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1.
目的从扁玉螺(Neverita didyma)中提取和分离纯化多糖,并对其基本理化性质和结构组成进行分析。方法依次采用水提和碱提方法从扁玉螺中提取粗多糖,采用DEAE Sepharose FF阴离子交换和Sephacryl S-300凝胶柱层析对粗多糖进行分离纯化,并对其总糖、蛋白、氨基糖、糖醛酸和硫酸根含量,相对分子质量和单糖组成进行分析。对多糖纯化组分采用甲基化、气质联用(GC-MS)、红外光谱(IR)、电喷雾质谱(ESI-MS)和核磁共振波谱(NMR)对其化学结构进行分析。结果扁玉螺水提粗多糖(BYL-S)中不含有糖醛酸和硫酸基,其单糖组成只含Glc,进一步分离得到了4种水溶性多糖组分。碱提粗多糖(BYL-J)单糖组成相对复杂,除含有Glc外,还含有Man、GlcN、GalN、Gal和Fuc,其摩尔比为Glc∶Man∶GlcN∶GalN∶Gal∶Fuc=78.9∶1.7∶3.4∶2.2∶5.2∶5.6,进一步分离得到了3种多糖组分。对水提多糖纯化组分BYL-S2的结构分析表明其是以α-(1→4)为主链,含有少量β-(1→3,4)和β-(1→3)分支的D-吡喃型葡聚糖。结论从扁玉螺中提取分离得到了7种多糖组分,并确定了一种水溶性葡聚糖的结构,为扁玉螺多糖结构和活性的深入研究提供了基础。  相似文献   

2.
目的 从多棘海盘车(Asterias amurensis)内脏中提取、分离纯化多糖,并对其基本理化性质和结构进行表征。方法 采用沸水提取与酶解联用的方法从多棘海盘车内脏脱脂粉中提取粗多糖;采用Q Sepharose FF强阴离子交换和Sephacryl S-100凝胶渗透色谱对粗多糖进行分离纯化;采用硫酸-苯酚法、Folin-酚法、高效凝胶渗透色谱-十八角激光散射仪(HPGPC-MALLS)和高效液相色谱(HPLC)分别进行总糖含量,蛋白含量,分子量和单糖组成进行分析;采用甲基化、红外光谱(IR)、气质联用(GC/MS)和一维、二维核磁共振波谱(NMR)法对纯化多糖结构进行表征。结果 从多棘海盘车内脏中提取粗多糖的收率为3.3%。经过离子交换和分子排阻色谱分离纯化,得到一种分子量均一的多糖组分 F2-A,该多糖主要由葡萄糖(Glc)组成,其糖含量为97.6%,分子量为2708 kDa,是一种以α-(1-4)-Glc为主链并含有少量β-(1-3,4)分支的海星内脏来源葡聚糖。结论 从多棘海盘车内脏中纯化得到一种高分子量、结构独特的葡聚糖,为将来从事其结构和生物活性关系的研究提供了有用信息。  相似文献   

3.
目的从缢蛏中提取和分离纯化多糖,对其基本理化性质和结构组成进行分析。方法依次采用100℃热水提和碱提方法从缢蛏中提取粗多糖,采用Q Sepharose Fast Flow强阴离子交换柱层析和Sephacryl S-300凝胶柱层析对粗多糖进行分离纯化,并对其总糖、蛋白含量,相对分子质量和单糖组成进行分析。对多糖纯化组分采用甲基化、气质联用(GC-MS)、红外光谱(IR)和核磁共振波谱(NMR)进行化学结构解析。结果从缢蛏中经热水提粗多糖(YC-S)和碱提粗多糖(YC-J)均为葡聚糖,进一步分离纯化得到了4种水溶性多糖组分。对其中1种热水提多糖纯化组分YC-S2采用高效液相凝胶色谱法测得其相对分子质量为482kD,且色谱峰单一对称,表明其具有较高的纯度。结构分析表明,YC-S2是1种以α-(1→4)Glc为主链,含有少量的β-(1→3,4)和β-(1→4)分支的D-吡喃型葡聚糖。结论从缢蛏中提取分离得到了4种多糖组分,并初步确定了1种水溶性葡聚糖的结构,为缢蛏多糖结构和活性的深入研究提供了基础。  相似文献   

