不同产地丹参脂溶性成分指纹图谱的比较研究

目的：利用聚类分析对各地野生及家种丹参的指纹图谱数据进行分析。了解不同地理区域野生丹参以及同一地区家种丹参的遗传差异,为丹参药材优良品种的筛选提供依据。方法：采用高效液相色谱法,以甲醇-水系统为流动相进行梯度洗脱,流速1．0mL／min。检测波长254nm,使用SPSS11．0数据处理软件对所得指纹图谱中考察指标进行聚类分析。结果：采用无性繁殖的相同产地的家种丹参指纹图谱具有很好的相似性,相应的品系药材可以归入同一类;而野生丹参则不可按其地理区域归类。结论：采用无性繁殖丹参的遗传稳定性较好。     

泰山白首乌生物学特性及繁育技术

泰山白首乌为萝摩科植物耳叶牛皮消 (CynanchumauriculatumRoyleexWight)的块根 ,药用或生产淀粉食用。在海拔 70 0米以上泰山阴坡 ,少有分布 ,现处于野生状态 ,为开发泰山白首乌这一珍贵植物资源。本文主要介绍了该生物学特性和繁育技术。     

丹参MSAP分子标记技术体系的优化及其应用

以陕西省野生丹参为材料利用正交设计对MSAP预扩增和选择性扩增体系中关键因素优化筛选,以建立适合丹参的MSAP反应体系,并用于5个野生丹参居群表观遗传多样性分析.结果表明:25μL MSAP双酶切反应体系中加入EcoRⅠ和MspⅠ各10 U,37℃酶切8 h,酶切较充分;最佳预扩增反应体系25μL,包含连接产物2.5μL,Mg2+(25 mmoI/L)1.0μL,dNTPs(2.5 mmol/L)2.0μL,Taq酶(5 U/μL)0.4μL,上下游引物E00/HM00(50ng/μL)各1.0μL;最佳选择性扩增反应体系25μL,包含稀释100倍的预扩增产物1.5 μL,Mg2+(25 mmol/L)2.0μL,dNTPs(2.5 mmol/L)2.0μL,Taq酶(5 U/μL)0.1μL,上下游引物E-ACG/HM-CAA(50 ng/μL)各0.8μL.选用5对引物组合对5个野生丹参居群的50个单株进行表观遗传多样性分析,平均DNA甲基化多态性谱带为95.48%;UPGMA聚类分析结果表明,50个丹参单株在相似性系数0.58处可分为3大类.     

泰山虫草的研究进展

对泰山虫草菌Cordyceps taishanensis的生物学特性、泰山虫草原寄生体昆虫的生物学特性、泰山虫草发生区的生态环境、泰山虫草与冬虫夏草有效成分的比较、泰山虫草的药理作用、泰山虫草发酵培养基及发酵条件的研究和利用替代寄生体昆虫培养泰山虫草等七个方面对泰山虫草的研究进展进行了具体阐述.分析了目前存在的问题及今后的研究重点领域.     

泰山顶上的野生园林植物资源与利用

山地野生园林植物是发展园林绿化事业的物质基础和种质保存、品种栽培的珍贵资源 ,有着广阔的引种、开发前景。文章分析了泰山顶上的野生园林植物的特点、种类、资源、分布、筛选出 2 0种有重要研究、栽培和开发价值的多功能性树种 ,并就如何进行保护、引种、发展和综合利用提出建议和措施。     

泰山项上的野生园林植物资源与利用

山地野生园林植物是发展园林绿化事业的物质基础和各质保存、品种栽培的珍贵资源,有着广阔的引种、开发前景。文章分析了泰山项上的野生园林植物的特点、种类、资源、分布、筛选出２０种有重要研究、栽培和研发价值的多功能性树种,并就如何进行保护、引种、发展和综合利用提出建议和措施。     

