对不同地区灵芝特性的激光拉曼光谱表征

采用激光拉曼光谱和荧光光谱分析方法对灵芝（Ganoderma lucidum（W．Curt．：Fr．）Karst．）的特性进行表征与研究,特别是对不同地区生长的灵芝菌肉及菌管的发育状况进行比较、并进行对应的光谱表征比较研究,给出一些有价值的结果。     

刺五加-灵芝双向固体发酵工艺优化及抗氧化活性评价

发酵中药是传统中药材精细高值化利用的有效途径,可以起到增效减毒、扩大适用范围等作用。通过双向固体发酵技术将药食同源的刺五加与灵芝有机结合,达到协同增效的目的。首先,通过单因素实验考察常规固体发酵参数的最适范围和微生物生长因子种类;其次,通过Box-Behnken设计和响应曲面优化,得到并验证最优的生长因子组合;最后,通过DPPH自由基清除能力、超氧化物歧化酶的活力和总抗氧化能力对发酵前后的刺五加提取物进行活性评价。结果表明,刺五加-灵芝双向发酵的最佳工艺为：粉碎粒度为10目,基质加水量为40%,菌丝体与固体基质的质量比为2∶1;最佳生长因子组合为：MnCl₂为84 mmol/kg、NaCl为56 mmol/kg、CaCl₂为149 mmol/kg;发酵后的刺五加提取物抗氧化活性显著增强,明显高于刺五加、灵芝及刺五加-灵芝混合提取物,提高了刺五加的保健功效,为系列大健康产品的开发提供了新的活性物质基础。     

高压热水提取灵芝多糖及对其抗氧化活性的影响

以灵芝子实体超微粉为原料,研究了提取温度、提取时间和料液比在高压热水条件下对灵芝多糖提取以及多糖提取液对DPPH自由基清除率的影响,并采用正交试验优化了灵芝多糖高压热水提取工艺。确定较优工艺条件为料液比1∶50g/mL、提取温度125℃下提取30min,一次高压热水提取灵芝多糖提取率达到4.54%。抗氧化实验结果表明,灵芝多糖对 DPPH 自由基有一定的清除能力,且与多糖质量浓度存在一定的量效关系。     

Comparison of the antitumor activity of polysaccharides extracted by boiling water and enzyme assistance from Ganoderma lucidum

Polysaccharides are the most important pharmacologically active constituents of Ganoderma lucidum. In this work, polysaccharides were extracted from Ganoderma lucidum with boiling water method and enzyme assisted method. The human liver hepatocellular carcinoma cell line HepG2 was used to compare the an- titumor effect of the two kinds of extraction with 3-（4, 5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） test. Both of these two kinds of Ganoderma lucidum polysaccharides reduced cell viability of cancer cell HepG2 in a dose and time-dependent manner. At low concentrations, there was no significant difference in the effectiveness of L 1 and L2; while at concentrations over 0.8 ug/mL, the difference in the effectiveness of L2 in comparison to L1 became significant. At the concentrations of 3.2 ug/mL, the cancer cells were almost killed in 2 d.     

荧光光谱法测定灵芝中多糖含量

以无水葡萄糖为标准品,对用荧光光谱法测定灵芝中多糖的含量进行了探讨,确定最佳实验条件为12.00mL浓硫酸,o.17mL 5%的苯酚溶液,室温下静置50min,荧光参数λex=478nm,λem=510nm.在此条件下对6种不同种灵芝的多糖含量进行了测定,多糖浓度与生成物的荧光强度呈良好的线性关系,线性方程为y=486...     

灵芝液体发酵产纤维素酶的提取及性质分析

通过对比实验,找出了较适合灵芝液体发酵并产生纤维素酶的PDY培养基：马铃薯200g,葡萄糖20g,酵母膏5g,加水至1000mL,并对纤维素酶的酶活力进行了测定。发酵液经过硫酸铵盐析,纤维素酶被纯化了1．61倍,通过快速液相色谱的分离纯化,酶被纯化了1．96倍,酶活分别达到5．68和6．89。经过快速液相色谱分离纯化,有6个组分峰。其中第一、第二峰分离较好,含量较高。对灵芝纤维素酶的氨基酸组成进行了分析,灵芝纤维素酶并经紫外-可见光吸收光谱和红外吸收光谱对纤维素酶进行了分析。     