4.
目的 从太平洋牡蛎(Crassostrea gigas)肉中提取和分离纯化多糖,并对其基本理化性质和结构组成进行分析。方法 将牡蛎鲜肉制成丙酮粉,依次采用室温水,60 ℃热水和稀碱提取牡蛎粗多糖,对其总糖含量、蛋白含量和单糖组成进行分析。采用DEAE Sepharose FF阴离子交换和Sephacryl S-400凝胶柱层析对粗多糖进行分离纯化,并对获得的纯化组分采用红外光谱 (IR)、甲基化、气质联用 (GC-MS)、电喷雾质谱 (ESI-MS) 和核磁共振波谱 (NMR) 等方法进行化学结构分析。结果 从牡蛎肉中提取得到了3种牡蛎粗多糖,单糖组成都只含有葡萄糖 (Glc)。对粗多糖进一步分离,得到了5种多糖纯化组分。对水溶性多糖纯化组分 MC-11 的结构进行了分析,表明其是以 →4)-α-D-Glc-(1→ 为主链,含有 →3,4)- β-D-Glc-(1→和 →2,4)- β-D-Glc-(1→ 分支的 D-吡喃型葡聚糖,相对分子质量为1299 kDa。结论 从太平洋牡蛎肉中分离纯化得到了5种多糖组分,并采用多种方法确定了1种纯化组分 MC-11 的结构,为牡蛎多糖结构与活性关系的深入研究提供了基础。  相似文献   

5.
目的对吐鲁番葡萄树伤流液多糖进行分离纯化,并分析其单糖组成。方法采用醇沉法提取多糖,Sevage法脱蛋白,经DEAE-52纤维素柱及Sephadex G-150凝胶柱分离纯化,采用气相色谱分析单糖组成。结果用体积分数80%乙醇沉淀、Sevage法除蛋白,得葡萄树伤流液粗多糖,粗多糖经过DEAE-52纤维素柱分离得到8个多糖组分,编号分别为:组分-1,组分-2,组分-3,组分-4,组分-5,组分-6,组分-7,组分-8。其中4个多糖组分经过Sephadex G-150纯化后,通过GC分析组分-1和组分2含有鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖、半乳糖;组分-3含有鼠李糖、阿拉伯糖、甘露糖、葡萄糖、半乳糖;组分-5含有鼠李糖、甘露糖、葡萄糖、半乳糖。结论该方法可较好地分离纯化葡萄树伤流液多糖。采用GC分析多糖的单糖组成,研究结果适用于多糖中的单糖组成分析。  相似文献   

6.
紫贻贝多糖的提取、分离和基本理化性质分析   总被引:1,自引:0,他引:1  
目的从紫贻贝(Mytilus edulis Linnaeus)中提取和分离纯化多糖,并对其基本理化性质进行分析,为贻贝多糖的结构和活性研究提供依据。方法依次采用冷水,木瓜蛋白酶和胰蛋白酶联合酶解的方法提取贻贝粗多糖,并对粗多糖的总糖、蛋白、糖醛酸、糖胺聚糖和硫酸根含量进行分析。分别采用Sephacryl S-300凝胶柱层析和QSepharose 4 Fast Flow阴离子交换柱层析对粗多糖进行分离和纯化,并对纯化后的组分进行单糖组成、纯度及相对分子质量分析。结果从紫贻贝中经提取和分离共得到了8种水溶性多糖组分,其中水提组分LTS1-A单糖组成较简单,只含Glc。其余各组分单糖组成相对复杂,主要含有Man、GlcN、Gal和Fuc。采用高效凝胶渗透色谱法对各多糖组分进行分析都呈现较为均一的色谱峰,表明具有较好的纯度,各组分相对分子质量范围约为3.0~40 kD。结论采用水提和酶提方法得到了具有不同理化性质的多糖,经两步层析分离能够得到纯度较高的多糖组分,为下一步紫贻贝多糖结构和活性的深入研究提供了依据。  相似文献   

