灵芝孢子粉破壁技术、质量分析与深加工相关研究进展

破壁灵芝孢子粉为一种灵芝深加工产品,具有多种药理学活性,更易被人体吸收。灵芝孢子的破壁方法包括生物化学法、物理法和机械法等,工业生产多综合利用多种方法。破壁灵芝孢子粉的质量分析还没有统一的国家标准,灵芝孢子粉质量可以参考地方标准进行综合评价,还可以参考近期发表的一些分析方法。灵芝孢子粉提取物产品以孢子油为主,其它产品也有研发并推出市场。药理学研究表明,灵芝孢子油、灵芝孢子粉水提物和醇提物均具有抗癌活性,有良好的应用前景。     

RP-HPLC法测定不同部位灵芝中灵芝酸B含量

目的：测定灵芝中具有苦味的成分灵芝酸B在灵芝中不同部们的含量,方法：采用RP－HPLC法测定灵芝菌盖的表皮层,木栓层,菌柄,孢子粉,菌盖子实体等部位分中灵芝酸B的含量,结果：灵芝药材中灵芝酸B主要集中在灵芝的表皮部位,其它部位则较少,结论：为寻找灵过的有效成分分布规律提供了客观依据。     

不同生长阶段灵芝中微量元素与重金属含量分析

利用ICP-MS、AFS和AAS对同一菌种灵芝深层发酵菌丝体、不同生长阶段的子实体及孢子粉中微量元素与重金属含量进行分析。发酵菌丝体、子实体及孢子粉中均含有生物必需微量元素和重金属,以孢子粉中含量相对较高。硒的含量在各个阶段几乎相同,而锗的含量则呈现生物富集的现象。     

一种灵芝孢子粉收集器的研究

目的:开发一种方便实用、低本高效的灵芝孢子粉收集器,收集优质灵芝孢子粉.方法:利用白卡纸或油光纸等通用材料,生产灵芝孢子粉收集器,以温度、湿度为主要指标比较孢子粉采集器对灵芝生长环境的影响,并调查集粉效果.结果:研发的灵芝孢子粉采集器能有效地把灵芝子实体与外界相对隔离,形成一个基本封闭的独立空间,阻止了孢子粉的自由飘散;孢子粉采集器的使用对灵芝生长的温度与湿度等环境影响小,不仅不会影响灵芝子实体发育,而且因采集器内空气相对湿度变化更趋平缓而更有利于孢子粉的散发.结论:该灵芝孢子粉采集器孢子粉收集效率高、质量好,大面积应用时灵芝孢子粉产量可达孢子粉与灵芝子实体总量的38.3%.     

灵芝菌丝营养生长特征与子实体形成相关性研究

目的:选择适于代料栽培的灵芝菌种。方法:利用培养皿及三角瓶进行固体及液体培养,后期以瓶栽结果验证。结果:易出多孔组织的菌株瓶栽时子实体形成快,孢子产量高。结论:菌丝阶段出现多孔组织的难易可作为代料栽培菌种选择的特性与依据。     

不同品种灵芝中三萜类化合物的比较研究

本研究利用HPLC对同一灵芝菌种不同生长阶段及不同品种灵芝子实体中三萜类化合物的含量和图谱进行了分析.结果表明在液体发酵7天左右的灵芝菌丝体中灵芝三萜类化合物的含量极低,而不同生长阶段的灵芝子实体中的三萜类化合物的变化较小,孢子粉中三萜类化合物的含量较子实体中低.不同品种的灵芝子实体中三萜类化合物的种类和含量存在差异,黑芝中几乎不含三萜类化合物.     

