坛紫菜硫酸多糖对60Co辐射引起的小鼠体内氧化应激的作用研究

应用生物化学的方法探讨在^60Co辐射条件下，坛紫菜硫酸多糖对小鼠体内氧化应激的抑制作用。结果表明，^60Co辐射能显著降低昆明种小鼠体内不同脏器组织匀浆SOD、GSH—Px的活性，引起脂质过氧化。坛紫菜硫酸多糖可提高辐射损伤后小鼠组织匀浆SOD、GSH—Px的活性，增强总抗氧化能力，减少脂质过氧化。     

紫菜多糖对免疫低功小鼠脾细胞功能的影响

目的探讨紫菜多糖对免疫低功小鼠脾细胞功能的影响。方法小鼠腹腔注射不同剂量(0.025,0.050和0.150g·kg-1)的紫菜多糖,每日1次,连续7d,并在给予紫菜多糖的第1天和第2天同时注射环磷酰胺造成免疫功能低下模型,第8天检测小鼠脾淋巴细胞的增殖及TNF的生物学活性。结果0.150g·kg-1紫菜多糖组小鼠的淋巴细胞增殖能力及TNF的细胞毒活性均高于模型组。结论适宜浓度的紫菜多糖能够促进淋巴细胞增殖,提高TNF的产生。     

紫菜多糖的免疫功能增强作用

目的探讨紫菜多糖对免疫功能低下小鼠的保护作用。方法小鼠腹腔注射不同剂量(0.025,0.050和0.150g.kg-1)的紫菜多糖,每日1次,连续7d,并在给予紫菜多糖的第1d,2d同时注射环磷酰胺造成免疫功能低下模型,第8d检测小鼠的NK细胞细胞毒活性,以及脾细胞分泌IFN-γ和NO的水平。结果0.150g.kg-1紫菜多糖组小鼠的NK细胞毒活性、脾细胞分泌IFN-γ及NO的水平均高于模型组。结论紫菜多糖具有一定的免疫功能增强作用。     

苗药黑骨藤多糖部位HGT-5A的细胞免疫抑制作用及其可能的活性成分

目的观察苗药黑骨藤多糖部位HGT-5A的细胞免疫抑制作用及探讨其可能的活性成分。方法采用二硝基氯苯建立小鼠迟发型超敏反应模型,初次致敏当天开始ig给予HGT-5A,每天1次,连续10 d,第11天处死小鼠测定耳肿胀度,观察HGT-5A体内对细胞免疫反应的影响;用[3 H]TdR掺入法检测HGT-5A及其多糖组分对小鼠脾细胞增殖反应的影响;用MTT法检测HGT-5A对脾淋巴细胞存活的影响。结果HGT-5A 50和100 mg.kg-1可以明显抑制迟发型超敏反应模型小鼠耳肿胀,耳肿胀度由模型组的(8.9±2.2)mg分别降低至6.4±1.7和(7.1±1.5)mg;HGT-5A 50～500 mg.L-1体外应用可促进小鼠脾细胞自发增殖反应,但在HGT-5A 100 mg.L-1以上时可明显抑制刀豆蛋白A诱导的小鼠脾细胞增殖反应(P<0.05),对脂多糖诱导的小鼠脾细胞增殖反应无明显影响;HGT-5A与脾细胞共培养24,48和72 h对脾细胞存活无明显影响。从HGT-5A分离获得的中性糖部位HP1,酸性糖部位HP2,从HP1中分离得到的多糖成分HP1-3,以及从HP2中分离得到的多糖成分HP2-3和HP2-4 0.5～50 mg.L-1可促进脾细胞自发增殖反应;HP2,HP1-3,HP1-4,HP2-2和HP2-4明显抑制刀豆蛋白A诱导的T细胞增殖反应。结论 HGT-5A可抑制T细胞活化增殖,对细胞免疫反应具有抑制作用,多糖成分HP1-3,HP1-4,HP2-2和HP2-4可能是黑骨藤发挥免疫抑制作用的活性成分。     

几种茶叶多糖对佐剂性关节炎大鼠免疫指标的影响

目的 研究几种不同途径提取的茶叶多糖(TPS)对佐剂性关节炎(AA)大鼠免疫指标的影响。方法 采用佐AA大鼠脾淋巴细胞和疆腔巨噬细胞体外培养技术研究几种TPS对其功能的影响。结果 几种茶叶多糖都有提高AA大鼠过低的脾淋巴细胞增殖反应和IL-2的趋势，其中P2途径提取的TPS对脾淋巴细胞增殖厦应和IL-2具有明显的促进作用(P<0．05)；而对AA大鼠腹腔巨噬细胞产生过高的IL-1，几种不同途径提取的TPS均有降低趋势，以P2茶叶多糖的作用最显(P<0.05)。结论 P：TPS对AA大鼠免疫指标具有明显的调节作用。     

