Sulfated Cyclocarya paliurus polysaccharides exert immunomodulatory potential on macrophages via Toll-like receptor 4 mediated MAPK/NF-κB signaling pathways

The biological activity of plant polysaccharides can be enhanced by sulfated modification.In this study,the immunomodulatory effect of sulfated Cyclocarya paliurus polysaccharides(SCP3)on macrophages RAW264.7 and its potential molecular mechanism were investigated.Results showed that SCP3 at 25-100μg/m L increased viability and improved phagocytosis of RAW264.7 cells.Meanwhile,SCP3 could activate mitogen-activated protein kinase(MAPK)and nuclear factor kappa B(NF-κB)signaling pathways,which increased the phosphorylation of Erk1/2,JNK,p38 and NF-κB p65,promoting secretion of cytokines tumor necrosis factorα(TNF-α),interleukin 6(IL-6)and nitric oxide(NO)as well as the production of reactive oxygen species(ROS).In addition,Toll-like receptor 4(TLR4)receptor inhibitors were able to block the production of NO and TNF-αby SCP3-stimulated macrophages.Based on Western blot analysis and validation using specific inhibitors against MAPK and NF-κB signaling pathways,the results demonstrated that SCP3 induced macrophages activation and enhanced TNF-αand NO production via TLR4-mediated MAPK and NF-κB pathways.In summary,SCP3 has significant immunomodulatory potential.The underlying molecular mechanism was that SCP3 activates macrophages via TLR4 receptors to promote ROS production,which in turn activates the downstream MAPK/NF-κB signaling pathway and then increases the secretion levels of cytokines and NO.

Sulfated polysaccharides with antioxidant and anticoagulant activity from the sea cucumber Holothuria fuscogliva

Sea cucumber is a traditional nutritional food and medicinal resource with many bioactive components in China.Holothuria fuscogliva is a big sea cucumber with a rich of bioactive polysaccharides.To investigate the bioactivities of the polysaccharides from sea cucumber H.fuscogliva,we prepared the sulfated polysaccharides(HfP) from sea cucumber H.fuscogliva using a protease hydrolysis method.Antioxidant activities of HfP were investigated,including hydroxyl radical scavenging activity and superoxide radical scavenging activity.And,the anticoagulant activities of HfP were studied,including the activated partial thromboplastin time(APTT),prothrombin time(PT) and thrombin time(TT).The average molecular weight was 1 867.1 Da,with a sulfate content of 20.7%.In addition,the molar ratio of monosaccharide composition of HfP was Man:Rha:Glc A:Glc:Gal:Xyl:Fuc=0.083 6:0.437:0.134:0:1.182:0.748:1.It had a strong antioxidant activity,the hydroxyl and superoxide radical scavenging activity EC_(50) of HfP was 3.74 and 0.037 mg/mL,respectively.It also showed a good anticoagulant activity in our study.The APTT of HfP was much higher than that of heparin sodium,and the PT and TT of HfP was close to that of heparin sodium at a low concentration.Therefore,HfP shows a good antioxidant and anticoagulant activity and it may become a potential candidate of the natural antioxidant and anticoagulant and will have a good application future in health product or medicine industry.

Chemical Characteristics and Anticoagulant Activities of Two Sulfated Polysaccharides from Enteromorpha linza (Chlorophyta)

Two sulfated polysaccharides, designated MP and SP, were extracted from the marine green alga Enteromorpha linza using hot water and then purified using ion-exchange and size-exclusion chromatography. The anticoagulant activities of MP and SP were examined by determination of their activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) using human plasma. Results showed that MP and SP were composed of abundant rhamnose with small amounts of xylose and glucuronic acid, whereas SP also contained a small amount of galactose. Approximate molecular weights of MP and SP were 535 and 502 kDa, respectively. As compared with SP, MP had higher contents of sulfate ester (19.0%) and uronic acid (14.9%). The MP mainly consisted of (1→4)-linked rhamnose residues with partially sulfated groups at the C-3 position, and small amounts of (1→3, 4)-linked rhamnose, (1→2, 4)-linked rhamnose, (1→4)-linked glucuronic acid and (1→4)-linked xylose residues. The SP contained abundant (1→4)-linked rhamnose with minor amounts of (1→3)-linked rhamnose, (1→3, 4)-linked rhamnose, (1→2, 4)-linked rhamnose, (1→4)-linked glucuronic acid, (1→4)-linked xylose, and (1→3)-linked galactose residues. The sulfate groups were mainly located at C-3 of (1→4)-linked rhamnose residues. Both MP and SP, in particular the former, effectively prolonged APTT and TT. This work demonstrates that MP and SP have unique structural characteristics distinct from those of other sulfated polysaccharides from Enteromorpha. The MP is a potential source of anticoagulant, and the difference in anticoagulant activities of the two sulfated polysaccharides is directly linked to the discrepancy of their chemical features.