泰山地区药用植物资源现状调查与分析

目的:调查泰山地区药用植物资源状况,为有效保护并合理利用当地药用植物资源提供科学依据。方法:结合全国第四次中药资源普查,通过野外调查、种植基地调查、市场调查及查阅相关文献资料,对该地区药用植物资源展开调查记录。结果:此次调查泰山地区植物类中药资源97科431种,涉及国家重点调查品种28种,水生、耐盐中药资源42种,通过与第三次中药资源普查有关资料核实,发现泰山地区新纪录物种6种。结论:调查结果显示泰山地区中药资源种类丰富,但野生药材的种类和数量均有所下降,需注意加强保护以利于资源的可持续性。     

泰山赤鳞鱼胚胎发育的研究

泰山赤鳞鱼学名多鳞白甲鱼(Scaphesthes macrolepisBleeker),隶属鲤形目、鲤科、鲃亚科、白甲鱼属,是多鳞白甲鱼分布在泰山的一个地理种群,适合生长于清澈的山涧溪流中。赤鳞鱼肌肉味美不腥,含有12种以上的矿物质和微量元素、18种以上的脂肪酸以及丰富的人体必需氨基酸,具有良好的抗衰老及预防多种疾病的作用[1—3],是山东省重点保护的唯一淡水鱼类。近些年来随着消费市场对名特优水产品种类和数量日益增长,泰山赤鳞鱼需求量逐年大幅度上升。但另一方面由于气候变化、泰山降雨减少、旅游资源开发、人工过度捕捞等因素的影响,泰山赤鳞鱼野生资源急剧减少,濒临灭绝。因此,开展泰山赤鳞鱼苗种人工繁育是一项十分紧迫的工作。对泰山赤鳞鱼胚胎发育时序和特征进行研究,可为泰山赤鳞鱼的保护和开发利用提供基础依据。1材料与方法试验鱼取自山东农业大学泰山赤鳞鱼研究中心,人工养殖条件下自然产卵受精。大约在五月中旬,水温上升到18—19℃时,雌鱼开始产卵,受精卵被细沙埋住。取出后,在水族箱中进行孵化。随时清除死卵,每日换水两次,换水量为1/2,以保证充足的氧气供应。本实验共对10批次不同亲鱼的受精卵进行观察,定期取样,取卵安置于培...     

丹参水溶性成分的地域分布特点分析

分析全国不同产地丹参中水溶性成分的含量变化情况及其地域分布特点。采用HPLC法测定不同产地丹参水溶性成分的含量,运用SAS统计软件分析丹参水溶性成分相关性及其地域分布特点。回归分析表明,丹参素与丹酚酸A含量呈负相关,原儿茶醛与迷迭香酸含量呈负相关,咖啡酸与丹参素含量呈负相关,丹酚酸B与丹酚酸A、丹参素含量呈正相关。聚类分析表明,三个野生药材与山东和河南大部分产地、四川中江栽培药材水溶性成分较高聚为一类。建议丹参药材中丹参素≥0.14%,原儿茶醛≥0.01%,咖啡酸≥0.01%,迷迭香酸≥0.17%,丹酚酸B≥3.0%,丹酚酸A≥0.01%。丹参的水溶性成分呈现地域性差异,传统道地产区的含量较高。     

干旱胁迫对丹参叶片气体交换和叶绿素荧光参数的影响

研究了干旱处理15d后,大叶型丹参和小叶型丹参2个品种幼苗气体交换和叶绿素荧光参数的变化.结果表明:在干旱胁迫15d后,大叶型丹参叶片净光合速率(Pn)和PSⅡ最大光化学效率(Fv/Fm)分别下降了66.42%和10.98%,而小叶型丹参的Pn和Fv/Fm分别下降了29.32%和5.47%,干旱胁迫对大叶型丹参Pn和Fv/Fm的影响明显大于小叶型丹参.小叶型丹参Pn下降主要由气孔因素造成,而大叶型则主要由非气孔因素所致.干旱胁迫使丹参叶片的气孔导度(Gs)下降,但明显诱导了水分利用效率(WUE)、非光化学猝灭系数(qN)和光呼吸速率与净光合速率比率(Pr/Pn)的增加,以提高干旱胁迫抗性.其中小叶型丹参的增幅明显大于大叶型丹参.表明小叶型丹参的抗干旱胁迫能力更强.     