7.
目的:研究川牛膝总多糖含量及单糖组分构成。方法:利用苯酚-硫酸法测定川牛膝中总多糖含量;利用红外光谱法初步探究川牛膝多糖的官能团,再采用PMP衍生-HPLC法分析川牛膝多糖中单糖组分。结果:未分离纯化的川牛膝多糖含量为90.95%,纯化后的多糖含量为98.1%;红外光谱分析表明川牛膝多糖中含有吡喃环;PMP衍生-HPLC法分析表明川牛膝多糖主要由D-甘露糖、D(+)-无水葡萄糖、D-半乳糖、D(+)-木糖及L(-)-岩藻糖组成。结论:实验所用方法方便快捷,可以有效地分析川牛膝多糖中的单糖组分。  相似文献   

8.
目的 从南海软珊瑚Sinularia sp.中提取、分离和纯化多糖,分析其基本理化性质,并对其生物活性进行初步评价。方法 采用酸性蛋白酶从软珊瑚Sinularia sp.中提取粗多糖,利用碱性蛋白酶和732型阳离子交换树脂去除蛋白,利用Q Sepharose Fast Flow强阴离子交换柱层析对多糖进行分离纯化,并对多糖组分进行总糖含量、蛋白含量和单糖组成分析。将得到的多糖组分采用三氧化硫-吡啶法进行硫酸酯化修饰并对其进行抗病毒、抗凝血生物活性评价。结果 从软珊瑚Sinularia sp.中提取分离得到3种多糖组分,单糖组成较复杂,主要含有Ara, Gal及Rha;软珊瑚多糖硫酸酯可以明显抑制HBsAg和HBeAg的表达量;体外抗凝实验中可延长APTT和PT值。结论 从软珊瑚Sinularia sp.中得到了3种多糖组分,硫酸酯化修饰的软珊瑚多糖具有良好的抗病毒、抗凝血活性,为软珊瑚多糖的深入研究提供了基础。  相似文献   

9.
目的:对桑黄多糖中的未知糖组分进行分析鉴定。方法:从桑黄子实体中经过提取分离纯化得到水溶性多糖纯品Piw-1,经三氟乙酸完全水解后用高效阴离子交换色谱检测出含5种单糖,其中4种分别为岩藻糖、半乳糖、葡萄糖和木糖,1种在单糖标准品对照中未找到。将 Piw-1乙酰化后进一步用 GC-MS 进行分析,同时用~(13)C NMR 的 DEPT-135图谱对Piw-1中的未知单糖组分进行鉴定。结果:用离子色谱、气质联用及核磁共振的分析手段鉴定了含有3-0-甲基-半乳糖的桑黄多糖的糖组成。结论:Piw-1中含有3-0-甲基-半乳糖组分。  相似文献   

10.
目的 对1种绿藻硫酸多糖的化学组成及其结构特征进行研究。方法 通过热水提取法,从1种石莼属海藻中提取硫酸多糖,采用强阴离子交换色谱和凝胶渗透色谱对多糖进行分离纯化;采用高效凝胶渗透色谱(HPGPC)、高效液相色谱以及化学方法对多糖的纯度、分子量、单糖组成和化学成分进行分析测定;通过红外光谱和气质联用方法对多糖的结构进行表征。结果 多糖HP2S在HPGPC色谱图上呈单一对称峰,分子量为90.5 kDa,硫酸根和糖醛酸含量分别为32.1%和7.4%,HP2S主要由鼠李糖组成,含有少量葡萄糖、葡萄糖醛酸、半乳糖和木糖,HP2S糖链中鼠李糖主要存在形式为(1→2)-Rhap 、(1→3)-Rhap 和(1→2, 3)-Rhap,硫酸根位于(1→3)-Rhap 的C-2位或(1→2)-Rhap 的C-3位上。结论 水溶性多糖HP2S是1种结构新颖的主要由鼠李糖组成的硫酸化多糖。  相似文献   

