production,phagocytosis activity,and cytokine secretion were significantly increased(P<0.05)after PHP40-2 treatment;however,PHP40-2 could inhibit No production and cytokine secretion in LPS-induced RAW264.7 macrophages(P<0.05).After mRNA and protein expression analysis,toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway was discovered to play an important role in the dual-immunomodulatory effects of PHP40-2.This study reveals that the fermentation broth of PHP is also a valuable resource and PHP40-2 has the potential to be developed as a novel functional food.
基金supported by the Basic Science Research Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education(NRF-2020R1F1A1070996).
文摘The potent anti-cancer metastatic effects induced by the immunostimulatory activities of intravenously administered BF–I,an active polysaccharide purified from fermented barley,were previously evaluated.This study aimed to evaluate the anti-cancer metastatic effect of orally administered BF–I and elucidated the microstructure and active moiety of BF–I using sequential enzymatic hydrolysis.In experiments using Colon26-M3.1 carcinoma,prophylactic oral administration of BF–I potently suppressed lung cancer metastasis.Furthermore,gas chromatography-mass spectrometry analysis of partially methylated alditol acetate showed that BF–I comprised 15 different glycosyl linkages,including 4-linked Xylp(16.1%),terminal Araf(12.4%),4-linked Glcp(32.8%),3-linked Glcp(13.5%),3,6-linked Glcp(1.5%),and 3,6-linked Galp(5.3%).These glycosyl linkage combinations are typical for arabinoxylan,yeast-derivedβ-glucan,barley-derivedβ-glucan,and type II arabinogalactan.To clarify the microstructure,BF–I was sequentially degraded via linkage-specific enzymatic hydrolysis using five enzymes.Furthermore,the sugar composition,glycosyl linkage,and reactivity with theβ-glucosyl Yariv reagent were analyzed,along with sequencing based on tandem mass spectra of partially degraded fragment fractions.The results confirmed that BF–I comprises a mixture of four different types of polysaccharides(arabinoxylan,yeast-derivedβ-glucan,barley-derivedβ-glucan,and type II arabinogalactan)with similar molecular weights.Furthermore,different polysaccharides contribute significantly to cytokine(interleukin-6,-12,and tumor necrosis factor-α)secretion by peritoneal macrophages.Therefore,the potent anti-cancer metastatic activity of BF–I is proposed to be related to the immunostimulatory activities of the four different types of polysaccharides.
基金the financial support from Grand project of Tianjin City,China(No.07ZCZDGX19600).
文摘Compression coated tablets for oral colon specific delivery systems were developed with a mixture polysaccharide of konjac glucomannan(KGM)and xanthan gum(XG)as the compression coat.Diffusion of cimetidine from compression coated tablets was investigated by release experiment in Vitro.0.22U/mLβ-mannanase was applied in the mimic colon solution.The structure of the mixture polysaccharide was studied by an atomic force microscope(AFM).The experimental results indicate that a KGM70 tablet with a 0.4 g coat is of good design,due to a less than 5%drug loss in the mimic upper gastrointestinal solution by the synergistic interaction between XG and KGM,and due to about 50%cumulative release in the mimic colon solution by degradation after 24 hours.The release mechanism and model are discussed based on different periods of drug release including the delay of the drug,the constant release without an enzyme and the delay of degradation.Under hydrolysis byβ-mannanase,drug release from the tablet with KGM coat shows an exponential increase,while that from the dosage with the mixture polysaccharide coat is an approximately zero-order process in which the constant release rate relates to the release velocity of a non-degraded system,the content of KGM within the coat and the average molecular weight ratio of KGM to XG.It was found that XG was the framework of the polysaccharide mixtures by AFM,which is similar to the analysis results from experiments on drug release.
文摘A polysaccharide of Thea sinensis, TSA, has been isolated from the fresh leaves as a major fraction of polysaccharides. The mol. wt. was estimated to be 850,000 with [α]_D^(15)+ 25.5°(c=0.75, H_2O). The component sugars were determined as L-rhamnose, L-arabinose, and D-galactose in molar ratio of 0.54: 1.0: 0.94. The O-acetyl groups locating on galactose residues position 2 were also iden- tified and the content was 3.8％. The ^(13)C NMR spectrum and CrO_3 oxidation of TSA indicated the Rhamnose, Arabinose and Galactose to be in α-, α- and β-configurations, respectively. Me- thylation analysis, periodate oxidation, partial hydrolysis and ^(13) C NMR. spectrum showed that TSA. is a branched galactoarabinan.
