蒙古黄芪组织培养及同源四倍体的诱导与鉴定

通过蒙古黄芪组织培养技术的正交试验，确定了蒙古黄芪最佳繁殖培养基：MS＋BA0．6mg／L＋IAA0．2mg／L＋PP3330．2mg／L；叶作为诱导愈伤组织的外植体优于茎，最适培养基：MS＋BA2．0mg／L＋KT1．0mg／L＋NAA0．5mg／L；最佳生根培养基：1／2MS＋IAA0．5mg／L＋IBA2．0mg／L＋ABT2．0mg／L。建立了蒙古黄芪快速繁殖体系；在无菌条件下利用秋水仙碱诱导蒙古黄芪，获得62个染色体加倍遗传稳定的变异试管苗株系，通过试管苗根尖染色体鉴定，对诱导获得的蒙古黄芪染色体加倍株系进行初步筛选，为进一步获得蒙古黄芪优良品系奠定基础。     

蒙古黄芪的研究进展

目的介绍最近几年来蒙古黄芪的研究概况及展望其进一步研究价值。方法以近几年发表的国内外文献为依据,从化学成分和药理作用等方面对蒙古黄芪的研究进展做一综述。结果蒙古黄芪含有多种化学成分,药理活性显著,具有良好的深入研究和开发的价值。     

蒙古黄芪化学成分研究

目的 研究豆科黄芪属植物蒙古黄芪的化学成分及结构.方法 用硅胶柱和C18反相柱,重结晶等方法分离,并通过理化性质和核磁共振等方法确定化合物的结构.结果 从蒙古黄芪根中分到7个化合物,分别鉴定为,Nepehinone(Ⅰ)、黄芪皂苷Ⅱ(Ⅱ)、黄芪皂苷甲(Ⅲ)、异黄芪皂苷(Ⅳ)、(6aR,11aR)-9,10-二甲氧基紫檀烷葡萄糖苷(Ⅴ)、2'-羟基-3',4'-二甲氧基-异黄烷葡萄糖苷(Ⅵ)、芒柄花素(Ⅶ).结论 化合Ⅰ为首次从黄芪属植物中分离得到.     

蒙古黄芪化学成分研究

目的研究蒙古黄芪的化学成分。方法采用硅胶柱色谱、凝胶柱色谱方法进行分离纯化,利用波谱分析法结合理化性质确定化合物结构。结果从蒙古黄芪的乙酸乙酯和正丁醇萃取部分中共分离得到14个化合物,分别鉴定为(6aR,11R)-9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷(1),(3R)-2′-羟基-3′,4′-二甲氧基异黄烷-7-O-β-D-葡萄糖苷(2),5′-羟基-3′-甲氧基异黄酮-7-O-β-D-葡萄糖苷(3),芒柄花苷(4),(6aR,11R)-3-羟基-9,10-二甲氧基紫檀烷(5),(3R)-7,2′-二羟基-3′,4′-二甲氧基异黄烷(6),5′,7-二羟基-3′-甲氧基异黄酮(7),芒柄花素(8),黄芪甲苷(9),黄芪皂苷II(10),蔗糖(11),腺嘌呤核苷(12),十六烷酸单甘油酯(13),十六烷酸(14)。结论化合物5,7,11～14为首次从黄芪属植物中分离得到,化合物6为首次从蒙古黄芪中分离得到。     

蒙古黄芪化学成分研究

目的研究蒙古黄芪的化学成分。方法采用硅胶柱色谱、凝胶柱色谱方法进行分离纯化,利用波谱分析法结合理化性质确定化合物结构。结果从蒙古黄芪的乙酸乙酯和正丁醇萃取部分中共分离得到14个化合物,分别鉴定为（6aR,11R）-9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷（1）,（3R）-2′-羟基-3′,4′-二甲氧基异黄烷-7-O-β-D-葡萄糖苷（2）,5′-羟基-3′-甲氧基异黄酮-7-O-β-D-葡萄糖苷（3）,芒柄花苷（4）,（6aR,11R）-3-羟基-9,10-二甲氧基紫檀烷（5）,（3R）-7,2′-二羟基-3′,4′-二甲氧基异黄烷（6）,5′,7-二羟基-3′-甲氧基异黄酮（7）,芒柄花素（8）,黄芪甲苷（9）,黄芪皂苷II（10）,蔗糖（11）,腺嘌呤核苷（12）,十六烷酸单甘油酯（13）,十六烷酸（14）。结论化合物5,7,11～14为首次从黄芪属植物中分离得到,化合物6为首次从蒙古黄芪中分离得到。     

