安吉白茶多糖抗肿瘤及免疫调节研究

选用MTT法研究了安吉白茶多糖的体外抗肿瘤活性;采用建立小鼠移植瘤模型,通过测定安吉白茶多糖对荷瘤小鼠抑瘤率和腹腔巨噬细胞吞噬能力,研究其体内抗肿瘤活性和免疫调节作用。结果表明,安吉白茶多糖体外能显著抑制S180细胞的生长,且具有剂量依赖关系。体内试验表明,与模型对照组相比,安吉白茶多糖对小鼠S180实体瘤和H22腹水型肝癌移植瘤的生长均有显著抑制作用(P<0.05),对荷瘤小鼠脾指数和胸腺指数也有较强增高作用。体内免疫调节试验结果显示,安吉白茶多糖可使荷S180实体瘤小鼠腹腔巨噬细胞吞噬能力显著升高(P<0.05)。     

Antioxidant and Immunity Activity of Water Extract and Crude Polysaccharide from <Emphasis Type="Italic">Ficus carica</Emphasis> L. Fruit

The antioxidative activities of water extract (WE) and crude hot-water soluble polysaccharide (PS) from Ficus carica L. fruit were investigated using various assays in vitro, including scavenging abilities on DPPH, superoxide and hydroxyl radicals and reducing power. The immunity activities of PS were evaluated using the carbon clearance test and serum hemolysin analysis in mice. In addition, total phenolics and flavonoids contents were also determined. Both WE and PS have notable scavenging activities on DPPH with the EC₅₀ values of 0.72 and 0.61 mg/ml, respectively. The PS showed higher scavenging activity than WE on superoxide radical (EC₅₀, 0.95 mg/ml) and hydroxyl anion radical (scavenging rate 43.4% at concentration of 4 mg/ml). The PS (500 mg/kg) also has a significant increase in the clearance rate of carbon particles and serum hemolysin level of normal mice. The results indicate that both WE and PS might be applicable in healthy medicine and food industry.     

Evaluation of Macroalgae Sulfated Polysaccharides on the Leishmania (L.) amazonensis Promastigote

The sulfated polysaccharides from Solieria filiformis (Sf), Botryocladia occidentalis (Bo), Caulerpa racemosa (Cr) and Gracilaria caudata (Gc) were extracted and extensively purified. These compounds were then subjected to in vitro assays to evaluate the inhibition of these polysaccharides on the growth of Leishmania (L.) amazonensis promastigotes. Under the same assay conditions, only three of the four sulfated polysaccharides were active against L. amazonensis, and the polysaccharide purified from Cr was the most potent (EC₅₀ value: 34.5 μg/mL). The polysaccharides derived from Bo and Sf demonstrated moderate anti-leishmanial activity (EC₅₀ values of 63.7 μg/mL and 137.4 μg/mL). In addition, we also performed in vitro cytotoxic assays toward peritoneal macrophages and J774 macrophages. For the in vitro cytotoxicity assay employing J774 cells, all of the sulfated polysaccharides decreased cell survival, with CC₅₀ values of 27.3 μg/mL, 49.3 μg/mL, 73.2 μg/mL, and 99.8 μg/mL for Bo, Cr, Gc, and Sf, respectively. However, none of the sulfated polysaccharides reduced the cell growth rate of the peritoneal macrophages. These results suggest that macroalgae contain compounds with various chemical properties that can control specific pathogens. According to our results, the assayed sulfated polysaccharides were able to modulate the growth rate and cell survival of Leishmania (L.) amazonensis promastigotes in in vitro assays, and these effects involved the interaction of the sulfated polysaccharides on the cell membrane of the parasites.     