腐殖酸对丹参生长的促进作用及其机理的研究

本文旨在研究腐殖酸对丹参生长的促进作用及其机理以提高丹以的产量和品质。水培试验研究表明：20ppm－80ppm腐殖酸促进了丹参根干物质的积累,80ppm时达到了最大,丹参根重比对照增加了49％,其作用机理是：通过提高根系活力,增加根系吸收能力,促进谷氨酸合成酶的活性、提高氮同化的效率,促进丹参根系分泌酸性磷酸酯酶以及改变体内酸性和中性磷酸酯酶的活性,以充分利用内的磷而适应低磷环境。     

Protective effects of Danshensu from the aqueous extract of Salvia miltiorrhiza (Danshen) against homocysteine-induced endothelial dysfunction

Homocysteine (Hcy) is a by-product of methionine metabolism. An imbalance of Hcy in the body may lead to hyperhomocysteinemia, a condition with elevated Hcy concentration in blood that may be one of the risk factors responsible for the development of several vascular diseases (thromboembolism, atherosclerosis, stroke, vascular diseases and dementia). Radix Salvia miltiorrhiza (Danshen), a well-known Chinese medicinal herb that can activate and improve blood microcirculation, is noticeable for its beneficial effect in treating cardiovascular diseases. The present study is to demonstrate the protective effect of Danshen extract against the homocysteine-induced adverse effect on human umbilical vein endothelial cell (HUVEC). Homocysteine (5 mM) not only decreased the cell viability but also caused the disruption of capillary-like structure formation in vitro. The protective effect of Danshen aqueous extract and its active compounds on endothelial cell function were demonstrated through an in vitro tube formation assay, which mimics the new blood vessel formation. To identify the active components in the aqueous extract of Danshen, the content was characterized by instrumental analysis using high performance liquid chromatography with diode array detector (DAD) and electrospray tandem mass spectrometry (ESI-MS/MS). Interestingly, Danshen extract and its pure compounds showed different effectiveness in protecting HUVEC against Hcy-induced injury according to the following descending order: Danshen aqueous extract, 3-(3,4-dihydroxy-phenyl)-2-hydroxy-propionic acid (Danshensu), protocatechuic acid, catechin and protocatechualdehyde. We believed that such findings might provide evidence in understanding the beneficial effects of Danshen on the cardiovascular system.     

Influence of ethanol concentration of extraction solvent on metabolite profiling for Salviae Miltiorrhizae Radix et Rhizoma extract by 1H NMR spectroscopy and multivariate data analysis

Salviae Miltiorrhizae Radix et Rhizoma (Danshen in China) and its related preparations are widely used in clinical practice due to its high medicinal value. In recent years, ¹H NMR technology has made great progress and demonstrated its unique advantages in the field of botanical metabolomics. In this study, ¹H NMR-based metabolomics was used to investigate the dissolution of various metabolites in Danshen as a function of ethanol concentration. ¹H NMR spectroscopy of Danshen extract identified 28 metabolites including 6 sugars, 11 amino acids, 3 organic acids, 4 salvianolic acids, and 4 tanshinones. Multivariate statistical analysis was used to classify and compare various Danshen extracts. PCA and HCA were used to obtain a global overview of the similarity in the samples and two-class OPLS-DA models were established for identifying characteristic metabolites. Then, ¹H-qNMR method was used to estimate the concentration of 22 metabolites, which is helpful to further describe the changes in metabolite ratios of various Danshen extracts. The result of this study laid the foundation for further biological activity research, and also provided an important reference for subsequent process research and quality control of Danshen related preparations.     