阿克苏黄芪与蒙古黄芪中皂苷成分的比较

目的　对新疆产阿克苏黄芪 (ArstragalusaksuensisBge)的总皂苷进行定性定量分析。方法　以山西产的蒙古黄芪 [Astragalusmembranaceus(Fisch)Bge .var.Mongholicus(Bge)Hsiao]为对照 ,比较阿克苏黄芪与蒙古黄芪皂苷类成分的异同。结果　阿克苏黄芪总皂苷含量为 4 713mg·g-1,蒙古黄芪总皂苷含量为 4 75mg·g-1。结论　二者总皂苷含量无显著差异。但所含皂苷成分不同 ,可用作二者的鉴别     

蒙古黄芪中黄芪甲苷的提取工艺

目的研究蒙古黄芪中黄芪甲苷的最佳提取工艺。方法运用正交实验设计方法，考察了乙醇浓度、加乙醇量、煎煮时间及煎煮次数等4个因素对有效成分黄芪甲苷提取率的影响，以黄芪甲苷含量作为检测指标，采用高效液相色谱法 蒸发光散射检测（HPLC-ELSD）法测定，并确定最佳提取工艺。结果提取黄芪中黄芪甲苷的最佳工艺条件为8倍量80%乙醇，回流提取2次，每次3 h。结论该提取工艺合理，质量稳定，重现性好，黄芪甲苷提取率高。     

蒙古黄芪化学成分的分离鉴定

从蒙古黄芪(Astragalus membranaceus Bge.var.mongholicus(Bge.)Hsiao)乙醇提取物中分离得到10个单体,分别鉴定为棕榈酸(Ⅰ),羽扇醇(Ⅱ),β-谷留醇(Ⅲ),黄芪皂甙Ⅳ(astragaloside Ⅳ,Ⅳ),黄芪异黄烷甙(3S-(—)mucronulatol-7-D-glucopyranoside,Ⅴ),胡萝卜甙(Ⅳ),α-联苯双酯(dimethyl 4,4-dimethoxy-5,6,5′,6′-dimethylenedioxybiphenyl-2,2-dicarboxylate,(Ⅶ),天冬酰胺(Ⅷ),γ-氨基丁酸(Ⅸ),蔗糖(Ⅹ)。其中Ⅰ,Ⅱ和Ⅸ首次由蒙古黄芪巾分得,联苯双酯(Ⅶ)为已知合成物,但在天然药物黄芪中属首次分出,黄芪异黄烷甙(Ⅴ)未见报道。     

蒙古黄芪化学成分的分离与鉴定

目的研究中药蒙古黄芪的化学成分。方法用硅胶柱、Sephadex LH 20和HPLC柱色谱方法进行分离纯化,通过理化性质和核磁共振氢谱、碳谱数据确定化合物的结构。结果从蒙古黄芪中分离得到5个化合物,分别鉴定为5,7,4′三羟基异黄酮(1)、4,2′,4′,羟基查尔酮(2)(、3R)-8,2′二羟基7,4′二甲氧基异黄烷(3)、(6αR,11αR)9,10二甲氧基紫檀烷3OβD-葡萄糖苷(4)、2′羟基3′,4′二甲氧基异黄烷7OβD葡萄糖苷(5)。结论其中化合物1、2为首次从黄芪属中分离得到。     

蒙古黄芪凝集素对K562细胞的增殖抑制和诱导凋亡

目的研究蒙古黄芪凝集素(AMML)对慢性髓系白血病细胞系K562的生长抑制作用及对细胞周期和凋亡的影响。方法用MTT法测定AMML对K562细胞的生长抑制作用;荧光染色和流式细胞术分析检测AMML诱导K562细胞周期变化和凋亡的影响。结果AMML对K562细胞的生长增殖具有明显的抑制作用,呈剂量和时间依赖性。60mg.L-1的AMML处理K562细胞72h后其生长抑制率高达89%。AMML处理的K562细胞呈现典型的细胞凋亡形态改变,如胞核破裂、染色质固缩。流式细胞仪(FCM)分析结果说明AMML可以引起K562细胞S期阻滞,并诱导K562细胞凋亡。结论AMML通过S期阻滞抑制K562细胞的增殖并诱导其凋亡,AMML在抗肿瘤新药开发中具有潜在的应用价值。     