11.
目的 从狭鳕 (Alaska pollack)、褐牙鲆 (Bastard halibut) 和真鲷 (Red seabream) 三种鱼鳃中提取分离糖胺聚糖并对其细微结构进行比较分析。方法 利用酶法降解并以阴离子交换色谱法分离纯化糖胺聚糖,通过单糖组成和醋酸纤维素薄膜电泳对其种类进行鉴别,采用高效液相色谱法对其相对分子量和二糖组成进行比较分析,最后利用核磁共振波谱法 (1H-NMR) 对其结构进行表征。结果 从3种鱼鳃组织中提取分离得到54种多糖,其中18种是电荷和分子量均一的糖胺聚糖,它们集中在0.8 mol?L-1 NaCl洗脱部分 (P4) 和1.0 mol?L-1 NaCl洗脱部分 (P5),占多糖总量的17.4~58.4%;P4主要为硫酸软骨素C (CSC),P5主要为硫酸软骨素A (CSA) 和杂合硫酸软骨素 (HyCS),尤其,与牛气管来源CSA比较,HyCS的4S/6S比和电荷密度均较大。结论 狭鳕、褐牙鲆和真鲷鱼鳃组织中的糖胺聚糖主要为CSA、CSC和HyCS。  相似文献   

12.
Fujiwara Y  Yamamoto C  Kaji T 《Toxicology》2000,154(1-3):9-19
To investigate the effects of lead on the formation of extracellular matrix in the vascular wall, we characterized proteoglycans synthesized by cultured vascular smooth muscle cells after exposure to the metal by biochemical techniques. Confluent cultures of bovine aortic smooth muscle cells were metabolically labeled with [(35)S]sulfate or [(35)S]methionine/cysteine in the presence of lead nitrate. The amount of glycosaminoglycans (GAGs) was evaluated by the incorporation of [(35)S]sulfate into GAGs by the cetylpyridinium chloride precipitation method. The labeled proteoglycans were characterized by DEAE-Sephacel ion exchange chromatography and Sepharose CL-2B molecular sieve chromatography. The GAG M(r) and composition were analyzed by Sepharose CL-6B chromatography, and the core protein M(r) was analyzed by SDS-polyacrylamide gel electrophoresis, before and after digestion with papain or chondroitin ABC lyase. Lead significantly decreased the [(35)S]sulfate incorporation into GAGs accumulated in the cell layer and the conditioned medium. [(35)S]Sulfate-labeled proteoglycans obtained from the cell layer and the conditioned medium were separated into three peaks on DEAE-Sephacel chromatography and only the peak with the highest charge density was decreased by lead. The highly charged peak was eluted near the void volume on Sepharose CL-2B molecular sieve chromatography and sensitive to chondroitin ABC lyase on Sepharose CL-6B chromatography, indicating that lead selectively inhibits the synthesis of large and highly charged chondroitin/dermatan sulfate proteoglycans (CS/DSPGs). However, the size of chondroitin/dermatan sulfate chains of the CS/DSPGs was M(r) approximately 47000 in both the control and lead-treated cultures. On the other hand, lead decreased the accumulation of a large CS/DSPG with a core protein of approximately 450 kDa in the cell layer and the conditioned medium; the core protein was identified as versican core by Western blot analysis. It is therefore suggested that lead inhibits the synthesis of the versican core protein in vascular smooth muscle cells without a change in length of chondroitin/dermatan sulfate side chains. As a result, versican-poor extracellular matrix would be induced by lead in vascular smooth muscle cells.  相似文献   

