Isolation, purification, and partial characterization of prunellin, an anti-HIV component from aqueous extracts of Prunella vulgaris

Prunellin, an anti-HIV active compound, was isolated from aqueous extracts of the Chinese medicinal herb, Prunella vulgaris, and purified to chromatographic homogeneity. Infrared and NMR spectroscopy identified prunellin as a polysaccharide. Elemental analyses, precipitation with calcium(II), barium(II), or 9-aminoacridine suggest a sulfated polysaccharide. Paper chromatography of the exhaustively hydrolyzed material indicates the presence of glucose, galactose, xylose, gluconic acid, galactonic acid and galactosamine as the constituent monosaccharides. The molecular size of prunellin, as determined by gel permeation chromatography and the Squire method on Sephadex G-75, is about 10 kDa.     

仙鹤草多糖的分离纯化及理化性质研究

目的从中药仙鹤草中提取仙鹤草多糖,并对其理化性质进行研究。方法利用水提醇沉法得粗多糖,并依次经纤维素DE-52柱和Superdex-200凝胶柱色谱进行纯化。采用光谱法和色谱法对其总糖含量、单糖组成、相对分子质量等理化性质进行研究。结果分离纯化的仙鹤草多糖HAPⅢ总糖含量、蛋白含量、糖醛酸含量分别为81%,3.4%,45%,由半乳糖、葡萄糖、鼠李糖、阿拉伯糖、甘露糖、岩藻糖、木糖组成,摩尔比为5.65∶2.67∶2.61∶1.86∶1.12∶1.09∶1。其平均相对分子质量为1.90×105。结论仙鹤草多糖HAPⅢ是一水溶性酸性杂多糖。     

不同产地夏枯草指纹图谱研究

目的建立不同产地夏枯草药材的指纹图谱,为夏枯草药材的质量控制提供依据。方法色谱柱:Agilent 5HC-C_(18)(250 mm×4.6 mm,5μm);流动相:乙腈(A)-0.1%磷酸水溶液(B),梯度洗脱;流速:1.0 mL·min~(-1);检测波长:210 nm;柱温:30℃。结果建立了夏枯草药材的指纹图谱,标示了18个共有峰。16批药材中,有14批药材指纹图谱与对照图谱相似度达到0.88以上,表明大多数产地的夏枯草药材质量一致性较好。结论该方法稳定、可靠、重复性好,可以有效地用于夏枯草药材的质量控制。     

夏枯草中苯丙素和三萜的分离和鉴定

从夏枯草中用色谱法分离得到1个新的苯丙素类化合物1和5个已知的三萜类化合物2－6,经波谱法和化学法鉴定为3,4,α-三羟基苯丙素丁酯（1）,2α,3α,24－三羟基乌苏－12,20（30）－二烯－28－酸（2）、2α,3α,24－三羟基齐墩果－12－烯－28－酸（3）、2α,3α,24－羟基乌苏－12－烯－28－酸（4）、2α,3β－二羟基齐墩果－12－烯－28－酸（5）和2α,3β－二羟基乌苏－12－烯－28－酸（6）,5个三萜类化合物均为首次从夏枯草中分离得到。     

Isolation and characterization of an anti-complementary protein-bound polysaccharide from the stem barks of Eucommia ulmoides

EWDS-1, a homogeneous protein-bound polysaccharide, was isolated as an anti-complementary agent from the stem barks of Eucommia ulmoides. EWDS-1 was identified as a branched proteoglycan with average molecular weight about 2,000,000 Da, composed of Gal, Glc and Ara in the ratio of 2.1:1.0:0.9, along with trace of Rha, Xyl, Man, as well as 3.95% of protein. The linkages of the residues of EWDS-1 were deduced by methylation analysis and NMR technique. Bioassay showed that EWDS-1 inhibited complement activation on both the classic and alternative pathways with CH(50) and AP(50) values of 203+/-20 microg/ml and 45+/-8 microg/ml, respectively. Preliminary mechanism studies by using complement component depleted-sera indicated that EWDS-1 inhibits activation of complement system by interacting with C1q, C1r, C1s, C2, C3, C4, C5 and C9. The results suggested that EWDS-1 could be of promising benefits in treatment of the complement associated diseases.     