膜过滤绿茶多糖的系统分级纯化及免疫活性分析

采用150kD、20kD、6kD的超滤膜和对水溶性茶多糖进行分级和浓缩,得到的三部分截留液分别经过DEAE-52纤维素离子交换层析和丙烯葡聚糖凝胶Sephacryl S-300柱层析系统分离、纯化,共得到了20多个分级组分;HPGPC-ELSD鉴定各多糖组分的纯度及分子量分布,结果表明得到5个均一多糖组分;用小鼠巨噬细胞系Raw264.7检测免疫活性,发现对小鼠巨噬细胞Raw264.7有显著活性的多糖组分大多集中在20kD左右,大多为不均一多糖。     

Ascophyllan Purified from Ascophyllum nodosum Induces Th1 and Tc1 Immune Responses by Promoting Dendritic Cell Maturation

Marine-derived sulfated polysaccharides have been shown to possess certain anti-virus, anti-tumor, anti-inflammatory and anti-coagulant activities. However, the in vivo immunomodulatory effects of marine-derived pure compounds have been less well characterized. In this study, we investigated the effect of ascophyllan, a sulfated polysaccharide purified from Ascophyllum nodosum, on the maturation of mouse dendritic cells (DCs) in vitro and in vivo. Ascophyllan induced up-regulation of co-stimulatory molecules and production of pro-inflammatory cytokines in bone marrow-derived DCs (BMDCs). Moreover, in vivo administration of ascophyllan promotes up-regulation of CD40, CD80, CD86, MHC class I and MHC class II and production of IL-6, IL-12 and TNF-α in spleen cDCs. Interestingly, ascophyllan induced a higher degree of co-stimulatory molecule up-regulation and pro-inflammatory cytokine production than fucoidan, a marine-derived polysaccharide with well-defined effect for promoting DC maturation. Ascophyllan also promoted the generation of IFN-γ-producing Th1 and Tc1 cells in the presence of DCs in an IL-12-dependent manner. Finally, myeloid differentiation primary response 88 (MyD88) signaling pathway was essential for DC maturation induced by ascophyllan. Taken together, these results demonstrate that ascophyllan induces DC maturation, and consequently enhances Th1 and Tc1 responses in vivo. This knowledge could facilitate the development of novel therapeutic strategies to combat infectious diseases and cancer.     

酶法修饰绿茶多糖对免疫低下模型小鼠免疫活性的影响

以糖苷酶溶液为工具酶,研究酶法修饰茶多糖对环磷酰胺抑制的免疫低下模型小鼠的免疫功能的影响。结果表明,原多糖TPS与改性后的茶多糖ETPS在免疫低下模型小鼠的细胞免疫功能、体液免疫功能、单核-巨噬细胞功能和NK细胞活性4个方面的实验中,结果均呈阳性,说明它们均具有明显的增强免疫功能的作用,酶法修饰改性能显著提高绿茶多糖的免疫活性。在同等剂量下,ETPS与TPS相比能提高脾指数、腹腔巨噬细胞吞噬率和吞噬指数,能显著提高DTH试验耳肿程度、血清HC50、NK细胞活性。绿茶多糖通过酶法修饰免疫活性得到增强。     

Salt Effect on the Antioxidant Activity of Red Microalgal Sulfated Polysaccharides in Soy-Bean Formula

Sulfated polysaccharides produced by microalgae, which are known to exhibit various biological activities, may potentially serve as natural antioxidant sources. To date, only a few studies have examined the antioxidant bioactivity of red microalgal polysaccharides. In this research, the effect of different salts on the antioxidant activities of two red microalgal sulfated polysaccharides derived from Porphyridium sp. and Porphyridium aerugineum were studied in a soy bean-based infant milk formula. Salt composition and concentration were both shown to affect the polysaccharides’ antioxidant activity. It can be postulated that the salt ions intefer with the polysaccharide chains’ interactions and alter their structure, leading to a new three-dimensional structure that better exposes antiooxidant sites in comparison to the polysaccharide without salt supplement. Among the cations that were studied, Ca²⁺ had the strongest enhancement effect on antioxidant activities of both polysaccharides. Understanding the effect of salts on polysaccharides’ stucture, in addition to furthering knowledge on polysaccharide bioactivities, may also shed light on the position of the antioxidant active sites.     