13.
Pericytes associate with the outside of endothelial cells in microvessels. Previous studies have shown that these cells synthesize glycosaminoglycans (GAGs) but the nature of the core proteins to which these GAGs are attached is unknown. In the present study, cultured bovine retinal pericytes were metabolically labeled with [(3)H]glucosamine, [(35)S]sodium sulfate or (35)S-labeled amino acids and the proteoglycans synthesized by these cells were purified by DEAE-Sephacel ion exchange and molecular sieve Sepharose CL-4B chromatography. Separated proteoglycans were digested with papain, heparitinase or chondroitin ABC lyase and the GAGs characterized by Sepharose CL-6B chromatography. Proteoglycans were also assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis before and after digestion with chondroitin ABC lyase. Pericytes predominantly synthesize and secrete chondroitin or dermatan sulfate proteoglycans (CS/DS PGs) rather than heparan sulfate proteoglycans (HSPGs). Two subclasses of CS/DS PGs are synthesized by pericytes; one is a high M(r) subclass with high charge density. This subclass eluted at the void volume of a Sepharose CL-4B molecular sieve column, was susceptible to chondroitin ABC lyase, and contained core proteins of ca. 550 and 450 kD which were recognized by antibody to versican. The other major subclass eluted at a K(av) ca. 0.45 on a Sepharose CL-4B molecular sieve column, was susceptible to chondroitin ABC lyase, and contained core proteins recognized by antibodies to either biglycan or decorin that separated as a broad band of ca. 50 kDa in SDS-PAGE. A small amount of HSPG was also synthesized by these cells and could be separated from the CS/DS PGs by DEAE-Sephacel chromatography using a linear gradient of 0.1-0.7 M NaCl. Release of GAG chains by protease digestion indicated that the length of GAG chains was approximately M(r) 45000 in biglycan and decorin, approximately M(r) 48000 in the small amount of HSPGs and approximately M(r) 66000 in versican. These proteoglycans resemble those synthesized by vascular smooth muscle cells but differ markedly from those synthesized by vascular endothelial cells.  相似文献   

14.
The antitumor activity of the sea snake venom (Lapemis curtus) was evaluated against Ehrlich's ascites carcinoma (EAC) in Swiss albino mice and HeLa and Hep2 tumor cell cultures. Among the different doses tested, 6.50 microg/ml at 24 h was found to effectively inhibit cancer cell proliferation. The same dose on EAC-bearing mice by i.p. injection significantly reduced the tumor growth and was demonstrated by increased life span of the mice by 201.25%.  相似文献   

15.
Glycosaminoglycans (GAGs) are the most abundant group of heteropolysaccharides found in the body. These long unbranched molecules contain a repeating disaccharide unit. GAGs are located primarily in the extracellular matrix or on the surface of cells. These molecules serve as lubricants in the joints while at the same time providing structural rigidity to cells. Sulodexide is a highly purified glycosaminoglycan composed of a fast mobility heparin fraction as well as dermatan sulfate. Sulodexide differs from other glycosaminoglycans, like heparin, by having a longer half-life and a reduced effect on systemic clotting and bleeding. In addition, sulodexide demonstrates a lipolytic activity that is increased in comparison to heparin. Oral administration of sulodexide results in the release of tissue plasminogen activator and an increase in fibrinolytic activities. An increasing body of research has demonstrated the safety and efficacy of sulodexide in a wide range of vascular pathologies.  相似文献   

16.
Sodium spirulan (Na-SP) is a sulfated polysaccharide isolated from the blue-green alga Spirulina platensis, which consists of two types of disaccharide repeating units, O-hexuronosyl-rhamnose (aldobiuronic acid) and O-rhamnosyl-3-O-methylrhamnose (acofriose) with sulfate groups, other minor saccharides and sodium ion. Vascular endothelial cells are present on the inner surface of blood vessels in a monolayer and have anticoagulant properties. To address the question whether Na-SP influences the maintenance of endothelial cell monolayers, we investigated the proliferation of cultured bovine aortic endothelial cells treated with Na-SP. It was found that Na-SP has an inhibitory activity on endothelial cell proliferation accompanied with suppression of whole protein synthesis but without non-specific cell damage. The inhibitory activity of Na-SP was the strongest when compared to that of heparan sulfate, heparin, dextran sulfate, dermatan sulfate, chondroitin sulfate A/C and hyaluronan. Furthermore, it was shown that the inhibitory activity of Na-SP disappeared by either desulfation or depolymerization. The present data suggest that Na-SP is a unique sulfated polysaccharide that strongly inhibits vascular endothelial cell proliferation, and the inhibitory activity requires polymerization of sulfated O-rhamnosyl-acofriose repeating units.  相似文献   

