柳生金针菇胞外粗多糖对小鼠肝癌H22移植性肿瘤的抑制作用和免疫功能调节的研究

本研究通过对柳生金针菇胞外粗多糖对肿瘤生长的抑制和机体免疫调节的体内实验,来探索柳生金针菇胞外粗多糖的相关抗肿瘤机制。结果表明柳生金针菇胞外粗多糖高、中、低剂量组能不同程度抑制肝癌H22肿瘤的生长。高剂量组抑瘤率最高,为35.25%,中低剂量也能抑制肿瘤生长,缓解脾脏肿大和胸腺萎缩。同时免疫学实验结果表明柳生金针菇胞外粗多糖可明显增加血清中各种细胞因子的表达,通过增强细胞和体液免疫功能来起到抗肿瘤作用。     

深层发酵香菇水溶性胞外多糖的生物学活性

由香菇ＣＬ－２菌丝发酵上清液中分离到水溶性胞外多糖（ＨＥＰ）。研究表明,ＨＥＰ具有较强的免疫增效作用。它能有效促进正常小鼠腹腔巨噬细胞的吞噬功能,显著提高Ｔ淋巴细胞的百分含量,对小鼠体液免疫也有促进作用。ＨＥＰ对肉瘤Ｓ１８０的抑制率达３９．７％,并能显著延长荷瘤（ＥＡＣ）小鼠的存活时间,延长率达４０．５％。ＨＥＰ对牛艾滋病毒（ＢＩＶ）有直接抑制作用,抑制率为６６．７％。     

紫球藻及其胞外多糖对小鼠免疫功能的调节作用

探讨紫球藻及其胞外多糖对免疫低下小鼠免疫功能的调节作用,用环磷酰胺(CY)建立昆明种小鼠免疫功能低下的实验模型。小鼠随机均分6组,其中2组每天分别给予紫球藻藻粉1200、600 mg/kg灌胃,2组给予紫球藻胞外多糖300、150 mg/kg灌胃,正常对照组及CY组用等容积生理盐水灌胃,连续14 d。实验结果表明紫球藻及其胞外多糖均可显著提高免疫抑制小鼠的脾指数、胸腺指数、碳廓清能力、单核细胞吞噬功能,并可对抗环磷酰胺引起的外周血白细胞数目下降,因此紫球藻藻粉及其胞外多糖对小鼠免疫功能具有一定的正向调节作用。毒性实验表明,小鼠腹腔注射300、150 mg/kg紫球藻胞外多糖溶液和分别给予紫球藻藻粉1200、600 mg/kg灌胃,小鼠未出现死亡,也未有明显毒性反应出现的一些指标变化,说明紫球藻藻粉及胞外多糖的安全性较高。     

细虫草胞外多糖对小鼠腹腔巨噬细胞免疫功能研究

本实验在体外条件下,以人工发酵培养的细虫草胞外多糖OgE、OgE-F1和OgE-F2作用于小鼠腹腔巨噬细胞RAW264.7,通过测定其对巨噬细胞的增殖率、代谢MTT活力、NO分泌和吞噬能力的影响,评价细虫草胞外多糖的免疫调节活性。结果表明,细虫草多糖对巨噬细胞无细胞毒性,且能促进巨噬细胞代谢MTT活力;在0.2mg/mL^1.0mg/mL浓度范围内,多糖呈剂量依赖性的促进巨噬细胞分泌NO水平和吞噬能力。本研究表明,细虫草多糖能有效地增强小鼠巨噬细胞的活性,潜在地可改善小鼠的先天性免疫调节。     

啤酒酵母胞外多糖发酵条件的研究

以啤酒酵母S－12为出发菌株,用紫外线＋氯化锂作为复合诱变剂,获得一株产胞外多糖量较高的变株S－12－4,比出发菌株提高33．3％,同时对变株进行了最佳培养条件的研究,结果表明：最适碳源和氮源分别为大米糖3％、酵母粉0．37％及NH4Cl0．32％,最适发酵条件为起始PH6．0、培养温度26℃,发酵周期为30h,在此基础上进行培养,变株S－12－4产胞外多糖最高可达38.2mg/100mL,比初始条件提高了54％。     