蒙古黄芪的化学成分研究

目的 研究蒙古黄芪乙醇提取物石油醚、二氯甲烷、醋酸乙酯萃取部位的化学成分.方法 利用硅胶柱色谱、凝胶柱色谱、制备液相色谱等方法进行分离、纯化,根据化合物的理化性质和波谱数据鉴定其结构.结果 分离得到14个化合物,分别鉴定为红车轴草素(1)、芒柄花素(2)、芒柄花素-7-O-β-D-葡萄糖苷(3)、毛蕊异黄酮(4)、毛蕊异黄酮-7-O-β-D-葡萄糖苷(5)、(6aR,11 aR)-3-羟基-9,10-二甲氧基紫檀烷(6)、(6aR,11aR)9,10-二甲氧基紫檀烷-3-O-β-D-葡萄糖苷(7)、黄芪皂苷I(8)、乙酰黄芪皂苷I(9)、黄芪皂苷Ⅱ (10)、异黄芪皂苷Ⅱ (11)、腺嘌呤(12)、β-谷甾醇(13)、胡萝卜苷(14).结论 此次从蒙古黄芪中主要分离得到黄酮类和皂苷类化合物.     

Micellar electrokinetic chromatography for the quantitative analysis of flavonoids in the radix of Astragalus membranaceus var. mongholicus

The separation and determination of eight flavonoids in the radix of Astragalus membranaceus (Fisch.) Bge var. mongholicus (Bge.) Hsiao was achieved by using micellar electrokinetic chromatography (MEKC) with a diode array detector (DAD). The influence of background electrolyte (BGE) concentrations and pH, surfactant concentrations, organic solvents, temperature, and voltage on the separate procedure was systematically investigated. The optimal resolution was obtained with a micellar phase consisting of 100 mM sodium cholate, 25 % (v/v) acetonitrile, 20 mM Na2B4O7, and 20 mM H3BO3 buffer at pH 9.2 by using a fused silica capillary at + 25 kV and 25 degrees C. A high reproducibility and good linearity ( R2 > 0.9978) were obtained over the concentration range tested. The relative standard deviations (R.S.D.s) from intra-day and inter-day precision for injection were less than 4.42 %. Six plant samples from different locations were then analyzed for the flavonoids by this newly developed MEKC method.     

Structural features of a polysaccharide from Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao

A polysaccharide Astragalus polysaccharide (APS) was obtained from the boiling water extract of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao. The results of gas chromatography (GC) indicated that APS consisted of l-rhamnose (l-Rha), d-xylose (d-Xyl), d-glucose (d-Glc), and d-galactose (d-Gal) in the molar ratio of 1:4:5:1.5. Its molecular weight was determined to be 3.01 × 10⁵ by high-performance gel filtration chromatography. The results of ¹H NMR and ¹³C NMR spectral analysis indicated that APS had a linear backbone mainly consisting of 1,3-linked β-d-Gal residues with insertion of β-Glc, 1,6-linked α-Gal, 1,5-linked β-Xyl, 1,4-linked β-Gal, β-d-Gal, 1,2-linked α-Rha, 1,2,4-linked α-Rha residues. HSQC spectrum indicated that C-2 and C-6 may link with H.     