灵芝抑制肿瘤微环境中Treg细胞功能的机制

为探究灵芝抑制肿瘤的可能机制,分别采用灵芝三萜、多糖及三萜多糖配伍对H22荷瘤小鼠模型进行干预,检测瘤体重量、体积,流式细胞法检测肿瘤组织中Treg细胞比例,RT-PCR法检测趋化因子CCL22、受体CCR4和FOXP3 mRNA表达水平,IHC法检测FOXP3蛋白表达水平,ELISA法检测IL-10、TGF-β1、IL-2含量.结果显示,灵芝能抑制H22肿瘤生长,配伍组抑瘤率(47.8%)明显高于三萜组(21.4%)和多糖组(30.8%);灵芝能降低肿瘤微环境中Treg细胞比例,配伍组比例(1.93%)明显低于三萜组(3.04%)和多糖组(2.18%);灵芝能显著降低相关mRNA的表达、FOXP3和IL-10、TGF-β1水平,增加IL-2水平.因此,灵芝可通过减少Treg细胞向肿瘤组织浸润、抑制其分泌细胞因子,同时增加IL-2水平来促进抗肿瘤免疫,且三萜和多糖合用可能存在协同作用.     

Study on Quality Control of Compound Anoectochilus roxburghii (Wall.) Lindl. by Liquid Chromatography–Tandem Mass Spectrometry

Compound Anoectochilus roxburghii (Wall.) Lindl. (A. roxburghii) oral liquid (CAROL) is a hospital preparation of A. roxburghii and Ganoderma lucidum (G. lucidum), which have hepatoprotective effects. Eight active components (five nucleosides/nucleobases and three triterpenoid acids) in CAROL, A. roxburghii, and G. lucidum were simultaneously detected by high-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS). The multiple reaction monitoring (MRM) mode was applied for the detection of analytes. These eight compounds were separated well within 12 min and quantified using the internal standard working curve method. The method showed good linearity (R² > 0.9935) and high sensitivity (limit of detection = 0.29 ng/mL). The analyte recovery ranged from 85.07% to 97.50% (relative standard deviation < 3.31%). The content of the target analytes in four batches of CAROL, and the raw materials of G. lucidum and A. roxburghii from the five regions was determined using this method. The contents of guanosine and ganoderic acid A in four batches of oral liquid were high and stabilized and could be recommended as quality markers (Q-marker) for CAROL. Simultaneous qualitative and quantitative analysis of nucleosides and triterpenoid acids in CAROL, A. roxburghii, and G. lucidum by LC–MS/MS based on the MRM model was reported for the first time. The proposed method provides a sensitive, rapid, and reliable approach for the quality control of Chinese medicinal products.     

弱碱性介质下微波法提取灵芝多糖的优化

为了探寻一种高效的灵芝多糖提取方法,采用单因子分析和正交实验,对弱碱性介质下微波法提取灵芝多糖进行了优化研究.各因素对灵芝多糖提取的影响依次为提取时间,pH值,固液质量比,预浸时间.最适条件为预浸1.5 h,提取15 min,固液质量比1∶110,pH 8.0,提取2次.灵芝多糖提取率为3.200%,结果表明:该方法能显著提高灵芝多糖的提取率.     

两种灵芝硒多糖分离纯化及性质鉴定

从灵芝（Ganoderma lucidum)加硒深层培养的菌丝（含Se 1600μg/g）中,经沸水、碱沸水提取,醇析,透析,Sevage法脱蛋白,DEAE－cellulose柱层析纯化后,得SeGLP-1和SeGLP-2两种灵芝硒多糖。经SephadexG-100、聚丙烯酰胺凝胶龟泳和紫外光谱分析鉴定,SeGLP-1和SeGLP-2为均一级分。GC与PC分析表明：SeGLP-1含5种单糖Gal、Man、Gle、Xyl、Rha,n(Gal):n(Xyl):n(Gle):n(Rha):n(Man)=0.23:0.72:1.00:0.25:0.09.SeGLP-2含有4种单糖Gal、Xyl、Glc、Rha,n(Gal):n(Xyl):n(Glc):n(Rha)=25.59:16.30:1.00:0.86。经红外光谱分析,确定SeGPL-1和SeGLP－2均是由α-糖苷键连接的吡喃多糖。