赤芝孢子粉和子实体主要化学成分变异规律研究

目的:揭示产地、品种、采收部位和潮次、破壁技术对赤芝孢子粉和子实体中总三萜、总多糖、总蛋白含量的影响。方法:收集龙芝1号、沪农1号、市售赤芝孢子粉和子实体,分别采用硫酸-蒽酮比色法、香草醛-冰醋酸比色法和Lowry法测定其中总多糖、总三萜、总蛋白的含量。结果:全国6个主产区的破壁孢子粉总多糖为0.40%~2.25%,总三萜为1.36%~3.15%,总蛋白为0.74%~1.91%;浙江主栽赤芝品种子实体总多糖、总三萜、总蛋白质量分数分别为0.25%~1.42%,0.44%~1.42%,1.82%~3.67%,未破壁孢子粉分别为0.41%~0.91%,0.09%~0.12%,0.78%~0.90%,破壁孢子粉分别为1.03~2.25%,1.89%~3.15%,0.96%~1.04%。结论:全国不同主产区的灵芝破壁孢子粉化学成分含量存在显著差异,浙江产区总多糖和总三萜含量较高,福建产区总蛋白含量较高;浙江主栽赤芝品种龙芝1号总三萜含量较高,沪农1号总多糖含量较高;破壁孢子粉中总三萜与总多糖含量高于子实体,子实体菌柄中总多糖含量大多高于菌盖,菌盖中总三萜含量显著高于菌柄;2潮采收的菌盖和菌柄中总多糖含量高于1潮相应的部位,但总三萜含量大多低于1潮;孢子粉破壁可显著提高总三萜和总多糖的提取和溶出率。     

灵芝提取物对人外周血细胞免疫调节作用的研究

 目的 研究灵芝子实体的水提取物、醇提取物单独及与诱导物一起使用对人外周血吞噬细胞和淋巴细胞的免疫调解作用。方法 以吞噬细胞化学发光、淋巴细胞化学发光和淋巴细胞增殖的方法,观察灵芝提取物的免疫调节作用。结果 单独灵芝提取物不能引起细胞免疫反应,与诱导物一起使用后,低浓度的灵芝提取物对细胞免疫功能有激活作用,高浓度则有免疫抑制作用。结论 与诱导物一起使用后,灵芝水提取物和醇提取物对人外周血吞噬细胞和淋巴细胞均具有双向调节作用。     

Inhibitive effect on apoptosis in splenic lymphocytes of mice pretreated with Lingzhi (Ganoderma Lucidum) spores

Objective

To investigate how the pretreatment of mice with Ganoderma spores affected the apoptosis of their splenic lymphocytes induced by dexamethasone after 19 days treatment.

Methods

Sixty Kunming mice were randomly divided into six groups: blank control groupdrenched with normal saline; a drug control group drenched with 150 mg/mL Ganoderma spores; a model group treated with saline; a low dose group with 50 mg/mL Ganoderma spores; a moderate dose group with 100 mg/mL Ganoderma spores; and a high dose group with 150 mg/mL Ganoderma spores. The effect of Ganoderma spores on apoptosis in spleen lymphocytes was analyzed. All groups were treated for 19 days. On day 20, the model group and the 3 treatment groups were intraperitoneally injected dexamethasone to induce apoptosis. Splenic index and apoptosis indes were employed to measure cell apoptosis.

Results

The results showed that Ganoderma spores reduced the splenic index to different degrees in each group and the best effect was seen in the high dose group (P<0.05). Terminal dexynucleotidyl transferase (TdT)-mediated 2′-Deoxyuridine 5′-Triphosphate nick end labeling staining revealed that the apoptotic index in all groups administered Ganoderma spores differed significantly from the model group, and a dose-response was observed. Flow cytometric analysis indicated that spleen lymphocyte apoptosis in the model group was extensive. Each dose of Ganoderma spores inhibited dexamethasone-induced apoptosis in spleen lymphocytes, and a dose-response was observed as well. The highest dose of Ganoderma spores decreased Malondialdehyde content in serum induced by dexamethasone (P<0.05).

Conclusion

The findings imply that the pretreatment of the mice with Ganoderma spores could reduce the apoptosis rate induced by dexamethasone in their splenic lymphocytes.     

Effects of Ganoderma applanatum polysaccharide on cellular and humoral immunity in normal and sarcoma 180 transplanted mice

Ganoderma applanatum polysaccharide (GAPS) was extracted and purified from Polyporaceae Ganoderma applanatum (Pers.) Pat. It was found that in vitro GAPS (65.2–250 μg/mL) increased the spleen cell proliferation response to ConA; GAPS (500 μg/mL) enhanced IL-2 production of spleen cells stimulated with ConA; GAPS (125–500 μg/mL) augmented the IFNγ production of spleen cells stimulated with ConA. Administering GAPS (20 mg/kg/day s.c.) to normal mice for 10 consecutive days enhanced spleen cell proliferation responses to ConA and the IL-2 and IFNγ production of spleen cells stimulated with 5 μg/mL ConA. It also increased spleen cell primary antibody responses to sheep red blood cells. Administering GAPS (20 mg/kg/day, s.c.) to sarcoma 180 transplanted mice for 15 consecutive days restored the NK activity and the IL-2 and IFNγ production of the spleen cells which were suppressed by the tumour.     