深层发酵香菇水溶性胞外多糖的生物学活性

由香菇（Lentinusedodes）CL-2菌丝发酵上清液中分离到水溶性胞外多糖（HEP）。研究表明,HEP具有较强的免疫增效作用。它能有效促进正常小鼠腹腔巨噬细胞的吞噬功能,显著提高T淋巴细胞的百分含量,对小鼠体液免疫也有促进作用。HEP对肉瘤S(180)的抑制率达39.7％,并能显著延长荷瘤（EAC）小鼠的存活时间,延长率达40.5％。HEP对牛艾滋病毒（BV）有直接抑制作用,抑制率为66.7％。     

假单胞菌胞外多糖发酵条件的研究

研究了假单胞菌（Ｐｓｅｕｄｏｍｏｎａｓｓｐ．６）利用木糖产胞外多糖的发酵条件。实验表明ＫＨ２ＰＯ４,Ｏ２和高碳氮比对多糖合成有促进作用,在发酵后期补加木糖有助于多糖产量的提高     

一株乳酸菌所产胞外多糖对荷瘤小鼠机体免疫功能影响的研究

对一株产胞外多糖（Exopolysaccharide,简称EPS）的乳酸菌Z222菌株的培养条件进行了优化, 其发酵液通过浓缩、除蛋白、脱色、乙醇沉淀、透析、CM纤维素柱、DEAESephadex 离子交换柱和Sephadex G100凝胶柱层析,冷冻干燥后得到一种纤维状白色固体产品EPSⅠ。将该产品对荷肉瘤S180小鼠进行机体免疫功能影响的初步试验,统计学处理结果表明,它对迟发型超敏反应、免疫器官重量和脾细胞抗体形成均有较为明显的影响。EPSⅠ有希望作为一种新的免疫调节剂。     

粗柄羊肚菌菌丝体多糖及胞外多糖的提取工艺

为了研究粗柄羊肚菌菌丝体多糖及胞外多糖的最佳提取工艺条件,采用超声波提取菌丝体多糖、菌丝体发酵液浓缩提取胞外多糖的方法。通过单因素试验和L9(34)正交试验设计,探讨二者的最佳提取工艺条件。研究结果表明菌丝体多糖的最佳提取工艺条件:料液比1∶20(g∶m L),超声波处理温度70℃,超声时间20 min,超声提取次数2次;胞外多糖的最佳提取工艺条件:浓缩倍数1∶5,浓缩温度50℃,醇沉浓度95%,p H为6。该项工艺研究得到菌丝体粗多糖平均含量56.761 2 mg/g,胞外多糖平均含量1.275 4 mg/m L,多糖产量稳定,且此试验方法稳定可行。     

Immunomodulating activities of water-soluble exopolysaccharides obtained from submerged culture of Lentinus lepideus

Immunomodulating activities of water-soluble exopolysaccharides (LL-EX) obtained from submerged mycelial culture of Lentinus lepideus were studied and their effectiveness was compared with lipopolysaccharide (LPS). The influence of the LL-EX on macrophage cellular lysosomal enzyme activity was to stimulate up to 267%, 392%, and 464% at the level of 10, 50, and 100 mug/ml, respectively. When the LL-EX was further fractionated into LL-Fr.I and Fr.II by Sepharose CL-6B gel chromatography, the cellular lysosomal enzyme activity of LL-Fr.II (2.1- fold) was higher than Fr.I (1.2-fold). Moreover, both LL-Fr.I and Fr.II stimulated the cytokines IL-1beta, TNF-alpha, and IL-6 in macrophages. In mixed lymphocyte reaction, LL-Fr.I and Fr.II enhanced the splenocyte proliferation up to 1.2-fold and 1.4-fold (50 mg/ml), respectively, stimulating only T lymphocytes. The fractions of LL-EX not show any direct toxicity against human gastric adenocarcinoma cell (AGS). The molecular masses of LL-Fr.I and Fr.II were estimated to be about 1,986 kDa and 21 kDa, respectively. The total sugar and protein contents of the two fractions were 84.97% and 69.88% and 15.03% and 30.12%, respectively. The sugar and amino acid compositions of the LL-Fr.I and Fr.II were also analyzed in detail.     