Structural Analysis and Anti-Complement Activity of Polysaccharides from Kjellmaniella crsaaifolia

Two polysaccharides, named KCA and KCW, were extracted from Kjellmaniella crassifolia using dilute hydrochloric acid and water, respectively. Composition analysis showed that these polysaccharides predominantly consisted of fucose, with galactose, mannose and glucuronic acid as minor components. After degradation and partial desulfation, electrospray ionization mass spectrometry (ESI-MS) was performed, which showed that the polysaccharides consisted of sulfated fucooligosaccharides, sulfated galactofucooligosaccharides and methyl glycosides of mono-sulfated/multi-sulfated fucooligosaccharides. The structures of the oligomeric fragments were further characterized by electrospray ionization collision-induced dissociation tandem mass spectrometry (ESI-CID-MS² and ESI-CID-MS³). Moreover, the activity of KCA and KCW against the hemolytic activity of both the classical and alternative complement pathways was determined. The activity of KCA was found to be similar to KCW, suggesting that the method of extraction did not influence the activity. In addition, the degraded polysaccharides (DKCA and DKCW) displayed lower activity levels than the crude polysaccharides (KCA and KCW), indicating that molecular weight had an effect on activity. Moreover, the desulfated fractions (ds-DKCA and ds-DKCW) showed less or no activity, which confirmed that sulfate was important for activity. In conclusion, polysaccharides from K. crassifolia may be good candidates for the treatment of diseases involving the complement pathway.     

抗菌卷柏多糖的提取与分离

采用4种方法提取卷柏多糖,其中综合法提取率最高,为1．45％。可溶性多糖经DEAE-纤维素52柱层析进一步纯化,得到1种中性糖和1种酸性糖,各占86．5％和13．5％。利用纸片扩散法测定其抗菌活性,结果表明,卷柏多糖具有较强的抗动植物病原菌活性,对肠炎病病原菌、绿木霉与甘薯薯瘟病病原菌的最小抑菌浓度为6．25mg·mL^-1。     

The Application of Seaweed Polysaccharides and Their Derived Products with Potential for the Treatment of Alzheimer’s Disease

Neurodegenerative diseases are among the most widespread diseases affecting humans, and the number of patients is only rising. Seaweed polysaccharide extracts show significant neuroprotective and reparative activities. Seaweed polysaccharides might provide the next big breakthrough in neurodegenerative disease treatment. This paper reviews the applications of seaweed polysaccharides as potential treatments of neurodegenerative diseases. The particular focus is on fucoidan, ulvan, and their derivatives as potential agents to treat Alzheimer’s disease. This review provides a critical update on the progress in this important research area.     

Extraction Procedure,Characteristics, and Feasibility of Caulerpa microphysa (Chlorophyta) Polysaccharide Extract as a Cosmetic Ingredient

The green alga Caulerpa microphysa, which is native to Taiwan, has a relatively high economic value and a well-developed culture technique, and is used mainly as a foodstuff. Its extract has been shown to exhibit antitumor properties, but the polysaccharide content of the extract and its anti-inflammatory and wound-healing effects and moisture-absorption and -retention capacity remain unknown. Hence, the objective of this study was to evaluate the potential of the polysaccharides in C. microphysa extract (CME) for use in cosmetics. The overall polysaccharide yield from the CME was 73.93% w/w, with four molecular weight fractions. The polysaccharides comprised 59.36 mol% mannose, 27.16 mol% glucose, and 13.48 mol% galactose. In addition, the CME exhibited strong antiallergic, wound-healing, transdermal-delivery, and moisture-absorption and -retention effects. In conclusion, the results suggested that CME potentially has anti-inflammatory and wound-healing effects and a good moisture capacity, which can be used in cosmetic applications.     

乌龙茶多糖调节血脂作用及其机制研究

探讨了乌龙茶多糖（OTPS）对高脂模型小鼠血脂的调节作用及机制。采用高脂饲料饲喂法建立高脂血症小鼠模型,通过不同剂量OTPS灌胃6周后,测定了小鼠的体重、胸腺重量、脾脏重量、肝脏重量、血脂水平、血清和肝脏中丙二醛（MDA）和超氧化物歧化酶（SOD）的含量,以及观察了肝脏的病理变化。结果表明,与高脂模型对照组比较,OTPS剂量组血清总胆固醇（TC）和低密度脂蛋白（LDL-C）、血清和肝脏MDA、肝脏指数显示不同程度的降低,胸腺指数、脾指数、血清和肝脏SOD活力则有所上升,肝脏病变有一定程度的减轻,说明乌龙茶多糖具有一定的调节血脂、提高机体免疫力及抗氧化的作用。     