赤芝子实体化学成分的研究

从多孔菌科灵芝属植物赤芝子实体中分离得到1个化合物,经化学和光谱方法鉴定为麦角甾-7,22(E)-二烯-3-酮。     

Antihepatoma Activity of the Acid and Neutral Components from Ganoderma lucidum

Ganoderma lucidum extracts or isolated components have been shown previously to acquire many potential biochemical and pharmacological activities, including cancer preventive or antitumor effects. The supercritical fluid extracts of Ganoderma lucidum (total component, TC) and its acid component (AC) and neutral component (NC), were evaluated in vitro and in vivo for their antihepatoma activities. The NC showed a conspicuous inhibitory effect on tumor growth of Heps‐bearing mice, whereas AC was less effective. The TC, NC and AC all inhibited the proliferation of BEL‐7402 cells through apoptosis pathway and cell cycle arrest. Additionally, the NC and TC induced cell cycle arrest at the G2/M phase, but the AC resulted in a marked increase in the percentage of cells at G1 phase by flow cytometry. It is suggested that NC is an indispensable effective component in terms of antihepatoma activity and its constituents need to be investigated in detail. It was found that the NC, which was detected by GC‐MS, contained fatty acids and steroids; hence, it is proposed that some compounds such as long‐chain fatty acids and steroids in the NC might also contribute to the antihepatoma activity, although the anticancer activities of G. lucidum traditionally have been considered to be associated with triterpenoids. Copyright © 2012 John Wiley & Sons, Ltd.     

Immunomodulation of RAW264.7 macrophages by GLIS, a proteopolysaccharide from Ganoderma lucidum

The immunomodulatory effect of Ganoderma lucidum immunomodulating substance (GLIS) on macrophages has been investigated as part of on-going research into the anti-cancer properties of Ganoderma lucidum. Proliferation of bone marrow macrophages (BMMs) was enhanced by GLIS in a dose-dependent manner. Microscopic examination revealed that numerous GLIS-treated RAW264.7 macrophages were enlarged and formed pseudopodia. Exposure of RAW264.7 macrophages to GLIS resulted in significant increases in NO production, induction of cellular respiratory burst activity, and increased levels of IL-1beta, IL-12p35 and IL-12p40 gene expression. Our data indicate that GLIS activates the immune system by modulating cytokine production.     

灵芝螺旋藻片免疫调节功能研究

目的:观察灵芝螺旋藻片增强免疫力的功能作用。方法:经口给予0.083、0.83、2.5g/kg BW剂量的灵芝螺旋藻片30d,测定迟发型变态反应、抗体生成细胞数、半数溶血值和单核-巨噬细胞廓清影响试验。结果:灵芝螺旋藻片能提高小鼠的半数溶血值、小鼠抗体生成细胞数,即能增加体液免疫功能,能增强小鼠的迟发型变态反应,及增加细胞免疫功能;能提高小鼠单核-巨噬细胞的碳廓清能力,增强非特异性免疫功能。结论:御坊堂牌灵芝螺旋藻片具有免疫调节功能作用。     

飞船搭载灵芝的生理生化研究

为了探明宇宙飞船搭载灵芝的空间生物学效应,评价搭载灵芝发生的变异,本研究分别以搭载与地面对照灵芝原基和子实体为材料,用NBT光化还原法测定SOD酶活力,聚丙烯酰胺电泳分析了它们的酯酶同工酶和过氧化物同工酶。结果显示飞船搭载对四个灵芝菌株的影响不同:在SOD酶活力上,不管是原基还是子实体,菌株Sx比对照Cx都有较明显增加;其他几个菌株与对照相比也有变化。电泳结果显示,S4与C4、Sh与Ch两组的原基、子实体的酯酶同工酶差异不明显。而这两个时期的S3与C3、Sx与Cx的酯酶同工酶酶谱差异比较明显。灵芝菌株的过氧化物同工酶酶带数较少,除了在原基期Sx比Cx少一条酶带外,其余菌株与对照都没有差异。这些结果表明,经飞船搭载后,Sx、S3与地面对照相比变化较大,而S4、Sh与对照相比基本没有变化。     

