Structural characterization of three acidic polysaccharides from Opuntia dillenii Haw.fruits and their protective effect against hydrogen peroxide-induced oxidative stress in Huh-7 cells

Three novel acidic polysaccharide fractions(OFPP-1,OFPP-2,OFPP-3)with different m olecular weights(803.7,555.1 and 414.5 k Da)were isolated from the peeled Opuntia dillenii Haw.fruits by alkali-extraction,graded alcohol precipitation and column chromatography.Structural analysis indicated that OFPPs were pectic polysaccharides consisting of rhamnose,arabinose and galactose residues.The backbone of OFPP-1 consisted of a repeating unit→6-α-D-Galp A-(1→2)-α-L-Rhap-(1→with T-α-D-Galp A-(1→6)-α-D-Galp A-(1→4)-α-D-Glcp-(1→,T-β-D-Xylp-(1→6)-α-D-Galp A-(1→4)-α-D-Glcp-(1→or T-α-D-Galp A-(1→3)-α-L-Araf-(1→as the side chains.The backbone of OFPP-2 consisted of a disaccharide repeating unit→2)-α-L-Rhap-(1→4)-β-D-Galp A-(1→with T-β-L-Araf-(1→as the branches substituted at the O-4 position of→2,4)-α-LRhap-(1→.Whereas the backbone of OFPP-3 was→2,4)-α-L-Rhap-(1→2)-α-L-Rhap-(1→3)-β-L-Araf-(1→or→2,4)-α-L-Rhap-(1→2)-α-L-Rhap-(1→4)-β-D-Galp A-(1→,which was branched at the O-4 position of→2,4)-α-L-Rhap-(1→.Moreover,these three polysaccharide fractions could protect Huh-7 cells against H2O2-induced oxidative stress to different extents by decreasing the MDA content and increasing the SOD,CAT,GSH-Px activities and the GSH level in the Huh-7 cells.These results suggest that OFPPs have the potential to be used as natural antioxidants.

Anti-oxidation Activity in Vitro of Polysaccharides of Dendrobium Huoshanense and Dendrobium moniliforme

[ Objective ] The aim was to study the effects of polysaccharides of Dendrobium Huoshanense and Dendrobium moniliforme on oxygen free radicals and lipid peroxidation in vitro. [ Method] The scavenging action on .OH of polysaccharides, the inhibition action on O2^- of polysaccharides, MDA production of lipid peroxidation were measured by Fenton reaction, self oxidation system of C6H3（ OH）3 and TBA method respectively. [ Result] The half scavenging concentration on ·OH of potysaccharides of Dendrobium Huoshanense and Dendrobium moniliforme was 6.79 and 6.75 mg/ml respectively. As to their half inhibition concentration on O2 , the data was 3, 04 and 3.44 mg/ml respectively. Two kinds of Dendrobium polysaccharides bothhad inhibitory actions on lipid peroxidation of liver homogenate induced by auto-oxidation and inducedoxidation, and they could alleviate oxidation damages of mice liver mitochondria induced by Vc-Fe2＋ system. [Conclusion] Two kinds of Dendrobium polysacchaddes both had remarkable anti-oxidation activity in vitro.

Roles of polysaccharide from Branchiostoma belcheri in anti-DNA oxidation and anti-tumor activity in S180 mice