珠蚌多糖对实验性移植肿瘤及NK细胞活性的作用

目的研究珠蚌多糖对实验性移植小鼠肿瘤以及对自然杀伤细胞（NK细胞）活性的影响。方法采用两种小鼠移植性肿瘤模型，观察珠蚌多糖对在体肿瘤细胞生长的影响；通过脾淋巴细胞与K562肿瘤细胞的共培养，体外检测NK细胞的活性。结果珠蚌多糖200，100mg·kg^-1对小鼠S180肉瘤的抑制率分别达到49．4％和47．1％，对C57BL／6小鼠B16BL6黑色素瘤的抑瘤率分别为39．1％和34．2％；显著提高S180肉瘤小鼠免疫脏器胸腺指数、脾指数；体外10，100μg·mL^-1珠蚌多糖可增强NK细胞对K562细胞的抑制活性。结论珠蚌多糖对实验移植性小鼠肿瘤生长有明显的抑制作用，体外一定剂量范围内可诱导NK细胞活性。     

中华眼镜蛇毒组分F的体外抗血管生成活性研究

目的研究中华眼镜蛇毒组分F对血管内皮细胞黏附功能的抑制作用和体外的抗血管生成活性。方法MTT快速测定法测定组分F对原代培养的血管内皮细胞对纤维蛋白原(Fg)、纤连蛋白(Fn)、Matrigel的黏附作用的影响;采用平面拟毛细血管生成培养法,观察组分F对血管生成的抑制效应。结果组分F可抑制血管内皮细胞的黏附功能,对细胞黏附Fn、Fg和Matrigel 3种物质的作用从1.2μg/ml浓度起即有抑制效应(P<0.05),且均呈浓度依赖性,组分F对细胞黏附Fg和Matrigel的抑制作用较其对Fn的作用强(P<0.05);组分F可抑制内皮细胞在培养基质中生成血管样网状结构的反应,并且随浓度的不同抑制程度有明显不同。结论中华眼镜蛇毒组分F具有体外抗血管生成活性。     

枸杞多糖对肝癌Hca-F 荷瘤小鼠的抗肿瘤作用及其机制

目的研究枸杞多糖（LBP）对小鼠Hca-F肝癌的生长抑制作用及机制。方法建立小鼠Hca-F肝癌模型,腹腔注射不同浓度的枸杞多糖连续10d。称重检测枸杞多糖对肿瘤生长、脾脏和胸腺指数的影响;采用MTT法、ELISA法分析枸杞多糖对荷瘤小鼠脾巨噬细胞的吞噬活性、细胞因子分泌等免疫指标的影响。结果枸杞多糖对小鼠实体瘤生长具有抑制作用,枸杞多糖40mg·kg^-1与对照组相比有显著差异（P〈0.05）;实验组小鼠脾指数和胸腺指数均高于对照组。枸杞多糖40mg·kg^-1即可明显促进IL-2的产生,降低VEGF蛋白的表达。结论枸杞多糖对Hca-F肝癌小鼠肿瘤生长有抑制作用,可能与提高荷瘤小鼠细胞和分子免疫活性有关。     

黄丹兰颗粒剂对肝癌Hca-F荷瘤小鼠的抗肿瘤作用及其机制

目的：研究黄丹兰颗粒剂对小鼠 Hca - F 肝癌的生长抑制作用及机制。方法：建立小鼠Hca - F肝癌模型,口服灌胃不同浓度的黄丹兰颗粒剂连续10 d。称重检测黄丹兰颗粒剂对肿瘤生长、脾脏和胸腺指数的影响;采用 ELISA 法分析黄丹兰颗粒剂对荷瘤小鼠脾巨噬细胞的吞噬活性、细胞因子分泌等免疫指标的影响。结果：黄丹兰颗粒剂对小鼠实体瘤生长具有抑制作用,黄丹兰颗粒剂中剂量组与对照组相比有显著差异（P ﹤0.05）;实验组小鼠脾指数和胸腺指数均高于对照组。黄丹兰颗粒剂中剂量组即可明显促进 IL -2的产生,降低 VEGF 蛋白的表达。结论：黄丹兰颗粒剂对 Hca - F肝癌小鼠肿瘤生长有抑制作用,可能与提高荷瘤小鼠细胞和分子免疫活性有关。     