藏红花对肝纤维化大鼠肝组织TGF-β1 表达的影响

目的:通过观察藏红花对肝纤维化大鼠肝组织TGF-β1表达的影响,探讨藏红花预防肝纤维化作用的机制。方法:清洁级SD雄性大鼠60只随机分为正常组、模型组、藏红花组、丹参组,每组十五只。应用30%四氯化碳橄榄油溶液3ml/kg.腹腔注射制备肝纤维化大鼠模型。治疗8周后,通过HE和Masson染色观察肝纤维化的形成、免疫组化检测肝组织中TGF-β1蛋白的表达。结果:臧红花组、丹参组与模型组相比肝纤维化程度均轻于模型组(P<0.01),而藏红花组与丹参组相比肝纤维化程度无显著性差异(P>0.05),臧红花组、丹参组与模型组相比TGF-β1蛋白的表达均明显减少,而藏红花组与丹参组相比TGF-β1蛋白的表达没有显著性差异(P>0.05)。结论:藏红花能有效减轻肝纤维化大鼠的肝脏损伤及纤维化程度,其机制可能与抑制肝内TGF-β1的表达,抑制HSC的激活以及阻断HSC与TGF-β1之间的恶性循环有关。     

PXR-mediated transcriptional activation of CYP3A4 by cryptotanshinone and tanshinone IIA

Danshen (Radix Salvia miltiorrhiza) is a famous Traditional Chinese Medicine used widely for the treatment of coronary heart disease and cerebrovascular disease. Diterpenoid tanshinones including tanshinone I, tanshinone IIA and cryptotanshinone are the major bioactive components from Danshen herb. Previous reports have demonstrated that Danshen extracts could induce the expression of CYP3A in rodents, however, the constituents responsible for Danshen-mediated CYP3A induction and the underlying molecular mechanisms remain unknown. The discovery of a family of nuclear receptors such as pregnane X receptor (PXR), constitutive androstane receptor (CAR) and glucocorticoid receptor (GR) gives insight into the molecular explanation of CYP3A induction by xenobiotics. In the present study, interactions between Danshen constituents and human PXR were evaluated using a reporter gene assay. Our observations showed that Danshen ethanol extract could activate human PXR and induce the CYP3A4 reporter construct in HepG2 cells. Tanshinone IIA and cryptotanshinone were identified as efficacious PXR agonists, and cryptotanshinone activated the CYP3A4 promoter more strongly than tanshinone IIA. Furthermore, CAR and GR were also involved in the induction of CYP3A4 expression by tanshinones, though their roles seemed not as important as PXR. Treatment of LS174T cells with cryptotanshinone or tanshinone IIA resulted in a significant increase of CYP3A4 mRNA, which was consistent with the results from the reporter gene assay. Collectively, activation of PXR and the resultant CYP3A4 induction mediated by cryptotanshinone and tanshinone IIA provide a molecular mechanism for previously observed CYP3A induction by Danshen extracts, and our findings also suggest that caution should be taken when Danshen products are used in combination with therapeutic drugs metabolized by CYP3A4.     

复方丹参注射液在新生儿疾病中的应用

目的：介绍复方丹参注射液在新生儿疾病中的应用情况。方法：通过检索近年来公开发表的相关文献归纳而成。结果和结论：复方丹参注射液在新生儿疾病中具有较好的效果和应用前景。     

Investigation of cytochrome P450 1A2 and 3A inhibitory properties of Danshen tincture

Danshen (Salvia miltiorrhiza Bunge) as a famous Traditional Chinese medicine is widely used in the treatment of cardiovascular and cerebrovascular diseases in the world. Danshen tincture (DT), extracted from Danshen root with a mixture of water and alcohol, is a commonly used preparation method for human consumption. The aim of this study was to investigate the effects of DT on the cytochrome P450 (CYP) 1A2 and 3A activities by human and rat liver microsomes. Effects of DT were assessed with use of Danshen ethanolic extract (DEE) and selective substrates, markers of CYP activities. DEE (0.5-10 μg/ml) competitively inhibited human and rat liver microsomal CYP1A2 activity with inhibition constant (K(i)) values at 3.40 and 5.16 μg/ml, respectively. At the same time, DEE (2.5-20 μg/ml) not only noncompetitively inhibited human liver microsomal CYP3A4/5 activity with a K(i) of 11.9 μg/ml, but also competitively inhibited rat liver microsomal CYP3A1/2 activity with a K(i) of 52.1 μg/ml. The data indicate that DEE inhibited the metabolism of CYP1A2 and 3A substrates in human and rat liver in vitro with different mode of inhibition. This study may be helpful for clinical application of Danshen tincture.     