飞船搭载灵芝的生理生化研究

为了探明宇宙飞船搭载灵芝的空间生物学效应，评价搭载灵芝发生的变异，本研究分别以搭载与地面对照灵芝原基和子实体为材料，用NBT光化还原法测定SOD酶活力，聚丙烯酰胺电泳分析了它们的酯酶同工酶和过氧化物同工酶。结果显示飞船搭载对四个灵芝菌株的影响不同：在SOD酶活力上，不管是原基还是子实体，菌株Sx比对照Cx都有较明显增加；其他几个菌株与对照相比也有变化。电泳结果显示，S4与C4、Sh与Ch两组的原基、子实体的酯酶同工酶差异不明显。而这两个时期的S3与C3、Sx与Cx的酯酶同工酶酶谱差异比较明显。灵芝菌株的过氧化物同工酶酶带数较少，除了在原基期Sx比Cx少一条酶带外，其余菌株与对照都没有差异。这些结果表明，经飞船搭载后，Sx、S3与地面对照相比变化较大，而S4、Sh与对照相比基本没有变化。     

Ganoderma lucidum mycelia enhance innate immunity by activating NF-kappaB

Ganoderma lucidum is a popular medicinal mushroom in China and Japan for its immunomodulatory and antitumor effects. The goal of this research is to investigate the effect of dried mycelia of Ganoderma lucidum produced by submerged cultivation on the enhancement of innate immune response. We found that Ganoderma lucidum mycelia (0.2-1.6 mg/ml) stimulated TNF-alpha and IL-6 production after 8h treatment in human whole blood. IFN-gamma release from human whole blood was also enhanced after 3 day-culture with Ganoderma lucidum mycelia (0.2-1.0mg/ml). However, Ganoderma lucidum mycelia did not potentiate nitric oxide production in RAW264.7 cells. To better understand the possible immuno-enhancement mechanisms involved, we focused on nuclear factor (NF)-kappaB activation. Electrophoretic mobility shift assay revealed that the Ganoderma lucidum mycelia (1.6 mg/ml) activated kappaB DNA binding activity in RAW264.7 cells. These results provide supporting evidences for the immunomodulatory effect of Ganoderma lucidum mycelia.     

不同栽培因素对灵芝化学成分含量的影响

对不同栽培条件下所产的国药灵芝进行了人体必需氨基酸、总氮量及无机元素含量的比较测定，结果表明：菌材可能是影响灵芝化学成分含量的主要因素，不同产地栽培的药材有差异，不同的采收期限也会影响其结果。     

赤芝孢子的三萜和甾体类成分研究

目的研究破壁赤芝(Ganoderma lucidum)孢子中的化学成分.方法用硅胶柱色谱法及Sephadex LH-20柱色谱法进行分离纯化,依据理化性质和光谱数据鉴定化合物结构.结果从破壁赤芝孢子的乙酸乙酯提取物中分离鉴定了8个化合物,分别被鉴定为灵芝酸A(Ⅰ)、甲基灵芝酸A(Ⅱ)、甲基灵芝酸B(Ⅲ)、灵芝酸C2(Ⅳ)、灵芝酸G(Ⅴ)、麦角甾-7,22-二烯-3β,5α,6β-三醇(Ⅵ)、麦角甾醇过氧化物(Ⅶ)和麦角甾-7,22-二烯-3β-十五酸酯(Ⅷ).结论化合物Ⅱ、Ⅲ、Ⅶ和Ⅷ为首次从赤芝孢子中分离得到.     

雪灵芝总皂苷对胃肠平滑肌的作用

目的：观察雪灵芝总皂苷是否具有促进小鼠肠蠕动的作用以及对家兔离体肠肌活动的影响。方法：1．肠蠕动实验：先给雪灵芝总皂苷、甲基硫酸新斯的明、蒸馏水30min后,再给予碳末。给予碳末30min后测定碳末在肠中的推进率。2．兔子离体肠肌实验：测离体兔肠肌给药前和给药后的活动情况,研究雪灵芝总皂苷对离体肠肌活动的影响。结果：雪灵芝总皂苷高、中剂量组有明显的增加肠蠕动的作用（P〈0．05）,对家兔离体肠肌给药后与给药前比较,张力和振幅有显著差别（P〈0．05）。结论：雪灵芝总皂苷能增加小鼠肠蠕动,对离体兔肠肌有明显的收缩作用。