In Vitro Anti-influenza Virus Activities of Sulfated Polysaccharide Fractions from Gracilaria lemaneiformis

In this paper, in vitro anti-influenza virus activities of sulfated polysaccharide fractions from Gracilaria lemaneiformis were investigated. Cytotoxicities and antiviral activities of Gracilaria lemaneiformis polysaccharides (PGL), Gracilaria lemaneiformis polysaccharide fraction-1 (GL-1), Gracilaria lemaneiformis polysaccharide fraction-2 (GL-2) and Gracilaria lemaneiformis polysaccharide fraction-3 (GL-3) were studied by the Methyl thiazolyl tetrazolium (MTT) method, and the inhibitory effect against Human influenza virus H1-364 induced cytopathic effect (CPE) on MDCK cells were observed by the CPE method. In addition, the antiviral mechanism of PGL was explored by Plaque forming unit (PFU), MTT and CPE methods. The results showed: i) Cytotoxicities were not significantly revealed, and H1-364 induced CPE was also reduced treated with sulfated polysaccharide fractions from Gracilaria lemaneiformis; ii) Antiviral activities were associated with the mass percentage content of sulfate groups in polysaccharide fractions, which was about 13%, in polysaccharides (PGL and GL-2) both of which exhibited higher antiviral activity; iii) A potential antiviral mechanism to explain these observations is that viral adsorption and replication on host cells were inhibited by sulfated polysaccharides from Gracilaria lemaneiformis. In conclusion, Anti-influenza virus activities of sulfated polysaccharide fractions from Gracilaria lemaneiformis were revealed, and the antiviral activities were associated with content of sulfate groups in polysaccharide fractions.

Inhibition of the Crystal Growth and Aggregation of Calcium Oxalate by Algae Sulfated Polysaccharide In-vitro

The influence of sulfated polysaccharide （SPS） isolated from marine algae Sargassum fusiforme on the morphology and phase compositions of urinary crystal calcium oxalate was investigated in vitro by means of scanning electron microscopy and X-ray diffraction. SPS maybe is a potential inhibitor to CaOxa urinary stones by inhibiting the growth of calcium oxalate monohydrate （COM）, preventing the aggregation of COM, and inducing the formation of calcium oxalate dihydrate （COD） crystals.

Microanalysis and preliminary pharmacokinetic studies of a sulfated polysaccharide from Laminariajaponica

A rapid,sensitive and reproducible high performance liquid chromatography(HPLC) method with post-column fluorescence derivatization has been developed to determine the amount of low-molecularweight sulfated polysaccharide(GFS) in vivo. The metabolism of GFS has been shown to fit a two component model following its administration by intravenous injection,and its pharmacokinetic parameters were determined to be as follows: half-time of distribution phase(t_(1/2α))=11.24±2.93 min,half-time of elimination phase(t_(1/2β))=98.20±25.78 min,maximum concentration(C_(max))=110.53 μg/mL and peak time(T_(max))=5 min. The pharmacokinetic behavior of GFS was also investigated following intragastric administration. However,the concentration of GFS found in serum was too low for detection,and GFS could only be detected for up to 2 h after intragastric administration(200 mg/kg body weight). Thus,the bioavailability of GFS was low following intragastric administration because of the metabolism of GFS. In conclusion,HPLC with postcolumn derivatization could be used for quantitative microanalysis and pharmacokinetic studies to determine the presence of polysaccharides in the serum following intravenous injection.

Biomechanics mechanism of the anti-tumor effects of sulfated polysaccharide and sulfated glycopeptide

There are also a variety of cytokines in the tumor microenvironment,which are involved in the change in the hardness of the tumor,thereby affecting the invasion and metastasis of tumor cells.As traditional Chinese medicines,drugs of softening and dissipating firm knot contains different kinds of sulfated polysaccharide and sulfated glycopeptide.By inhibiting the function of cancer-associated fibroblasts,they reduce interstitial fibrosis,thereby reducing the hardness of the tumor and exerting an anti-tumor effect.