茶多酚与多糖的相互作用:作用机理及功能特性变化研究进展

茶多酚具有抗氧化、抗肿瘤、抗辐射等多种生物活性,与环糊精、黑木耳多糖或壳聚糖等多种多糖作用后,功能活性增强,但相互作用的过程与机理尚不明确。因此,对茶多酚与多糖的结构及其相互作用的研究显得尤为重要。近几年,关于茶多酚与多糖之间相互作用的研究日益增多,本文介绍了茶多酚和多糖的分类及结构,概括了多糖与茶多酚之间的相互作用类型、影响因素及研究方法,针对二者之间的相互作用对茶多酚功能特性的影响进行了综述,为后续研究二者与其他活性物质相互作用以及功能特性变化等方面提供参考。     

Antihyperglycemic, Antihyperlipidemic and Antiglycation Effects of Byrsonima crassifolia Fruit and Seed in Normal and Streptozotocin-Induced Diabetic Rats

The hypoglycemic effects of hexane, chloroform and methanol extracts from fruits and seeds of Byrsonima crassifolia were evaluated by oral administration to normoglycemic and streptozotocin-induced severe diabetic rats (SD). The anti-diabetic effect was examined by blood glucose, triglycerides, lipid peroxidation, total cholesterol levels in the serum, glycogen content of liver and skeletal muscles, superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and oxidized glutathione (GSSG) levels. The most active extracts were obtained with hexane. Hexane and chloroform extracts from fruits and seeds of Byrsonima crassifolia increased the levels of SOD, GSH, GSSG and CAT, hepatic glycogen content, glucose-6-phosphatase (G6Pase) and the plasma insulin levels. They also decreased glucokinase (GK) and TBAR (thiobarbituric acid assay). In conclusion, Byrsonima crassifolia possesses significant antihyperglycemic properties after 4 h of a single oral dose. It can also improve hyperlipidemia and hyperinsulinemia in streptozotocin-induced diabetic rats. Both extracts exhibited significant inhibitory activity against AGEs (advanced glycation end products) formation with IC₅₀ values ranging from 94.3 to 138.7 μg/ml. Therefore, B. crassifolia can be considered as a potential safe anti-diabetic agent.     

三七丹参片功能学实验研究

目的通过经口给予小鼠不同剂量三七丹参片实验,验证三七丹参片的增强免疫力功能,为临床使用提供依据。方法实验设低、中、高三个剂量组,分别相当于人体推荐摄入量的5、10、30倍,同时设阴性对照组(1%羧甲基纤维素钠),连续一个月经口给予小鼠不同剂量的受试物。结果与阴性对照组比较,三个剂量组小鼠血清溶血素抗体积数值均升高;中、高剂量组小鼠腹腔巨噬细胞吞噬鸡红细胞的吞噬率、吞噬指数均升高;以上差异均具有显著性意义(P<0.05,P<0.01)。结论三七丹参片具有增强免疫力功能。     

Effect of dietary rosemary extract on cell-mediated immunity of young rats

The impact of rosemary extract on splenic mononuclear cell proliferation was determined. Weanling male Sprague-Dawley rats were fed diets containing 0, 100, 200 or 400 ppm rosemary extract or 400 ppm butylated hydroxytoluene (BHT) in combinatiion with 10 or 20% casein enriched diets for 8 weeks. Splenic mononuclear cells were isolated from these animals and mitogenic response to Concanavalin A (Con A), Phytohemagglutinin (PHA) and lipopolysaccharide was determined. Con A and PHA-stimulated proliferation of spleen cells from rats fed 10% casein and 200 ppm rosemary extract was significantly higher than that of cells from the corresponding control animals. However, other levels of rosemary at 10% dietary casein or rosemary at any concentration fed along with 20% dietary casein had no impact on the mitogenic stimulation of splenic mononuclear cells. Thus, these results suggest that the use of rosemary might not have a generalized immunoenhancing effect, and will probably be effective in some stressed conditions, such as protein or antioxidant deficiency.     