Anti-allergic and anti-inflammatory triterpenes from the herb of Prunella vulgaris

The activity-guided fractionation of the extract of the herb of Prunella vulgaris (Labiatae) led to the isolation of four triterpenes, i.e., betulinic acid, ursolic acid, 2 alpha,3 alpha-dihydroxyurs-12-en-28-oic acid, and 2 alpha-hydroxyursolic acid. One of these compounds, 2 alpha,3 alpha-dihydroxyursolic acid, demonstrated significant inhibition on the release of beta-hexosaminidase from the cultured RBL-2H3 cells in a dose-dependent manner; the IC50 value was calculated to be 57 microM. When the isolated compounds were tested for their effects on the production of nitric oxide from cultured murine macrophages, RAW 264.7 cells, ursolic acid and 2 alpha-hydroxyursolic acid exhibited strong inhibitory activities (IC50 values, 17 and 27 microM, respectively).     

Isolation and characterization of an anti-complementary polysaccharide D3-S1 from the roots of Bupleurum smithii

The preliminary data from hemolytic assays indicated that the hot-water extract of the roots of Bupleurum smithii had anti-complementary activity. Further bioactivity-guided fractionation led to the isolation of D3-S1, a homogeneous form of acidic polysaccharide. D3-S1 was a branched polysaccharide with average molecular weight about 2,000,000 Da, composed of Ara, Gal and GalA in the ratio of 2.6:1.0:1.2, along with trace of Rha, Glc, Xyl and Man. Methylation analysis and NMR identified the linkages of the residues of D3-S1. Functional analysis showed that D3-S1 inhibited complement activation on both the classic and alternative pathways with CH(50) value of 0.34+/-0.02 mg/ml and AP(50) value of 0.081+/-0.003 mg/ml, respectively. Preliminary mechanism studies by using complement component depleted-sera indicated that D3-S1 selectively interacts with C1s, C3 and C4, but not C1q, C1r, C2, C5 and C9. The results suggested that D3-S1 could be of potential benefits in treatment of the complement-associated diseases.     

夏枯草中水溶性多糖的分离纯化及化学研究

目的研究夏枯草中水溶性多糖组分.方法采用DEAD琼脂糖凝胶柱色谱进行纯化,化学和光谱方法进行结构特征分析.结果从夏枯草沸水提取物中分离得到5个多糖部位,其中XKC00、XKC02-A、XKC02-B均为均一分子量多糖,分子量分别为2.6x 10(4),1.36x 10(5),5 600 Da.结论XKC00为首次从夏枯草中分离得到的一个中性杂多糖.     

夏枯草中齐墩果酸和熊果酸的含量测定方法研究

目的通过采用高效液相色谱法,测定不同产地夏枯草中齐墩果酸和熊果酸的含量,比较夏枯草药材质量的优劣。方法色谱柱:Kromasil-C_(18)(250 mm×4.6 mm,5μm);流动相:乙腈-甲醇-0.2%磷酸水(5∶82∶13)等度洗脱;流速:1.0 mL·min~(-1);检测波长:210 nm;柱温:25℃。结果齐墩果酸在0.041~0.410 mg·mL~(-1)与峰面积呈现良好的线性关系(r=0.9999),平均回收率为99.1%,RSD为1.8%;熊果酸在0.054~0.540 mg·mL~(-1)与峰面积呈现良好的线性关系(r=0.9998),平均回收率为98.1%,RSD为2.4%。结论该方法简单、准确、可靠,为夏枯草的质量控制与评价提供了有效的手段。     

白蔹多糖PAJM-I的分离纯化和结构分析

目的 从白蔹中提取分离白蔹多糖,并研究其结构性质.方法 经热水提取、乙醇沉淀、Sevag法脱蛋白后得到粗多糖PAJM,进一步用DEAE-Sapharose Fast Flow, Sephadex G-200分离纯化得到PAJM-I,对分离到的主要多糖应用高碘酸氧化、高效凝胶渗透色谱(HPGPC),GC,GC-MS,IR等方法进行结构分析.结果 提取纯化的白蔹多糖(PAJM-I)是均一的多糖,相对分子量为3.25×104,鼠李糖,木糖,甘露糖和丰乳糖组成摩尔比为:12.98:23.52:45.06:18.44.分子中1??3,1??2,1??6连接的糖苷键数比为51%:26%:23%.结论 首次从白蔹中分离得到PAJM-I均一多糖,且PAJM-I含有α-D-葡萄吡喃糖.     