基于多重细胞因子释放的中药活性多糖筛选和抗肿瘤作用研究

目的 肿瘤微环境的细胞因子对肿瘤进展具有重要作用。本研究拟以细胞因子释放为切入点筛选具有抗肿瘤作用的中药活性多糖。方法 选取20种中药多糖,采用Cytometric Bead Array (CBA)结合流式细胞技术筛选活性多糖;采用小鼠B16F10黑色素瘤荷瘤模型观察活性多糖的体内抗肿瘤作用。结果 17种中药多糖能显著刺激脾细胞分泌TNF-α,5种多糖能刺激脾细胞分泌IL-6;红花多糖和山慈姑多糖还能显著刺激脾细胞分泌IFN-γ。红花多糖能抑制黑色素瘤B16F10细胞在小鼠体内生长。结论 基于多重细胞因子的测定可以作为免疫抗肿瘤中药的筛选手段。红花多糖具有较好的免疫刺激作用和抗肿瘤作用。     

Extension of life span and improvement of vitality of Drosophila melanogaster by long-term supplementation with different molecular weight polysaccharides from Porphyra haitanensis.

Polysaccharides isolated from Porphyra (porphyran) have been known to have diverse biological activities, including immunomodulatory and antioxidant activities. The molecular weight-antiaging activity relationship of degraded porphyrans was examined in this study. Natural porphyran was extracted from P. haitanensis, and then was degraded into different molecular weight fractions, P1 molecular weight 49 kDa, P2 molecular weight 30 kDa, P3 molecular weight 8.2 kDa, by free radical. The influence on life span and vitality of porphyrans were carried out on Drosophila melanogaster. We found that all the degraded porphyrans and natural porphyran (P), added daily to the diet, can significantly increase the life span of D. melanogaster, except for P3. Among them, P1 exhibited the most prolonging life span activity. Furthermore, vitality of middle-aged flies (assessed by measuring their mating capacity) receiving porphyrans was increased considerably in comparison with the controls. Finally, in the heat-stress test, we observed a remarkable increase in survival time, especially in P3-diet groups. These results suggest that porphyrans may be effective in reducing the rate of the aging process and molecular weight has important influence on the effects. It seems that P1 and P2, possessed higher molecular weight, may be more useful in normal metabolic condition and P3, possessed the lowest molecular weight, may be more beneficial for D. melanogaster in stress condition.     

In vivo antioxidant activity of polysaccharide fraction from Porphyra haitanesis (Rhodephyta) in aging mice.

Sulfated polysaccharide fraction F2 from Porphyra haitanesis (Rhodephyta) showed inhibitory effect on the in vitro lipid peroxidation. In the present study, the age-related changes in the antioxidant enzyme activity, lipid peroxidation, and total antioxidant capacity (TAOC) in different organs in mice were investigated and the in vivo antioxidant effect of F2 in aging mice was checked. Increased endogenous lipid peroxidation and decreased TAOC were observed in aging mice. Intraperitoneal administration of F2 significantly decreased the lipid peroxidation in a dose-dependent manner. F2 treatment increased TAOC and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in all the organs tested in aging mice. It is concluded that the sulfated polysaccharide fraction F2 can be used in compensating the decline in TAOC and the activities of antioxidant enzymes and thereby reduces the risks of lipid peroxidation.     

Anti-adhesive effect of an acidic polysaccharide from Aloe vera L. var. chinensis (Haw.) Berger on the binding of Helicobacter pylori to the MKN-45 cell line