In this study, we isolated a polysaccharide from Branchiostoma belcheri （PBB） by enzymatic protein hydrolysis and alcohol precipitation. We investigated the effects of PBB supplementation on DNA oxidation and growth of the transplanted tumor cells Sarcoma （S180） in mice. Sixty healthy Kunming mice weighing between 18 and 25 g were randomly assigned to 6 groups, each consisting of 10 animals. All the mice, except for the blank control group, were inoculated with S180 sarcoma cells into the axilla of the left foreleg. PBB was given to mice by gavage at doses of 0 （model control）, 25, 50, or 100 mg/kg b.w. in 0.2 ml saline for 30 days. The fifth group of S180-mice was given cytoxan （50 mg/kg） by peritoneal injection as a positive control group. The animals had free access to food and water. The mice were sacrificed after the final treatment and blood was quickly collected. Spontaneous and oxidized DNA damage of peripheral lymphocytes induced by H2O2 were analyzed by SCGE. O6-methyl-guanine （O6-MeG） was measured by high-performance capillary zone electrophoresis. The average tumor weights （0.856-1.118 g） of the three PBB groups were significantly lower than that of the model control group （1.836 g） （p〈0.05）. The tumor inhibition ratios of the PBB groups were 39.1%-53.4% and similar to the cytoxan positive group （57.5%）. There were no significant differences in spontaneous DNA damage in peripheral lymphocytes among the groups. The oxidative DNA damage induced by 10 μmol/L H2O2 in the 50 and 100 mg/kg b.w. groups were 246.1 AU and 221.7 AU, respectively, both of which were significantly lower than that in the model group （289.0 AU; p〈0.05）. The plasma concentrations of O6-MeG in the 25, 50, and 100 mg/kg supplemented groups were 2.09 μmol/L, 1.86 μmol/L, and 1.63 lamol/L, respectively, all of which were significantly lower than that of the model group （2.67 μmol/L; p〈0.05）. These results indicated that PBB may have antioxidative activity and thus reduce oxidation-induced DNA damage.

Polysaccharide extracts of Cirsium japonicum protect rat H9c2 myocardial cells from oxidative stress induced by hydrogen peroxide

Daji （Cirsium japonicum） has been applied against gastric disorders, lung diseases, and cardiovascular problems in thetraditional Chinese medicinal system. The present study was to investigate the protective effects of Daji （Cirsiumjaponicum） polysaccharide extracts （CJP） against hydrogen peroxide （H2O2） shock in rat H9c2 myocardial cells. First,CJP was isolated by hot water extraction and ethanol precipitation; it was then characterized by high performance liquidchromatography and infrared spectrum analysis. Rat H9c2 cells were subjected to H2O2 treatment to establish a cellinjury model. The 3- （4,5- dimethylthiazol- 2-yl）-2,5- diphenyltetrazolium bromide assay showed that CJP pretreatmentsignificantly ameliorated the H2O2 injury in a dose-dependent manner. Furthermore, the cell apoptosis induced by H2O2was markedly inhibited by CJP pretreatment, whereas the cleavage level of caspase-3, -8, and -9 was reduced. Inaddition, the p38 mitogen-activated protein kinase pathway might be involved in the protective effect of CJP onmyocardial cells. Therefore, we conclude that polysaccharide extracts of Daji （Cirsium japonicum） protect rat H9c2myocardial cells from oxidative stress induced by H2O2.

响应面法优化云南华坪仙人掌多糖提取工艺及抗癌活性研究

(目的)探讨仙人掌中多糖的提取工艺和抗癌活性。(方法)利用单因素实验和响应面法优化了仙人掌粗多糖的提取工艺,并利用MTT法评价该多糖的抗癌活性。(结果)经响应面法优化得到的仙人掌粗多糖的最佳提取工艺为:提取时间5 h、提取次数5次、料液比1∶60 g/mL、乙醇体积分数70%,仙人掌粗多糖提取率为87.02%。经响应面预测得到的最佳工艺与实验条件最佳条件相同,经进一步验证提取率为(82.38±3.19)%。MTT法测定仙人掌粗多糖抑制肿瘤细胞生长活性的结果表明,仙人掌粗多糖对结肠癌细胞(RKO)和乳腺癌细胞(4T1)有一定的抑制活性,IC_(50)值分别为143.5和167.9 mg/L。(结论)本研究获得的工艺条件能有效的提取仙人掌胶中的多糖,且得率高;活性实验表明仙人掌胶多糖对结肠癌细胞(RKO)和乳腺癌细胞(4T1)有一定的抑制活性。