Suppressive effect of pectic polysaccharides extracted from Rauwolfia verticillata(Lour.) Baill.var.hainanensis Tsiang on inflammation by regulation of NF-κB pathway and interleukin-17 in mice with dextran sulphatesodium-induced ulcerative colitis

Objective:To investigate the effects of pectic polysaccharides extracted from Rauwolfia verticillata(Lour.) Baill.var.hainanensis Tsiang on an experimental murine colitis model.Methods:Experimental colitis was induced by dextran sulfate sodium(DSS),and mice were divided into 4 groups:control.DSS alone.DSS plus SASP,DSS plus pectic polysaccharides.The disease activity index(DAI) and histological score were observed.The tumor necrosis factor(TNF)-α and interleukin(IL)-17 levels were measured by enzyme-linked immunosorbent assay.I κ B and NF-κB p65 expression were assessed by western blot analysis.Myeloperoxidase(MPO) activity was determined by using MPO assay kit.Re.sults:Administration of pectic polysaccharides significantly reduced the severity of DSS-induced colitis as assessed by DAT and histological score,and resulted in down regulation of MPO activity and NF-κB p65 expression and subsequent degradation of IκB protein,strikingly reduced the production of TNF-α and IL-17.Conclusions:Pectic polysaccharides extracted from Rauvolfia verticillata(Lour.)Baill.var.hainanensis Tsiang exerts beneficial effects in experimental colitis and may therefore provide a useful therapeutic approach for the treatment of UC.

Production and characterization of an extracellular polysaccharide of antarctic marine bacteria Pseudoalteromonas sp. S-15-13

Twenty-seven antarctic bacteria producing extracellular polysaccharide （EPS） were selected from 57 strains by staining technology. The effects of major environmental factors on the growth and EPS production of Pseudoalteromonas sp. S - 15 - 13 were investigated, and the EPS was separated and purified for characterization analysis. The results showed that the optimal conditions for the EPS production were culture period, 56 h; growth temperature, 8 ℃ ; carbon source, 1.0% glucose; NaCI concentration, 3.0% ; pH 6.0 - 7.0. The EPS was purified by cold ethanol precipitation, proteins removal, ion exchange chromatography and gel chromatography technology. The molecular mass of EPS - H was 62 kDa as determined by the high performance gel permeation chromatography. Its sugar composition was a homopolymer of marmose analyzed by gas chromatograph spectroscopy. After repeated freezing and thawing of the bacteria hiomass in the presence of EPS, the bacterial growth was much higher than that observed after freezing in the absence of EPS and the difference augmented with the increase of freeze-thaw cycles. It is hypothesized that the adaptation of Pseudoalteromonas sp. S- 15 - 13 to the antarctic marine conditions, characterized by low temperature, high NaCl concentration and repeated freeze-thaw cycles, might be related to the EPS production ability.

Isolation of a novel strain of Cyanobacterium sp.with good adaptation to extreme alkalinity and high polysaccharide yield

The use of high alkaline medium is a feasible way to provide carbon source and prevent biological contamination for the outdoor cultivation of alkaliphilic microalgae and cyanobacteria.A novel cyanobacterial strain was isolated from the open pond of a marine green alga(Picochlorum sp.SCSIO-45015,Sanya,Hainan)and identified as Cyanobacterium sp.SCSIO-45682.The effects of initial sodium bicarbonate(NaHCO_(3))concentrations on the growth and biochemical composition of Cyanobacterium sp.SCSIO-45682 were investigated.The results demonstrated that Cyanobacterium sp.SCSIO-45682 had good adaptation to 16.8-g/L NaHCO_(3)(the same concentration of NaHCO_(3) used in Zarrouk medium for Spirulina).Moreover,the yields of biomass,polysaccharide,chlorophyll a(chl a),and phycocyanin increased under high NaHCO_(3) concentrations.The maximum final biomass concentration of 2.5 g/L was observed at 8.4-g/L NaHCO_(3),while the highest intracellular total saccharide content of 49.2%of dry weight(DW)and exopolysaccharide(EPS)concentration of 93 mg/L were achieved at the NaHCO_(3) concentration of 16.8 g/L.The crude protein content declined under high NaHCO_(3) concentrations,which provide a possible explanation for the accumulation of polysaccharide.This study shows a good potential of alkaliphilic Cyanobacterium sp.SCSIO-45682 as a polysaccharide feedstock.