HPLC测定夏枯草中的芦丁和槲皮素

目的 采用HPLC法测定夏枯草中芦丁和槲皮素的含量.方法 色谱柱为Alltima C_(18)(250MM×4.6 mm,5 μm),流动相为甲醇-0.1%冰醋酸,检测波长350 nm,流速1.0 mL·min~(-1),柱温30℃.结果 芦丁、槲皮素的线性范围分别为0.0149～0.2380 mg·mL~(-1)(r=0.9999)、0.0019～0.0306 mg·mL~(-1)(r=0.9998),平均回收率分别为97.78%(RSD=0.83%)、101.18%(RSD=0.83%).安徽、湖北、贵州、江西、广西五产地中芦丁含量分别为0.3527、0、0.2337、2.1164、0.9578 mg·g~(-1)生药,槲皮素含量分别为0.0872、0.0473、0.0721、0.5219、0.1755 mg·g~(-1)生药.结论 所建方法操作简便、准确,可作为夏枯草中芦丁和槲皮素的含量测定方法.     

夏枯草化学成分研究

采用柱色谱分离技术,从夏枯草全草70%乙醇浸膏的乙酸乙酯和正丁醇部位中分离得到14个化合物,经MS和NMR分析鉴定为:白桦脂酸(1)、乌苏酸(2)、2α,3α-二羟基-12-烯-28-乌苏酸(3)、2α,3β-二羟基-12-烯-28-乌苏酸(4)、2α,3β,24-三羟基-12-烯-28-乌苏酸(5)、2α,3α,19-三羟基-12-烯-28-乌苏酸(6)、齐墩果酸(7)、槲皮素-3-O-β-D-葡萄糖苷(8)、槲皮素-3-O-β-D-半乳糖苷(9)、山奈酚-3-O-β-D-葡萄糖苷(10)、芦丁(11)、丹参素甲酯(12)、丹参素乙酯(13)、迷迭香酸乙酯(14).其中化合物5、6、10、13为首次从该植物中分离得到,化合物5、13为首次从该属植物中分离得到.     

Isolation and characterization of an immunomodulatory protein from bovine colostrum

A colostral protein that augments natural killer (NK) cell activity was isolated from bovine colostrum. This protein, designated matemal immunity enhancing factor (MIEF), increased NK cell-mediated cytotoxicity against human tumor targets when added to cultures of resting peripheral blood lymphocytes. The NK cell stimulatory activity of the MIEF was demonstrated at the concentrations as low as 0.1–0.01 μg/ml. Purified MIEF showed an apparent molecular weight of 22,000 in SDS-polyacrylamide gel electrophoresis. The unusual biochemical characteristics of the MIEF distinguish it from other cytokines. The MIEF was soluble at a cold temperature, and precipitated by raising the temperature. This thermal precipitability was reversible, and dependent on the concentration, pH, and ionic strength. Maximal precipitation was observed at neutral pH, and higher ionic strength.     

灵芝多糖分离鉴定及抗肿瘤活性的研究

目的探讨灵芝多糖的分离纯化、理化性质及初步抗肿瘤活性。方法采用热水提取 ,乙醇分级沉淀 ,DEAE 32离子交换柱色谱等方法从灵芝子实体中分离得到灵芝多糖。用SephadexG 10 0凝胶柱色谱法测定其纯度和平均分子量。用薄层色谱及气相色谱法测定其单糖组成。溴化四唑蓝法测定其初步体外抗肿瘤活性。结果从灵芝子实体中分离得到 4种均一多糖 (GLPL1 ～GLPL4 ) ,其中GLPL1 和GLPL3为主要成分 ,其分子量为 4 10 0和 5 70 0。GLPL1 由葡萄糖组成 ;GLPL3由葡萄糖和半乳糖组成。GLPL1 和GLPL3对人鼻咽癌细胞增殖有显著的抑制作用 ,GLPL3还对人胃癌细胞、人结肠癌细胞增殖有一定的抑制作用。结论灵芝多糖GLPL1 、GLPL3是很有潜力的抗肿瘤或辅助抗肿瘤药物     

Changes in bioactive components related to the harvest time from the spicas of Prunella vulgaris