Danshensu is the major marker for the antioxidant and vasorelaxation effects of Danshen (Salvia miltiorrhiza) water-extracts produced by different heat water-extractions

Some of the major components of Danshen (Salvia miltiorrhiza), a widely used Chinese herbal medicine rich in phenolic acids, are thermosensitive and may degrade to other phenolic acids during extractions with heating. The chemical profiles of Danshen water-extract may vary with different heat water extraction at different temperatures, affecting the composition and bioactivity of the extracts. In this study, six water-extracts of Danshen obtained from heat reflux water extraction and microwave-assisted extraction with water (MAE-W) at different temperatures were tested for their composition and pharmacological effects. Among these extracts, the third-round MAE-W (100 °C) extract had the highest phenolic acids and tanshinones contents, with the strongest antioxidant activity in 2,2-diphenyl-1-(2,4,6-trinitrophenyl) hydrazyl (DPPH) assay and ferric reducing/antioxidant potential (FRAP) assay. This extract also showed the strongest inhibitory effects on 2,2′-azobis-2-amidinopropane (AAPH)-induced hemolysis in human red blood cells, hydrogen peroxide-induced apoptosis in rat heart H9c2 cells and the highest relaxation effects on rat basilar artery. The antioxidant effects of Danshen water-extracts linearly correlated to their relaxation effects (r = 0.895–0.977). Through multiple linear regression analysis, danshensu was found to be the most significant marker in the antioxidant and vasodilation effects of Danshen water-extract, while tanshinone IIA as the marker on hydrogen peroxide-induced apoptosis in rat heart H9c2 cells. Danshensu is, therefore, a useful marker for the quality control of Danshen water-extracts in antioxidant and vasodilation, while tanshinone IIA for anti-apoptotic potential of different extracts.     

丹参对冻融入胎儿卵巢组织异种移植早期血管生成的影响

目的：研究冻融人胎儿卵巢组织移植早期血管生成过程中微血管形态和密度的改变以及血管生成相关基因mRNA的表达;探讨丹参注射液对移植物血管生成的影响。方法：冻融胎儿卵巢组织异种移植至裸鼠肾被膜下,按给药不同分为对照组（生理盐水）和丹参组（丹参注射液每只0．09g／d）,分别于移植后48h、7d和28d回收移植物。结果：移植后两组卵巢组织微血管密度均明显增多;对照组移植后7d血管密度达峰,丹参组血管密度在移植后48h即已显著上升,此后两个时间段保持相对平稳。Angiopoietin-2mRNA表达在移植后48h两组均显著升高,丹参组上升幅度大于对照组（P〈0．05）。结论：冻融人胎儿卵巢组织新血管生成开始于异种移植后48h内,移植后7d组织内微血管密度达峰。丹参在移植早期应用可以促进移植后血管生成,其机制可能与它增加了血管生成相关因子Ang-2mRNA的表达有关。     

Magnesium as NMDA receptor blocker in the traditional Chinese medicine Danshen

Aqueous extracts of the traditional Chinese medicine Danshen, the dried roots of Salvia miltiorrhiza Bunge (Labiatae), blocked N-methyl-D-aspartate (NMDA) evoked currents in cerebrocortical neurons in vitro. The block of the NMDA-evoked currents was voltage dependent and showed the negative slope conductance reminiscent of the effect of Mg2+ ions. Atomic absorption spectrophotometry (AAS) revealed that aqueous Danshen extracts contained approximately 9mM magnesium. Fractionation of the extracts by high performance liquid chromatography followed by patch clamp recording and AAS indicated that magnesium ions were present in two distinct fractions. One fraction contained approximately 5 mM magnesium and blocked NMDA-induced currents indicating that it contained mostly free Mg2+ ions, while a second fraction did not possess NMDA antagonist activity despite the presence of approximately 4 mM magnesium suggesting that Mg2+ in this fraction was mostly chelated. Following removal of the free Mg2+ by ion exchange chromatography, the previously observed block of the NMDA-induced currents was abolished. These data demonstrate that Danshen contains both free and chelated Mg2+. Free Mg2+ ions account for the NMDA antagonist activity of Danshen in vitro.