裙带菜多糖抑制肝癌细胞增殖和迁移的机制

目的探讨裙带菜多糖(sulfated polysaccharide from Undaria pinnatifida,SPUP)对肝癌细胞增殖、迁移的影响及潜在机制。方法MTT和克隆形成实验检测SPUP对肝癌细胞HepG2、SMMC-7721增殖的影响;Hoechst 33258染色和Western blot法检测SPUP干预后对HepG2、SMMC-7721细胞凋亡的影响;Transwell和划痕法检测HepG2、SMMC-7721细胞的迁移和侵袭;免疫荧光和Western blot法检测SPUP对肝癌细胞自噬、上皮-间质转化(epithelial-mesenchymal transition,EMT)和基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)、MMP-9蛋白的影响。结果SPUP对HepG2、SMMC-7721细胞有短期和长期的抑制增殖作用,对正常肝细胞L-02无明显抑制作用,SPUP可促进两种肝癌细胞凋亡和自噬水平。同时,SPUP可以抑制肝癌细胞的迁移侵袭能力,降低EMT和MMPs水平。结论SPUP通过促进凋亡和自噬、抑制EMT和MMPs,进而抑制肝癌细胞的增殖和迁移。

响应面法优化茭白多糖硫酸化工艺及其抗氧化活性分析

为提高茭白多糖抗氧化活性,对茭白多糖进行硫酸化修饰,并探索其最佳修饰条件。以茭白为原料,利用水提醇沉法提取并纯化一种新型水溶性多糖,采用三氧化硫-吡啶法,以甲酰胺为溶剂,对多糖进行硫酸改性,以硫酸根取代度(DS)为指标,利用响应面法对茭白多糖进行硫酸改性条件优化,同时通过体外自由基清除率实验对比改性前后体外抗氧化活性。结果表明:采用响应面法辅助茭白多糖硫酸化修饰条件优化得到最佳反应条件为多糖样品100 mg,酯化时间4 h,酯化温度70℃,三氧化硫-吡啶复合物添加量2.6 g,得到最佳DS为1.79;热重分析显示茭白多糖在46.38℃时失重最快,失重率为16.48%;硫酸化茭白多糖在65.04℃时失重最快,失重率为2.92%。在体外抗氧化实验中,茭白多糖对于DPPH·、·OH以及ABTS^(+)·清除能力的IC_(50)值分别为3.49、3.28和12.70 mg/mL;硫酸化茭白多糖为0.39、1.00和1.82 mg/mL;阳性对照(V_(C))为0.069、0.17和0.077 mg/mL。综上表明,硫酸化修饰能提高茭白多糖的热稳定性;同时一定程度上增强其自由基清除能力。

螺旋藻多糖PSP的化学结构研究

目的:研究螺旋藻多糖硫酸脂的理化性质及化学结构。方法:采用高效液相色谱、气相色谱、特异性酶降解、高碘酸氧化、Smith降解、部分酸水解测定PSP的分子量、单糖组成、连接方式、糖苷键键型、主链和支链结构及分枝点。结果:螺旋藻多糖是由多种单糖组成的杂多糖,具有β型糖苷键,主链部分以1→3键连接为主。结论:螺旋藻多糖是一种结构复杂的新型多糖。

硫酸—蒽酮分光光度法测定SPS的方法研究

对硫酸—蒽酮分光光度法测定硫酸酯化多糖 (SPS)含量方法进行了考察 .实验结果表明 ,水解反应—显色时间 1 0～ 1 5min ,蒽酮—硫酸试剂加入量 3.0～ 4.0ml,称样量 2 0～ 30mg ,均能得到满意的结果 .所测结果可靠 ,重复性好 ,回收率为 98.5 % .该法可用于硫酸酯化多糖和相关食品、保健食品中多糖的常规质量控制分析 .