Objectives The emergence of antibiotic‐resistant Helicobacter pylori strains has necessitated a search for alternative therapies for the treatment of this infection. The aim of this study was to evaluate whether or not polysaccharide fractions from Aloe vera are effective in inhibiting the adherence of H. pylori in vitro. Methods Polysaccharide fractions were extracted from A. vera and subjected to carbohydrate analysis. The adhesive effect was determined by co‐incubation of H. pylori and cells with polysaccharides followed by fluorescein isothiocyanate labelling and Gram staining in vitro. Inhibition of H. pylori growth and cellular viability was tested by agar diffusion and MTT assay. Key findings APS‐F2 contained significant amounts of galacturonic acid, galactose and arabinose. APS‐F1 was galacturonic acid‐free and consisted of mannose, glucose and galactose. APS‐F2 (0.1, 0.5 and 1.0 mg/ml) reduced the count of H. pylori attached to MKN45 cells to 88, 76 and 64%, respectively. APS‐F1 did not show the same effect. Neither polysaccharide revealed an inhibitory effect on the growth of H. pylori or cell viability. In addition, APS‐F2 was shown to have a potent anti‐adhesive effect against Escherichia coli. Conclusions The results show that the acidic polysaccharide from A. vera has a potent anti‐adhesive effect against H. pylori in vitro. However, there have yet to be any in‐vivo studies to demonstrate the clinical relevance of this finding.     

红藻藻蓝蛋白对HL-60细胞生长的抑制作用

本文运用半固体琼脂培养法及 MTT法研究了红藻藻蓝蛋白 (R- PC)对 HL- 60细胞生长的影响 ,结果发现 :R- PC能够显著抑制细胞的生长 ,且存在浓度效应和时间效应关系。为进一步探索 R- PC抑制 HL- 60细胞生长的可能机制 ,将不同浓度的 R- PC与 HL- 60细胞相互作用 6d后 ,分别用流式细胞仪检测 HL- 60细胞所处细胞周期中的时相和免疫印渍法测定 HL- 60细胞中 Bcl- 2基因表达产物的变化 ,实验结果显示 :R- PC确能使更多的 HL- 60细胞滞留于细胞周期的 G0 - G1 期 ,但与 R- PC浓度无正向关系 ,且 HL- 60细胞中 Bcl- 2基因表达产物的量也没有明显的变化。结果表明 :R- PC可能通过不同于 G0 - G1 期滞留及 Bcl- 2基因表达的机制抑制 HL- 60细胞的生长。     

Free radical scavenging and apoptotic effects of Cordyceps sinensis fractionated by supercritical carbon dioxide.

Supercritical carbon dioxide (SC-CO2) was used as the elution solvent for fractioning ethanolic extract (E) of Cordyceps sinensis (CS), a traditional Chinese herbal remedy, into R, F1, F2, and F3 fractions. This extractive fractionation method is amenable to large scale and is nontoxic. These four fractions were characterized in terms of total polysaccharides and cordycepin concentrations, scavenging ability of free radicals, and anti-tumor activities. Experimental results demonstrated that fractionation altered the distributions of total polysaccharides and cordycepin in fractions. Fraction R was the most active fraction to scavenge free radicals and inhibit the proliferation of carcinoma cells, followed by the fraction F1 and the extract E. The effect of scavenging on 1,1-diphenyl-2-picryl hydrazyl (DPPH) of CS extract and fractions at 2 mg/ml was R (93%), F1 (75%), E (66%), F2 (47%), and F3 (27%). The IC50 (50% cell growth inhibitory concentration) of tumor cell proliferation and colony formation on human colorectal (HT-29 and HCT 116) and hepatocellular (Hep 3B and Hep G2) carcinoma cells by fraction R were around 2 microg/ml. Conversely, R did not affect the growth of normal dividing human peripheral blood mononuclear cells (PBMC) by exhibiting a large value of IC50 over 200 microg/ml. Accumulation of tumor cells at sub-G1 phase and the fragmentation of DNA, typical features of programmed cell death, were observed in a time and dose dependent manner. Scavenging of free radicals and anti-cancer activity (value of IC50) correlated closely with the quantities of polysaccharides (Spearman's rho=0.901 and -0.870, respectively). Taken together, our findings suggest that fraction R, obtained by SC-CO2 fluid extractive fractionation, showed strong scavenging ability and selectively inhibited the growth of colorectal and hepatocellular cancer cells by the process of apoptosis.     

半边旗提取物6F抑制HL-6 0细胞蛋白激酶C活性(英文)