17.
Capillary electrophoresis (CE) was applied to the quantitation of dermatan sulfate (DS) and chondroitin sulfate (CS) as related substances in sodium heparin. The method is based on the selective digestion of either CS and DS contained in the main drug heparin, by using chondroitinase ABC (specific for both DS and CS) and chondroitinase AC (specific for only CS). The unsaturated disaccharides released after exhaustive digestion, can be separated by CE using a 110 mM phosphate buffer, pH 3.5 as the background electrolyte in a fused silica capillary (64.5 cm × 50 μm i.d.) at 40 °C and −30 kV. Since the level of each disaccharide released upon enzymatic digestion corresponds to its content in the native glycosaminoglycan, the amount of CS and DS was determined by proportion with the released disaccharides. In particular, ΔUA → GalNAc-4S Na2 and ΔUA → GalNAc-6S Na2 were selected for quantitation of CS and DS because of their significant response and short migration time (less than 7 min).The method was validated for linearity, accuracy, precision and it showed to be able in detecting selectively, DS and CS at impurity level (LOD 0.01%, w/w). The proposed CE approach was finally applied to real samples. The results obtained were found in excellent correlation with those achieved by the analysis of the same samples using the official USP method based on high performance anion exchange chromatography (HPAEC) with pulsed amperometric detector.  相似文献   

18.
Human plasma kallikrein (huPK) is a serine proteinase involved in many biological processes including those of the kallikrein-kinin system. The action of huPK on kininogen results in bradykinin (BK) release, a potent mediator of inflammatory responses. BK generation may be influenced by several agents, and the aim of this work was to investigate the effect of glycosaminoglycans (GAGs) on human high-molecular-weight kininogen (HK) hydrolysis by huPK and on inflammation. huPK was pre-incubated in the absence and presence of different GAGs, followed by the addition of kininogen. Bradykinin released at different times was measured by radioimmunoassay, and KM and kcat were calculated. Tuna and bovine dermatan sulfates, the most potent GAGs studied, reduced by 80% and 68%, respectively, the catalytic efficiency of huPK (control = 4. x 10(4) M(-1) s(-1) in BK release. The effect of bovine dermatan sulfate (BDS) on inflammatory response was studied in rat paw edema induced by carrageenin and hourly determined (1-4 h) by plethysmography. BDS significantly reduced the inflammatory response in the first and second hours of measurements (24% and 28%, respectively), p < 0.05. GAGs were shown to reduce bradykinin release "in vitro" and in an inflammation model. This reduction may play a role in the control or maintenance of some pathological and physiological processes.  相似文献   

19.
Proteoglycans (PGs), especially chondroitin/dermatan sulfate proteoglycans (CS/DSPGs), accumulate and their composition variously changes in atherosclerotic vascular walls. Since cadmium causes atherosclerosis in experimental animals, PGs synthesized by cultured vascular smooth muscle cells after exposure to cadmium were characterized in the present study. Sparse and dense cultures of the cells were metabolically labeled with [35S]sulfate for 24 h in the presence of cadmium chloride at noncytotoxic levels (0.2 microM or less). The incorporation of [35S]sulfate into glycosaminoglycans was determined by the cetylpyridinium chloride precipitation method. The labeled PGs were characterized by DEAE-Sephacel ion exchange chromatography and Sepharose CL-4B molecular sieve chromatography. The M(r) and the glycosaminoglycan composition of small CS/DSPGs were analyzed by SDS-polyacrylamide gel electrophoresis and Sepharose CL-6B chromatography, respectively, before and after digestion with chondroitin ABC lyase or papain. The core proteins were identified by Western blot analysis. These experiments indicate that cadmium differentially acts on the PG synthesis when vascular smooth muscle cell density is low. Specifically, cadmium increased the accumulation of small CS/DSPGs identified as biglycan and decorin in the cell layer of sparse cells. However, the hydrodynamic size and the length of chondroitin/dermatan sulfate chains in the PGs were unaffected by cadmium. On the other hand, cadmium decreased other cell layer-associated PGs that were separated from biglycan and decorin by DEAE-Sephacel chromatography in the sparse cells; as the result, whole glycosaminoglycans were decreased in both the cell layer and the conditioned medium. It is therefore concluded that cadmium may change the composition of PGs in atherosclerotic plaques through induction of biglycan and decorin synthesis and inhibition of other PG synthesis in vascular smooth muscle cells.  相似文献   

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