温度对铜绿微囊藻(Microcystis aeruginosa)和鱼腥藻(Anabaena sp.)生长及胞外有机物产生的影响

以巢湖优势种铜绿微囊藻(Microcystis aeruginosa)和鱼腥藻(Anabaena sp.)为研究对象,研究不同温度(35、25和10℃)对这两种藻生长特性和胞外有机物产生的影响.结果表明,温度对铜绿微囊藻和鱼腥藻的藻细胞密度、碱性磷酸酶活性和胞外有机物浓度影响显著.25℃是铜绿微囊藻和鱼腥藻最适宜的生长温度,最高细胞密度分别达到3.12×107cells/ml和2.03×107cells/ml.不同温度下两种藻的碱性磷酸酶活性特征,证实了高温对鱼腥藻生长的抑制和低温对铜绿微囊藻生长的抑制.胞外有机物释放总量受蓝藻生物量和单位细胞有机物释放速率的影响.铜绿微囊藻的溶解性有机碳和胞外总多糖释放量在25℃最高,最大值分别为49.28和38.46 mg/L;而鱼腥藻在35℃时释放量最高,最大值分别为45.82和40.60 mg/L;10℃条件抑制了两种藻的生长及胞外有机物的释放.鱼腥藻胞外多糖含量在35℃培养条件下最高,而铜绿微囊藻在10℃条件下最高,说明不利的生长条件会促进蓝藻胞外多糖的分泌.三维荧光图谱分析结果表明,铜绿微囊藻和鱼腥藻胞外有机物以类蛋白质和类腐殖酸为主,温度主要影响藻细胞胞外有机物浓度,而对有机物种类组成没有影响.

药用真菌桑黄(phellinus sp．)抗癌功能的研究进展

桑黄(phellinus sp．)是一种近年来由韩国、日本兴起开发的稀有药用真菌,有着很好的抗癌效果。本文主要从提高机体免疫、抑制癌转移、调节药物代谢酶等多个角度,着重介绍桑黄多糖和小分子天然产物的抗癌功效和机理,并提出了以后桑黄抗癌药物深入开发的研究方向。

响应面法优化Chlamydomonassp.212产胞内多糖发酵工艺及其抑菌活性

通过响应面法优化淡水微藻Chlamydomonas sp.212产胞内多糖的发酵工艺,并采用滤纸片法对其抑菌活性进行研究。经优化后,Chlamydomonas sp.212产胞内多糖发酵工艺的最优参数为Na NO3质量浓度305.01 mg/L、NaCl质量浓度93.66 mg/L、NaHCO_3质量浓度2.12 g/L;在此条件下,其胞内多糖产量为91.182 3 mg/L,比优化前提高了1.6倍。抑菌活性研究结果表明:Chlamydomonas sp.212的胞内多糖对细菌有一定程度的抑制作用,其中对沙门氏菌的抑菌作用最强,对大肠杆菌、白色葡萄球菌、金黄色葡萄球菌的抑制作用较弱;对真菌的抑制作用较弱,仅对黑曲霉有较弱的抑制作用。本研究为筛选新的天然抗菌活性物质及产油微藻的综合开发利用提供了一定的理论依据。

南极适冷菌Pseudoalteromonas sp. S-15-13胞外多糖的分离、纯化和结构分析

从一株南极海冰中分离出来一种适冷菌Pseudoalteromonas sp. S-15-13,其胞外多糖具有良好的免疫活性.为了探讨南极菌S-15-13胞外多糖结构与功能之间的相互关系,对其胞外多糖进行了分离纯化和结构分析.粗多糖经DEAE-Sephadex A-50离子交换层析及Sephadex G-100凝胶层析纯化后得到组分EPS-Ⅱ,经HPLC分析验证EPS-Ⅱ为单一组分,其分子量为62000;单糖组成、甲基化分析及核磁共振结果表明,EPS-Ⅱ的主体结构由(1,2)α-D-Man组成主链,并在6位上有分支的新甘露聚糖.

苔海绵Tedania sp.多糖及其中硫酸根含量测定

为测定苔海绵Tedania sp.多糖及其中硫酸根含量,分别用水提和酸提法制备粗多糖,用苯酚-硫酸法显色,测定多糖含量,并用硫酸钡比浊法对其中的硫酸根含量进行了测定。测得水提和酸提粗多糖中总糖含量分别为16.79%,20.55%;硫酸根含量分别为24.02%,27.02%。结果说明酸提法得到的粗多糖含量及其中硫酸根的含量大于水提法。

营养因子对钝顶螺旋藻Sp-S(9701)生长与多糖合成的影响

研究了培养液中Mg2+、K+、NO3-和HCO3-的不同浓度对钝顶螺旋藻Sp S(9701)生长及多糖合成的影响。结果表明,培养液中Mg2,K+,NO3-和HCO3-的浓度分别为10mg/L,560mg/L,876mg/L和12.3g/L时,是Sp S(9701)高产多糖的最佳条件。