仙人掌多糖的分离、纯化及性质研究

目的分离、纯化具有降血糖作用的仙人掌多糖组分。方法经热水提取、乙醇沉淀、DEAE Sepharose fast flow离子交换色谱和Sephadex G系列凝胶滤过色谱纯化得到5种仙人掌多糖组分。醋酸纤维薄膜电泳检测多糖纯度,凝胶色谱测定相对分子质量,高效液相色谱测定糖的组成。结果5种多糖都已达到电泳纯。它们的相对分子质量依次为2.0×103、1.0×106、4.0×103、9.2×105、5.0×103。ODP1由鼠李糖组成,ODP2可能由鼠李糖和葡萄糖组成,ODP4可能由鼠李糖和D-半乳糖组成。ODP3和ODP5分别由鼠李糖和一未知糖组分组成。结论仙人掌多糖的提取没有必要采用脱蛋白、脱色素步骤,本实验所提取的仙人掌多糖是均一的。

仙人掌多糖的降血糖作用

为观察仙人掌多糖的降血糖作用,依次往小鼠腹腔注射链脲佐菌素120,40mg/kg,建立糖尿病模型;然后,将小鼠随机分为3组,分别灌胃仙人掌多糖150,200,250mg/kg,并测定其各项生理指标.结果表明,仙人掌多糖能明显改善糖尿病小鼠多饮、多食、消瘦症状,具有降血糖作用.

微波辅助提取仙人掌多糖的工艺研究

该文采用微波辅助法提取米邦塔仙人掌多糖,并与热回流提取方法进行比较。通过设计正交试验,得出优化的工艺条件:以水为提取剂,料液比1∶10(w/v),提取2次,每次提取3 m in,微波炉功率700 W,选用70%乙醇沉淀。微波提取粗多糖的得率和含量分别为6.8%和8.5%,高于热回流提取粗多糖的得率4.7%和含量8.3%。微波提取不仅缩短了提取时间,降低提取剂用量,而且提高了仙人掌多糖得率。

酸法提取仙人掌多糖工艺

为探索适合产业化的仙人掌多糖(Opuntia dillenii Haw.polysaccharides,ODPs)提取工艺,采用单因素及正交试验优化酸法提取ODPs条件;优化的苯酚硫酸法测定ODPs得率及含量;并用酶法结合盐酸法除蛋白,经透析,洗涤得到精制的ODPs。得到酸法提取ODPs最佳条件为:加入浓度为0.10 mol/L的硫酸溶液,液料比为20∶1(mL/g),70℃提取2次,每次2 h。平均产率达39.92%,多糖纯度达87.11%,多糖中蛋白含量降低至1.06%。

仙人掌多糖发酵提取工艺优化及其抗炎功效研究

本文分别采用乳酸菌与酵母菌发酵提取仙人掌多糖,并探究仙人掌多糖的抗炎功效。在三个单因素液料比、时间、pH方面探究仙人掌多糖的提取工艺,得到三因素最适组合,乳酸菌发酵最适提取工艺为发酵时间7 h、液料比25∶1 (mL/g)、pH为5,此工艺下多糖得率为0.26%。黄酒酵母发酵最适提取工艺为发酵时间48 h、液料比25∶1 (mL/g)、pH为3,此工艺下多糖得率为0.53%。应用实时荧光定量PCR法检测仙人掌多糖对IL-6、IL-8炎症细胞因子相关基因表达的影响,得知仙人掌多糖通过抑制IL-8的表达,促进IL-6的表达来抑制炎症的发生和发展。

野生仙人掌多糖对DPPH·、NO_2^-的清除能力及其还原力研究

利用热水浸提法提取野生仙人掌多糖,并用Sevage法对其进行纯化,通过测定DPPH.、NO2-的清除作用和其还原力,研究野生仙人掌多糖的抗氧化活性。结果表明,野生仙人掌多糖对DPPH.、NO2-有较强的清除作用,并具有一定的还原能力,其抗氧化性有待深入研究和开发应用。