目的　探讨半边旗提取物 6F的细胞毒作用及诱导DNA片段化与蛋白激酶C(PKC)信号转导途径的关系 ,检测 6F对PKC活性的影响。方法　受试对象为HL 6 0细胞 ,超速离心法获得的胞液 (可溶部分 )及颗粒 (不可溶部分 ,包括细胞膜系统及胞核 )部分用作PKC活性测定。经 0 .4g·L- 1磷脂酰丝氨酸 ,0 .0 4g·L- 1甘油二油酸酯激动剂作用酶粗提物后 ,用液体闪烁计数仪计数 [γ 32 P]ATP参入外源底物的量以测定PKC活性。MTT法测定HL 6 0细胞的活力 ,二苯胺法测定 6F诱导DNA片段化程度。结果　在所测试的浓度范围内 (0 .5～ 312 μmol·L- 1) ,化合物 6F显著抑制胞液及颗粒部分PKC活性 ,最大抑制率达 88.6 % ,呈浓度依赖关系 (胞液部分r =0 .781,P <0 .0 5 ,颗粒部分r =0 .931,P <0 .0 1)。 6F诱导HL 6 0细胞DNA片段化及对细胞的毒性作用可被具有致癌作用的PKC激活剂肉豆蔻酸酯 (PMA ,浓度为 6 5nmol·L- 1)拮抗 ,抑制率分别是 30 %和 4 4% (P <0 .0 1)。PMA单独用使HL 6 0细胞线粒体将MTT还原为甲月赞的能力增强 14 % (P <0 .0 1) ,即增强细胞活力。结论化合物 6F是PKC的抑制剂。 6F对HL 6 0细胞DNA片段化的诱导作用及其细胞毒作用至少可部分归因于其对PKC活性的抑制作用     

Modulation of T cells by cyclic AMP in murine influenza virus infection. I. In vitro inhibition of cytotoxic T lymphocytes by exogenous cyclic AMP analogues and by agents which increase intracellular cyclic AMP concentrations

The effects of 8-bromo-cyclic AMP and cyclic AMP agonists (cholera toxin plus hydrocortisone and prostaglandin E2 plus 3-isobutyl-1-methylxanthine) on the cytotoxic activity of T cells generated during murine influenza virus infection have been examined. Treatment of influenza A/WSN virus primed primary immune spleen cells and their cultured secondary effector T cells from Balb/c mice with these agonists resulted in increases in cyclic AMP levels. These agents also had a marked inhibitory effect on the cytotoxic activity of primary immune spleen cells, but a much weaker inhibitory effect on that of secondary effector T cells. No significant effect of 8-bromo-cyclic GMP on the cytotoxic activity of these two cell types was observed. The cytotoxic activity of H-2-restricted, virus-specific T cells generated either in vivo (primary) or in vitro (secondary) was not significantly inhibited by treatment with histamine, whereas that of alloreactive killer T cells generated in vivo was markedly inhibited. These results suggest that the actions of H-2-restricted, virus-specific T cells may not be regulated by histamine during viral or inflammatory processes.     

Antimicrobial activity of flavonoids from Pelargonium radula (Cav.) L'Hérit

Flavonoids from Pelargonium radula (Cav.) L'Hérit were purified by column chromatography. Two fractions were obtained: F1 (main flavonoid isoquercitrin) and F2 (main flavonoid rutin). In vitro antimicrobial activity of F1 and F2 were tested against eleven species of bacteria and eleven species of fungi. Both fractions demonstrated strong inhibitory activity against Staphylococcus aureus, Proteus rettgeri, Candida tropicalis and Microsporum gypseum. Staphylococcus sp. (coagulase-negative) and Candida lusitaniae were strongly inhibited only by fraction F1 and Fusarium graminearum only by fraction F2.     

Inhibition of Helicobacter pylori hemagglutination by polysaccharide fractions from roots of Panax ginseng

Polysaccharides from the roots of Panax ginseng were extracted by hot water and fractionated by using ethanol precipitation and ion exchange chromatography. Fractions FC (crude extract), F1 (fraction precipitated by ethanol), F1N (fraction unbound to DEAE-Sepharose CL-6B), and F1A (bound fraction) were obtained. Their carbohydrate analyses showed that acidic fraction F1A contains higher amounts of galactose, arabinose and uronic acids, in comparison to FC and F1. Fraction F1N mainly consists of glucose. The inhibition of Helicobacter pylori-induced hemagglutination revealed different inhibitory activities of these fractions. In particular, acidic fraction F1A showed a remarkable inhibitory activity (minimum inhibition concentration was 0.25 mg/ml) among the polysacharide fractions. However, digestion of the fraction F1A with pectinase resulted in a lower molecular weight oligosaccharide fraction F1AP which was non-inhibitory at the concentration of 4 mg/ml. Comparison of inhibitory activities and carbohydrate compositions of isolated fractions indicates that the activity correlated with the contents of galactose, arabinose, and uronic acids. These data suggest that acidic polysaccharides may be responsible for the inhibitory activity.