参芝发酵产物的抗肿瘤活性及其对小鼠免疫功能的影响

为了探索红参水煎液的灵芝发酵产物对H22荷瘤小鼠的抗肿瘤活性及其对小鼠免疫功能的影响,通过体内抗肿瘤实验和增强免疫功能实验从抑瘤率、对免疫器官的影响指数、对非特异性免疫、体液免疫及细胞免疫的影响5个方面对该产物做了功能性评价。结果表明,在抗肿瘤实验中,参芝发酵产物高剂量组的抑瘤率达到51.65%,脾指数和胸腺指数均高于对照组和环磷酰胺(CTX)组;增强免疫功能实验中,3个实验的给药组小鼠和对照组小鼠相比都有显著性差异(P<0.01)。由此可见,将灵芝与人参在发酵层次上配伍具有显著的抑制荷瘤小鼠肿瘤生长及增强小鼠免疫功能的作用。     

红缘拟层孔菌固体发酵产物体内抗肿瘤和抗氧化作用研究

研究了红缘拟层孔菌固体发酵产物对H22荷瘤小鼠的抗肿瘤和抗氧化作用,以抑瘤率、脾和胸腺指数、白细胞介素-2、干扰素-r、血管内皮细胞生长因子、超氧化物歧化酶、丙二醛、过氧化氢酶、谷胱甘肽过氧化物等指标来考察红缘拟层孔菌固体发酵产物对H22荷瘤小鼠肿瘤抑制和体内抗氧化作用。结果表明,固体发酵产物高剂量和中剂量组的抑瘤率分别为66.66%和64.70%,与阴性组比较,有显著的抗肿瘤作用（P<0.01）,HE染色切片也能观察到固体发酵产物高、中和低各组肿瘤细胞大量坏死,并且高剂量和低剂量组血清中白细胞介素-2和干扰素-r含量显著增加,血管内皮生长因子含量降低,与抑制肿瘤效果具有一定相关性;此外,红缘拟层孔菌固体发酵产物各组可以降低丙二醛含量,提高超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物含量;综上所述,红缘拟层孔菌固体发酵产物具有显著的抗肿瘤和抗氧化作用。     

Production and isolation of chitosan by submerged and solid-state fermentation from Lentinus edodes

A method for the laboratory-scale production and isolation of chitosan (polyglucosamine) by liquid and solidstate fermentation from Lentinus edodes was developed. The yields of isolated chitosan were 120 mg/L of fermentation medium under liquid fermentation conditions and 6.18 g/kg of fermentation medium under solid-state fermentation conditions. The latter method, which gives up to 50 times yields than other chitosan production methods from fungi, provides a new flexible and easily scaledup procedure for the production of low acetylation degree chitosan. (c) 1996 John Wiley & Sons, Inc.     

Factors influencing the production of water-soluble endopolysaccharides and exopolysaccharides from Lentinus lepideus and their effects on immune cytokine production

An efficient method to produce water-soluble polysaccharides from Lentinus lepideus is described. The productivity of both endopolysaccharides (PPS) and exopolysaccharides (EPS) was compared under various culture conditions. The effect of treating their own PPS and EPS on immune cytokine production was also studied in relation to culture factors. High yield production of EPS required moderate culture temperature (25 degrees ) as well as long culture period (16-20 days). In contrast, PPS production required high culture temperature (30 degrees ) and short culture period ( days). Most of the carbon sources did not affect polysaccharides and mycelial production except for sucrose. Immune cytokine levels in the EPS treatment varied among carbon sources or culture periods. PPS did not appear to affect much on the production of cytokines, regardless of the culturing factors, except for the culture period. These results suggest that the optimal culture conditions for L. lepideus vary according to culture purposes, and different culture conditions should be used for different targets including mycelial biomass, EPS, and PPS. Whereas the immunomodulating activity of EPS appeared to be affected by culture conditions in L. lepideus, that of PPS did not.     

