南、北五味子的化学成分和鉴别方法研究进展

五味子为木兰科Magnoliacene五味子亚科Schisandraceae五味子属和南五味子属植物的统称，五味子亚科只有五味子属和南五味子属二属，约60种植物，分布于东亚、东南亚及北美的南部。我国两属均产，约30种，其中五昧子属有20种，南五味子属有10种，两属均是我国最为重要的药用植物资源类群之一^[1-3]。     

基于电子舌技术的南、北五味子及其产地的鉴别研究

目的:采用电子舌技术对不同产地南、北五味子的味道进行定量表征,建立识别南、北五味子及其产地的方法,构建区分南、北五味子的判别模型。方法:收集不同产地的南、北五味子样品39批,运用电子舌技术对样品的味道进行检测,通过主成分分析(PCA)、判别因子分析(DFA)对获取的味道特征数据进行处理和分析。结果:采用PCA和DFA两种化学计量学方法,电子舌技术均能够有效区分北五味子和南五味子,DFA能有效区分南五味子和北五味子的不同产地,验证指数分别为93、98,而PCA仅能区分南五味子的不同产地,对北五味子的产地区分不适用。结论:电子舌可用于南、北五味子及其产地的鉴别,为中药的鉴别提供了新的思路与方法。     

Compositions and the in vitro antimicrobial activities of the essential oils of Achillea setacea and Achillea teretifolia (Compositae)

GC-MS analysis of the isolated essential oils from air-dried aerial parts of Achillea setacea and Achillea teretifolia, an endemic taxon, resulted in the identification of 51 constituents (79.8% of the total oil) and 42 constituents (87.1% of the total oil), respectively. Eucalyptol (1,8-cineole) was the major constituent of both oils studied (18.5 and 19.9%, respectively). The antimicrobial activities of the essential oils were individually evaluated against 14 microorganisms. Both oils exhibited inhibitory effects on Clostridium perfringens, Acinetobacter lwoffii and Candida albicans with a range of minimum inhibitory concentration values extended from 0.28 to 2.25 mg/ml. Camphor and their derivatives, borneol, terpinen-4-ol and eucalyptol (1,8-cineol) can be considered as the main antimicrobial constituents of the oils studied.     

从抗氧化角度评价南五味子"醋制入肝"的炮制机制

目的:从体内抗氧化和体外抗氧化相结合角度评价南五味子"醋制入肝"的炮制机制。方法:以抗坏血酸为阳性对照,采用紫外分光光度法,于517 nm处测定南五味子醋制前后样品对2,2-二苯基-1-苦味基肼(DPPH)吸光度的影响,考察南五味子醋制前后体外清除自由基能力差异;以四氯化碳(CCl4)致大鼠急性肝损伤为动物模型,SD大鼠随机分成正常对照组、CCl4模型组、五味子炮制前后给药组分别设置高、中、低3个剂量组,给药剂量以生药量计算分别为5,2.5,1.25 g.kg-1。连续给药10 d,每日1次。于造模后24 h剖取肝脏,测定南五味子醋制前后饮片高、中、低剂量组肝组织中总抗氧化能力(T-AOC)含量变化情况;体内外相结合综合评价南五味子醋制前后抗氧化能力差异。结果:体外实验结果显示,醋制南五味子清除DPPH自由基(IC501.887 1 g.L-1)的能力比南五味子原药材清除能力(IC504.571 2 g.L-1)强;体内总抗氧化能力测定结果显示五味子炮制前和炮制后各剂量组均可显着升高大鼠肝脏中T-AOC活性(与对照组比P<0.05),炮制后中剂量和小剂量组效果要明显优于炮制前相应剂量组(P<0.05)。结论:从抗氧化角度揭示醋制南五味子"醋制入肝"的炮制机制。     