Context: Prunella vulgaris L. (Labiatae) is a perennial plant common in China and Europe and is rich in rosmarinic acid (RA), ursolic acid (UA), and oleanolic acid (OA). The dried spica of P. vulgaris has been used as traditional medicine in China for over a hundred years. To our best knowledge, no study has been conducted to determine the influence of harvesting time on concentrations of bioactive compounds of P. vulgaris. Objective: In the current study, changes in the bioactive compounds present in spicas were investigated at five harvest times over 2 months. Materials and methods: Plant material were collected at five fixed dates: 5th May, 20th May, 7th June, 15th June, and 25th June and assayed for chemical contents by high-performance liquid chromatography (HPLC). Results: Among the different harvest times, the highest levels of RA (56.81 mg·g(-1)), UA (2.77 mg·g(-1)), and OA (0.91 mg·g(-1)) were found on 5th May, whereas the lowest levels of RA (1.66 mg·g(-1)), UA (2.27 mg·g(-1)), and OA (0.43 mg·g(-1)) were observed on 25th June. Discussion and conclusion: As each medicinal product has its own content requirement for different bioactive components, the optimum harvest time might be determined according to the accumulation dynamics of target compound in dried spicas of P. vulgaris. These results may be useful for determining the optimal harvest time when bioactive components are at the maximum level, which is in early May.     

Changes in bioactive components related to the harvest time from the spicas of Prunella vulgaris

Context: Prunella vulgaris L. (Labiatae) is a perennial plant common in China and Europe and is rich in rosmarinic acid (RA), ursolic acid (UA), and oleanolic acid (OA). The dried spica of P. vulgaris has been used as traditional medicine in China for over a hundred years. To our best knowledge, no study has been conducted to determine the influence of harvesting time on concentrations of bioactive compounds of P. vulgaris.

Objective: In the current study, changes in the bioactive compounds present in spicas were investigated at five harvest times over 2 months.

Materials and methods: Plant material were collected at five fixed dates: 5th May, 20th May, 7th June, 15th June, and 25th June and assayed for chemical contents by high-performance liquid chromatography (HPLC).

Results: Among the different harvest times, the highest levels of RA (56.81 mg·g^?1), UA (2.77 mg·g^?1), and OA (0.91 mg·g^?1) were found on 5th May, whereas the lowest levels of RA (1.66 mg·g^?1), UA (2.27 mg·g^?1), and OA (0.43 mg·g^?1) were observed on 25th June.

Discussion and conclusion: As each medicinal product has its own content requirement for different bioactive components, the optimum harvest time might be determined according to the accumulation dynamics of target compound in dried spicas of P. vulgaris. These results may be useful for determining the optimal harvest time when bioactive components are at the maximum level, which is in early May.     

夏枯草抗肿瘤活性成分及其作用机制研究进展

夏枯草作为一味临床常用中药,具有清肝泻火、明目、散结消肿的功效,对甲状腺癌、乳腺癌、肝癌等多种癌症表现出较强的抗肿瘤作用。夏枯草提取物及其有效成分可通过多种途径发挥抗肿瘤作用,其主要途径包括诱导细胞凋亡、抑制细胞侵袭及转移、抑制细胞增殖、诱导自噬、抗肿瘤血管生成、逆转肿瘤多药耐药性和调节免疫功能等。并通过调控miRNA和Wnt/β-catenin、PI3/Akt、AMPK/mTOR/ ULK1、PI3K/Akt/mTOR、RANKL/ RANK/ OPG、PI3K/PTEN/NIS/AKT/mTOR等信号通路,从分子机制上验证了夏枯草提取物具有抗肿瘤作用。本文对夏枯草提取物抗肿瘤的作用机制进行综述,以期为其进一步的研究和应用提供参考。     

RP-HPLC法同时测定夏枯草中3种三萜酸的含量

目的:建立同时测定夏枯草中齐墩果酸、熊果酸和2α,3α-二羟基-12-烯-28-熊果酸含量的方法。方法:采用反相高效液相色谱法。色谱柱为Diamonsil C18(2)柱(250mm×4.6mm,5μm),流动相为甲醇-1%醋酸铵水溶液(85∶15,V/V),流速为1.0mL·min-1,检测波长为210nm,柱温为30℃。结果:齐墩果酸、熊果酸和2α,3α-二羟基-12-烯-28-熊果酸的进样量分别在0.1～2.0μg(r=0.9998)、0.3～6.0μg(r=0.9995)和0.04～0.80μg(r=0.9997)范围内与各自峰面积积分值呈良好的线性关系;三者平均加样回收率分别为99.0%、98.6%和98.7%,RSD分别为0.9%、1.4%和1.1%(n均为9)。结论:该方法准确、简便、重复性好,可用于夏枯草的质量控制。