基于MaxEnt和ArcGIS的定西市蒙古黄芪适宜性区划研究

目的:对定西市蒙古黄芪进行生态适宜性区划研究。方法:以定西市1 001批蒙古黄芪为考察样本(以每个有蒙古黄芪栽培的自然村为采集单位进行样本采集),从定西市经济作物技术推广站中获取其经、纬度信息,并从《中药资源空间信息网格数据库》中获取其55项环境生态因子(包括气候、地形、土壤等)信息。结合经、纬度信息和环境生态因子信息,以75%样本为训练集建立最大熵模型(MaxEnt模型),筛选出主要生态因子,并以另外25%样本为验证集进行模型验证。随后,应用地理信息系统(ArcGIS)对蒙古黄芪的适宜生长区进行划分。结果:建立的MaxEnt模型预测效果良好(训练集和验证集样本受试者工作特征曲线下面积分别为0.970、0.968)。共筛选出海拔、降水量、温度等8个主要生态因子(贡献率总和为98.90%);结合分析系统发现,海拔在1 800～2 650 m、4月降水量在25～50 mm、最冷月最低温在-16～-8℃、最湿月降水量在95～110 mm、温度季节性变化标准差在70～80、12月平均气温在-6～-3 v、10月降水量在30～50 mm、12月降水量在0～10 mm为定西市蒙古黄芪适宜生长的环境...     

柱前衍生化–高效液相色谱法测定蒙古黄芪多糖中单糖的组成

目的测定蒙古黄芪多糖中的单糖组成。方法蒙古黄芪多糖样品用三氟乙酸溶液水解成单糖,用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化,采用HPLC法于245nm波长处检测各单糖。结果蒙古黄芪多糖由甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、阿拉伯糖组成,物质的量比分别为0.02∶0.05∶0.17∶1∶0.18。结论此方法简单、快速、重复性好,可用于蒙古黄芪多糖中单糖组成分析。     

A combination of Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, enhanced nitric oxide production in obstructed rat kidney

BACKGROUND: The persistent renal hemodynamic maladjustment caused by imbalances between vasoactivators predisposes the kidney to tubulointerstitial injury and ultimate interstitial fibrosis. The decoction (A&A) of a combination of roots of two Chinese herbs, Astragalus membranaceus var. mongholicus and Angelica sinensis, has shown antifibrotic effects in rats with chronic kidney diseases and improvement of renal blood flow in rats with acute ischemic renal injury. In the present study, we investigated the effects and possible mechanisms of A&A on vasoactivators in the process of renal interstitial fibrosis. METHODS: Male Wistar rats were randomly divided into sham, unilateral ureteral obstruction (UUO) and UAA (UUO plus A&A administration) groups. After oral administration of A&A (14 g/kg/d) for 3, 7 and 10 days, morphological changes were evaluated by HE, Masson and Sirius red staining technique. The levels of Ang-II, ET-1, and the activities of different nitric oxide synthases (NOSs) in renal homogenate were measured by radioimmunoassay. The nitrite concentration as nitric oxide (NO) production was measured using the Griess reagent. Western blot analysis and immunohistochemical staining were performed to determine the expressions of eNOS, nNOS, and iNOS in the kidney. The ability of scavenging reactive oxygen species (ROS) was evaluated by spectrophotometry. RESULTS: Morphological analysis showed severe interstitial mononuclear cells infiltration, tubular atrophy, renal fibrosis and collagen expression in kidneys of UUO group, which reduced by A&A administration (p<0.05, UAA vs. UUO group). The levels of Ang-II and ET-I were increased in obstructed kidneys, but not significantly changed after A&A administration. NO production did not change in obstructed kidney at day 3 but increased in day 7 and day 10. Administering A&A progressively increased NO production by 2.2, 1.2, and 1.2 fold at days 3, 7 and 10, respectively. The activities of constitutive NOS and iNOS were comparable between UUO group and sham group. In contrast, the activity of constitutive NOS was much higher in UAA than that of UUO rats, which increased 78%, 68% and 78% at days 3, 7 and 10 respectively, although the protein expression of eNOS, nNOS and iNOS in renal tissue had no change in UAA rats. The activities of scavenging ROS in UUO group were not significantly different from the sham group at days 3 and 7, but increased at day 10 (24.1+/-15.0 vs. 10.1+/-0.8 U/min/mg protein, p<0.05). After A&A administration, the activities of scavenging ROS were significantly increased at days 3 and 7 (51.5+/-17.9 vs. 11.7+/-7.4 U/min/mg protein, p<0.05; and 16.1+/-5.6 vs. 7.7+/-1.4 U/min/mg protein, p<0.05) respectively, comparing with the UUO group. CONCLUSION: The anti-fibrosis effects of A&A might be associated with enhancing NO production via eNOS activation and scavenging ROS, and in turn might improve ischemic microvasculature and attenuate interstitial fibrosis.