华中五味子性别相关的分子标记建立

目的:建立华中五味子性别相关的RAPD标记。方法:以华中五味子雌雄株嫩叶为材料,用改良CTAB法分别提取基因组DNA,通过单因子与正交实验优化RAPD反应体系,对其性别的基因组差异进行研究。结果:25μL总体积:Mg2+浓度2.5 mmol/L,dNTPs浓度0.08 mmol/L,引物浓度0.6μmol/L,Taq酶1.5 U,DNA模板60 ng,退火温度41.3℃,循环次数35个。在400条随机引物中,只有S353筛选得到一条541 bp的雄性特异条带。结论:该标记可作为其性别鉴定依据。     

华中五味子野生抚育中的关键技术

目的:为保证南五味子药材基原物种华中五味子种群正常更新,野生南五味子药材资源的可持续利用及实现药材生产与生态环境保护协调发展提供参考。方法:采用野外居群观察、产区药农和经销商访谈、文献整理、设定基地用于观察和实践等方法,提出野生抚育关键技术。结果:关键技术包括通风透光、立架、人工补植、药材可持续采收及科学产地加工等五个方面。结论:本研究为指导产地南五味子药材的可持续采收、华中五味子野生资源的合理利用及药材质量的保障提供了技术支撑。     

Antigenotoxic and antioxidant activities of palmarosa and citronella essential oils

Ethnopharmacological relevance

Essential oils of palmarosa and citronella have been extensively used in ancient Indian and South-east Asian traditional medicines.

Aim of the study

These essential oils have been reported to exhibit antimicrobial, anti parasitic effects against bacteria, yeasts, filamentous fungi, and viruses. In the present study the oils were tested for their potential antigenotoxic and antioxidant properties in human lymphocyte cells.

Materials and methods

The antigenotoxic effect on human lymphocyte cells (measurement of cell viability, DNA damage) was studied using trypan blue dye exclusion test, plasmid pBR322 DNA strand scission, and comet assay. Anti-oxidant activity was evaluated by DPPH⁺ free radical scavenging and lipid peroxidation assay.

Results

The essential oils showed a good antigenotoxic activity against methyl methanesulphonate (MMS) and hydrogen peroxide. In addition, a significant dose dependent antioxidant activity was observed.

Conclusion

Our data provide evidence that support the usage of palmarosa and citronella essential oils in traditional herbal preparations. They can constitute a natural source of a new and safe antioxidant.     

华中五脂素--华中五味子种子中得到的新木脂素

目的：对华中五味子种子的化学成分进行研究，方法：运用各种色谱技术对其化学成分进行分离，波谱方法鉴定化学结构。结果：从华中五味子种子95％乙醇提取物的乙醚萃取物中分离得到华中五脂素(1)，安五脂素(2)，五味子酯甲(3)，五味子甲素(4)，顺酯酰戈米辛O(5)，6-O-苯甲酰戈米辛O(6)和五味子酮(7)7个木脂素类化合物．结论：化合物1为新的二芳基丁烷类木脂素，命名为华中五脂素；化合物6为首次从该植物中分离得到。     

Chemical compositions and anti-influenza activities of essential oils from Mosla dianthera

Ethnopharmacological relevance

Mosla dianthera as an aromatic herb is used in folk medicine for the treatment of cough, colds, fever, bronchitis, nasal congestion and headache.

Aim of the study

To characterize chemical compositions and to evaluate the anti-influenza effects of essential oils of M. dianthera (MDEO) in influenza virus A (IVA) infected mice.

Materials and methods

MDEO was obtained by hydrodistillation and analysed by gas chromatography-mass spectrometry (GC-MS). ICR mice were treated with MDEO for 5 consecutive days at doses of 90-360 mg/kg after post-infected. Levels of Serum IL-4 and IFN-γ were assayed by ELISA. Levels of MOD, SOD, TAOC and GSH-Px in lung tissue were determined by colorimetric method.