Intermediate chains of exopolysaccharides from Lactobacillus rhamnosus RW‐9595M increase IL‐10 production by macrophages

Aims: To evaluate the immunosuppressive properties of the exopolysaccharide (EPS) from high‐EPS producer Lactobacillus rhamnosus RW‐9595M on inflammatory cytokines produced by macrophages. Methods and Results: The conditioned media (CM) were produced by macrophages treated with parental Lact. rhamnosus ATCC 9595 and its isogenic variant, the high‐EPS producer Lact. rhamnosus RW‐9595M, and the levels of TNF‐α, IL‐6, IL‐10 and IL‐12 were evaluated. Results revealed that CM from parental Lact. rhamnosus induced higher levels of TNF‐α, IL‐6 and IL‐12 but inhibited IL‐10 production, whereas its mucous variant induced low or no TNF‐α and IL‐6. Addition of purified EPS to macrophages treated with parental Lact. rhamnosus decreased the inflammatory cytokines and inhibited the metabolic activity of lymphocytes. The intermediate polysaccharide chains (16–30 units) produced by time‐controlled hydrolysis of EPS increased the IL‐10 produced by macrophages. Conclusions: Polysaccharide chains of EPS induced immunosuppression by the production of macrophagic anti‐inflammatory IL‐10. Significance and impact of the Study: These results indicate that the EPS from Lact. rhamnosus RW‐9595M may be useful as a new immunosuppressive product in dairy food.     

响应面分析法优化黑根霉胞外多糖发酵工艺

实验以商品化的马铃薯葡萄糖液体培养基为基础培养基,以胞外粗多糖产量为考察指标,运用响应面分析法考察玉米浆浓度、KH₂PO₄浓度和发酵时间3个因素对胞外多糖发酵产量的影响,以获得黑根霉胞外多糖发酵最优工艺,建立高产、稳定、可控的胞外多糖发酵生产工艺技术方案。经响应面分析,各因素按照对响应值的影响顺序为：玉米浆浓度>发酵时间> KH₂PO₄浓度,且玉米浆浓度、发酵时间对胞外多糖产量的影响极显著,KH₂PO₄浓度对胞外多糖产量的影响不显著。胞外多糖发酵最优工艺为：玉米浆3.2mg/mL、KH₂PO₄ 1.5mg/mL和发酵时间132h,在此条件下胞外多糖的最大预测产量为0.824mg/mL。实验重复性好,是一个高产、稳定、可控的胞外多糖发酵生产工艺技术方案,可以指导黑根霉胞外多糖发酵。     

香菇菌丝体多糖的分离鉴定与免疫功能研究

从香菇（Ｌｅｎｔｉｎｕｓｅｄｏｄｅｓ）菌丝体提取得到的粗多糖,经ＤＥ５２柱层析,得到均一的多糖成分,命名为ＬＥ。其平均分子量为５．０８×１０５,ＬＥ经红外（ＩＲ）和紫外（ＵＶ）光谱分析,为多糖蛋白质复合体,其中糖含量为９４．２％,蛋白质含量为５．８％。完全酸水解表明糖组成为单一的葡萄糖,蛋白质组成主要为Ａｌａ等１６种天然氨基酸。ＬＥ经完全甲基化、酸水解、乙酰化、薄层层析（ＴＬＣ）和［１Ｈ］核磁共振（［１Ｈ］ＮＭＲ）分析,确定其多糖部分的基本结构为１→３连接的葡聚糖,含有部分１→６侧链。此外,用反转录聚合酶链式反应（ＲＴＰＣＲ）和生物测定法分别研究了健康人外周血单核细胞中白细胞介素２（ＩＬ２）和肿瘤坏死因子（ＴＮＦα）的基因表达和活性,结果发现ＩＬ２和ＴＮＦα的基因表达和活性均高于空白对照组,这表明ＬＥ可诱导某些免疫反应。