野生仙人掌多糖对小鼠特异性免疫功能的调节作用

目的:观察野生仙人掌多糖(ODPs)对小鼠特异性(体液及细胞)免疫功能的调节作用。方法:采用环磷酰胺建立免疫抑制小鼠模型,检测ODPs对血清溶血素IgMI、gG含量、脾淋巴细胞增殖反应和外周血T淋巴细胞亚群比例的影响;检测分离纯化的ODPs主要组分ODP-I对体外培养的淋巴细胞增殖作用。结果:ODPs能显著增强免疫抑制小鼠血清溶血素IgMI、gG的生成;显著增强免疫抑制小鼠T、B淋巴细胞的增殖能力,且均使之恢复至正常水平;显著降低免疫抑制小鼠CD4+T细胞亚群比例,从而显著降低CD4+/CD8+T细胞比值,使之接近正常水平,且均有剂量关系。ODP-I可极显著促进小鼠脾淋巴细胞自然增殖,协同有丝分裂原ConA或LPS诱导T、B淋巴细胞的增殖。结论:ODPs对免疫抑制小鼠特异性免疫功能具有调节作用,对体外淋巴细胞具有增殖作用,是一种良好的免疫调节剂。

仙人掌多糖对小鼠急性肝损伤的保护作用

目的研究仙人掌多糖(OPS)对CCl4肝损伤小鼠的保护作用。方法采用OPS溶液预先灌胃30 d,末次灌胃后2 h,腹腔注射0.10%CCl4(20 mL/kg)的花生油溶液,造成小鼠急性肝损伤,测定血清ALT活性和AST活性及肝脏GSH含量、GR活性、GST活性和GSH-Px活性,取肝组织做病理学检测。结果预先灌胃OPS溶液,可显著地抑制CCl4引起的小鼠血清ALT和AST活性升高,改善肝中毒小鼠GSH含量及GST活性和GSH-Px活性水平,并减轻肝组织的病理变化。结论OPS对CCl4致小鼠肝损伤具有保护作用,这可能与增强谷胱甘肽系统抗氧化能力有关。

仙人掌肉质茎成分的分离与结构鉴定

从仙人掌 [Opuntiadillenii(Ker Gawl.)Haw .]肉质茎分离得到 6个化合物 ,经过理化性质和光谱分别鉴定为 :3 O 甲基槲皮素 (1)、山奈酚 (2 )、山奈甲黄素 (3)、槲皮素 (4)、异鼠李黄素(5 )、β 谷甾醇 (6 ) ,其中化合物 (1)、(2 )、(3)

仙人掌水溶性粗多糖提取及抗氧化性能研究

研究仙人掌水溶性粗多糖的提取工艺及抗氧化性能。采用单因素试验和L(933)正交试验设计,考察提取温度、时间、料液比等因素对提取率的影响,并采用水杨酸法考察仙人掌粗多糖抗氧化性能。结果表明:温度是影响粗多糖提取的重要因素。优化工艺条件为:提取温度80℃,提取时间1h,固液比为1:60。抗氧化试验表明仙人掌粗多糖有较好的抗氧化活性。

仙人掌的化学成分研究

目的:研究仙人掌的化学成分。方法:采用多种层析手段进行分离纯化,应用多种波谱技术,结合文献对照,对分离得到的化合物进行结构鉴定。结果:从仙人掌肉质茎的乙醇提取物中分离出6个化合物,其结构分别鉴定为3,4-二羟基苯甲酸乙酯(1),3,4-二羟基苯甲酸(2),槲皮素3O甲基-7-O-β-D-葡萄糖苷(3),山奈酚7OβD葡萄糖苷(4),manghaslin(5)和山奈酚-7-O-β-D-葡萄糖基(1→4)-β-D-葡萄糖苷(6)。结论:化合物6为新化合物,化合物1～5均为首次从本属植物中分离得到。

野生仙人掌粗多糖脱蛋白方法的研究

采用三氯乙酸(TCA)法、Sevag法、蛋白质等电点法对仙人掌粗多糖进行脱蛋白研究,以蛋白脱除率、多糖损失率和抗氧化性损失率为指标。结果表明:三氯乙酸法脱蛋白效果最好,浓度为3%时,蛋白脱除率为80.28%,多糖损失率为5.98%,抗氧化性损失率为34.56%。