Results

GC-MS analysis revealed the presence of 29 components that account for 97.74% of phenolic sesquiterpenes and aromatic compounds. The major compounds were elemicin (16.51%), thymol (14.77%), β-caryophyllene (14.49%), iso-elemicin (9.22%), asarone (6.09%) and α-caryophyllene (5.26%). It had significant effects on decreasing lung viral titers, inhibiting pneumonia, reducing levels of serum IFN-γ and IL-4, and enhancing antioxidant activity in the lung tissue of IVA infected mice.

Conclusions

MPE could exhibit therapeutical effects in IVA infected mice as a suppressor of IVA replication and inflammatory mediators and a promoter of antioxidant potentials. Therefore, MDEO could provide a safe and effective therapeutic candidate for treatment of influenza and its subsequent viral pneumonia.     

Comparing chloroplast genomes of traditional Chinese herbs Schisandra sphenanthera and S. chinensis

ObjectiveSchisandra sphenanthera and S. chinensis are the two important medicinal plants that have long been used under the names of “Nan-Wuweizi” and “Wuweizi”, respectively. The misuse of “Nan-Wuweizi” and “Wuweizi” in herbal medical products calls for an accurate method to distinguish these herbs. Chloroplast (cp) genomes have been widely used in species delimitation and phylogeny due to their uniparental inheritance and lower substitution rates than that of the nuclear genomes. To develop more efficient DNA markers for distinguishing S. sphenanthera, S. chinensis, and the related species, we sequenced the cp genome of S. sphenanthera and compared it to that of S. chinensis.MethodsThe cp genome of S. sphenanthera was sequenced at the Illumina HiSeq platform, and the reference-guided mapping of contigs was obtained with a de novo assembly procedure. Then, comparative analyses of the cp genomes of S. sphenanthera and S. chinensis were carried out.ResultsThe cp genome of S. sphenanthera was 146 853 bp in length and consisted of a large single copy (LSC) region of 95 627 bp, a small single copy (SSC) region of 18 292 bp, and a pair of inverted repeats (IR) of 16 467 bp. GC content was 39.6%. A total of 126 functional genes were predicted, of which 113 genes were unique, including 79 protein-coding genes, 30 transfer RNA (tRNA) genes, and four ribosomal RNA (rRNA) genes. Five tRNA, four protein-coding genes, and all rRNA were duplicated in the IR regions. There were 18 intron-containing genes, including six tRNA genes and 12 protein-coding genes. In addition, 45 SSRs were detected. The whole cp genome of S. sphenanthera was 123 bp longer than that of S. chinensis. A total of 474 SNPs and 97 InDels were identified. Five genetic regions with high levels of variation (Pi > 0.015), trnS-trnG, ccsA-ndhD, psbI-trnS, trnT-psbD and ndhF-rpl32 were revealed.ConclusionWe reported the cp genome of S. sphenanthera and revealed the SNPs and InDels between the cp genomes of S. sphenanthera and S. chinensis. This study shed light on the species identification and further phylogenetic study within the genus of Schisandra.     

指纹图谱结合响应面法研究醋制南五味子炮制工艺

目的 优化南五味子醋制的炮制工艺.方法 以南五味子指纹图谱共有色谱峰峰面积的综合评分为指标,采用3因素3水平的Box-Behnken响应面实验设计,考察加醋量、闷润时间、蒸制时间不同因素对醋蒸南五味子炮制工艺的影响,优选最佳炮制工艺.结果 采用Design Expert 8.05软件对数据进行分析,建立综合评分与影响因子的多元二次回归方程.多元二次回归方程预测南五味子醋制最佳炮制条件为每100 g药材加醋量为28.10 g,闷润时间1.95h,蒸制时间3.90 h.结论 采用优选的炮制工艺能从整体化学成分角度保证醋制南五味子炮制质量.为制定其饮片的质量标准和临床应用提供依据.     