Altered expression of circadian clock gene, mPer1, in mouse brain and kidney under morphine dependence and withdrawal

Every physiological function in the human body exhibits some form of circadian rhythmicity. Under pathological conditions, however, circadian rhythmicity may be dusrupted. Patients infected with HIV or addicted to drugs of abuse often suffer from sleep disorders and altered circadian rhythms. Early studies in Drosophila suggested that drug seeking behavior might be related to the expression of certain circadian clock genes. Our previous research showed that conditioned place preference with morphine treatment was altered in mice lacking the Period-1 (mPer1) circadian clock gene. Thus, we sought to investigate whether morphine treatment could alter the expression of mPer1, especially in brain regions outside the SCN and in peripheral tissues. Our results using Western blot analysis showed that the mPER1 immunoreactivity exhibited a strong circadian rhythm in the brains of the control (Con), morphine-dependent (MD), and morphine-withdrawal (MW) mice. However, the phase of the circadian rhythm of mPER1 expression in the brains of MD mice significantly differed from that of the Con mice (p < 0.05). In contrast to mPER1 expression in the brain, the circadian rhythm of mPER1 immunoreactivity in the kidneys was abolished after morphine administration, whereas the Con mice maintained robust circadian rhythmicity of mPER1 in the kidney. Therefore, the effect of morphine on the circadian clock gene mPer1 may vary among different organs, resulting in desynchronization of circadian function between the SCN and peripheral organs.     

茶多糖和茶多酚的降血糖作用研究

目的:研究茶多糖、茶多酚对四氧嘧啶致糖尿病SD大鼠的降血糖作用和机制。方法:饲喂SD大鼠茶多糖、茶多酚3周后,观察大鼠血糖、葡萄糖耐量、血胰岛素以及小肠糖降解酶(淀粉酶、蔗糖酶、麦芽糖酶)变化。结果:茶多糖、茶多酚都有显著抑制糖尿病大鼠血糖升高的作用;与对照组比较,茶多糖组大鼠血胰岛素水平有显著提高(P<0.05),蔗糖酶和麦芽糖酶活性显著降低(P<0.05);茶多酚组的血胰岛素水平有升高趋势,小肠各降解酶活力也有下降趋势,但与对照组比较均未达到显著水平。结论:茶多糖对高血糖大鼠有显著的抑制血糖升高的作用,茶多糖的作用机制可能是抑制小肠糖降解酶活性。     

中低档绿茶中茶多糖的提取及降血糖作用

从中低档绿茶中提取茶多糖，分离提取和精制纯化的方法对其含量及生理活性有一定影响。理化处理和分析结果表明，茶多糖是一类与蛋白质结合在一起的酸性多糖或一种酸性糖蛋白，以丙酮为溶剂沉淀分离得到的茶多糖生理活性最高，降血糖作用最佳，而且方法操作简便，易于工业化生产。茶多糖对正常和实验性高血糖模型小白鼠的血糖均有显著的降低作用。茶多糖在降低四氧嘧啶高血糖小鼠血糖浓度的同时，使肝糖原大量增加，表明茶多糖在对糖代谢方面具有与胰岛素相类似的作用。     

Polysaccharide from Codium fragile Induces Anti-Cancer Immunity by Activating Natural Killer Cells

Natural polysaccharides exhibit beneficial immune modulatory effects, including immune stimulatory and anti-cancer activities. In this study, we examined the effect of Codium fragile polysaccharide (CFP) on natural killer (NK) cell activation, and its effect on tumor-bearing mice. Intravenous CFP treatment of C57BL/6 mice resulted in the upregulation of CD69, which is a marker associated with NK cell activation. In addition, intracellular levels of interferon (IFN)-γ and the cytotoxic mediators perforin and granzyme B were markedly increased in response to the CFP treatment of splenic NK cells. IFN-γ production by NK cells was directly induced by CFP, whereas the upregulation of CD69 and cytotoxic mediators required IL-12. Finally, intraperitoneal treatment with CFP prevented CT-26 (murine carcinoma) tumor cell infiltration in the lungs, without significantly reducing the body weight. In addition, treatment with CFP prevented B16 melanoma cell infiltration in the lung of C57BL/6 mice. Moreover, the anti-tumor effect was diminished by the depletion of NK cells. Therefore, these data suggest that CFP may be used as an NK cell stimulator to produce a phenomenon that contributes to anti-cancer immunity.