仙人掌多糖主要组分对大鼠红细胞脂质过氧化损伤的影响

目的分离纯化仙人掌多糖主要组分,观察仙人掌多糖主要组分(ODP-Ⅰa)对红细胞脂质过氧化损伤的影响,探讨其抗氧化机制。方法从野生仙人掌Opuntia dilleniiHaw.中采用超声提取法并用DEAE-Sepharose Fast Flow柱层析纯化得到仙人掌多糖组分ODP-Ⅰa、ODP-Ⅰb和ODP-Ⅱ′。以活性氧(H2O2)作用于离体正常大鼠红细胞,观察ODP-Ⅰa对红细胞溶血率、高铁血红蛋白含量、丙二醛含量、超氧化物歧化酶和谷胱甘肽过氧化物酶活性、红细胞膜蛋白巯基含量和膜蛋白质高分子聚合物的影响。结果 ODP-Ⅰa能够抑制H2O2诱导的红细胞溶血和高铁血红蛋白的产生;提高H2O2氧化损伤红细胞中超氧化物歧化酶活性、谷胱甘肽过氧化物酶活性和降低丙二醛含量;同时,能够抑制H2O2引起的膜蛋白巯基含量的降低和红细胞膜蛋白高分子聚合物的形成。结论 ODP-Ⅰa对活性氧引起的红细胞脂质过氧化损伤具有保护作用。

仙人掌中一个新α-吡喃酮成分的分离与结构鉴定

目的 研究仙人掌的化学成分。方法 应用多种柱色谱方法进行分离和纯化 ,NMR和MS等波谱解析化学结构。结果 从仙人掌肉质茎的乙醇提取物中分离出 6个化合物 ,其结构分别鉴定为 :3 O 甲基异鼠李黄素 (1)、4 乙氧基 6 羟甲基 α 吡喃酮 (2 )、正十七醇 (3)、香草酸 (4)、异鼠李黄素 3 O 鼠李糖苷 (5 )和芦丁 (6 )。结论 化合物2为新化合物 ,化合物 1、3、4、5均为首次从本属植物中分离得到 ,化合物 6为本植物首次分离得到。

褐藻硫酸多糖清除活性氧自由基作用及动力学的ESR研究

利用ＥＳＲ和自旋捕集技术，首次观察了不同方法提取的两种褐藻硫酸多糖（ＳＰ）对Ｆｅｎｔｏｎ反应和光照Ｈ２Ｏ２两个体系产生的·ＯＨ清除作用，及对次黄嘌呤／黄嘌呤氧化酶和光照核黄素两个体系产生的Ｏ－．２（ＨＯ·２）清除作用，并且对其清除·ＯＨ和Ｏ－．２的动力学进行了研究。结果表明，热提取物（ＳＰｈ）和冷提取物（ＳＰｃ）均有明显清除ＯＨ和Ｏ－．２作用，且呈量效关系。对Ｏ－．２的清除率高于对·ＯＨ的清除率（８４．７％ｖｓ６９．２％）。ＳＰｈ对两种自由基的清除效果比ＳＰｃ好。在光照核黄素体系中ＳＰｈ３０ｇ／Ｌ几乎完全清除Ｏ－．２，ＳＰｃ可清除６８．９％，而在次黄嘌呤／黄嘌呤氧化酶体系中只需３ｇ／ＬＳＰｈ便可完全清除Ｏ－．２。１５ｇ／ＬＳＰｈ和３０ｇ／ＬＳＰｃ对Ｆｅｎｔｏｎ反应产生的·ＯＨ的清除率分别为７７．７％和６８．７％。动力学研究表明，ＳＰｈ与·ＯＨ和Ｏ－．２反应的表观速率常数分别为７．１×１０９ｍｏｌ－１·ｓ－１和３２３０．５ｍｏｌ－１·ｓ－１。实验提示ＳＰ可作为一种预防性的抗氧化剂。确证ＳＰ清除自由基为研究其抗ＨＩＶ及肿瘤的机理提供了一条新途径。