Chemical and biological evaluation of the essential oils of different Melaleuca species

The essential oils of the fresh leaves of M. ericifolia, M. leucadendron, M. armillaris and M. styphelioides were isolated by a hydrodistillation method and analysed by a gas chromatography/mass spectrometry (GC/MS) technique. The essential oil of M. ericifolia contained methyl eugenol (96.84%) as a major constituent, whereas M. leucadendron was rich in 1,8-cineole (64.30%). The essential oil of M. armillaris was rich in 1,8-cineole (33.93%) followed by terpinen-4-ol (18.79%), whereas M. styphelioides was rich in caryophyllene oxide (43.78%) and (-) spathulenol (9.65%). The essential oils of these species possessed antimicrobial and antifungal activities. M. ericifolia exhibited the highest inhibitory effects against Bacillus subtiles and Aspergillus niger. The antiviral activities of the essential oils against Herpes simplex virus type 1 (HSV-1) were studied in African green monkey kidney cells (Vero) by a plaque reduction assay. The volatile oil of M. armillaris was more effective as a virucidal (up to 99%) than that of M. leucadendron (92%) and M. ericifolia (91.5%). The effects of the essential oils on the antioxidant system status in carbon tetrachloride treated animals were studied. The essential oil of M. armillaris exhibited a marked antioxidant effect, it improved vitamin E, vitamin C and superoxide dismutase parameters so it can be used as a free radical suppressor.     

The chemical composition of some Lauraceae essential oils and their antifungal activities

The antifungal activity of Aniba rosaeodora, Laurus nobilis, Sassafras albidum and Cinnamomum zeylanicum essential oils were investigated against 17 micromycetes. Among the tested fungal species were food poisoning, spoilage fungi, plant and animal pathogens. In order to determine fungistatic and fungicidal concentrations (MIC and MFC) macrodilution and microdilution tests were used. Linalool was the main component in the essential oil of A. rosaeodora, while 1.8-cineole was dominant in L. nobilis. In sassafras essential oil safrole was the major component and in the oil of C. zeylanicum the main component was trans-cinnamaldehyde. The essential oil of cinnamon showed the strongest antifungal activity.     

南五味子总木脂素的催眠作用及对脑单胺类神经递质的影响

目的:对南五味子总木脂素(lignans from Schisandra sphenanthera Rehd.et Wils.,LSS)的催眠作用进行评价,并研究其对脑单胺类神经递质的影响。方法:南五味子总木脂素按250mg/kg、125mg/kg和62.5mg/kg剂量灌胃给予受试小鼠,每日1次,连续3日,末次给药后进行阈剂量和阈下剂量戊巴比妥钠睡眠试验,分别以睡眠潜伏期、睡眠时间和入睡动物数为指标;按上述剂量灌胃给予小鼠,每日1次,连续5日,末次给药后取脑,检测脑组织中去甲肾上腺素(norepinephrine,NE),5羟色胺(5-hydroxytryptamine,5-HT)和多巴胺(Dopamine,DA)浓度。结果:南五味子总木脂素各剂量均可延长阈剂量戊巴比妥钠小鼠睡眠时间,250mg/kg、125mg/kg剂量可增加阈下剂量戊巴比妥钠小鼠入睡动物数,250mg/kg剂量则可显著提高小鼠脑组织内5-HT浓度。结论:南五味子总木脂素有明显的促睡眠作用,其对脑组织中5-HT水平的提高可能与此有关。     

Butyryl- and acetyl-cholinesterase inhibitory activities in essential oils of Salvia species and their constituents

Extracts of Salvia (sage) species have been reported to have cholinergic activities relevant to the treatment of Alzheimer's disease. A lack of information on the inhibition of the enzyme butyrylcholinesterase, also considered to be a target in the treatment of the disease, prompted this in vitro investigation of the essential oils of S. fruticosa, S. lavandulaefolia, S. of ficinalis and S. of ficinalis var. purpurea for anti-butyrylcholinesterase activity. Dose-dependent inhibition of human cholinesterases by the extracts and constituents was determined using the method of Ellman. A time dependent increase in the inhibition of butyrylcholinesterase by the oils of S. fruticosa and S. of ficinalis var. purpurea was evident. IC(50) values decreased from 0.15 +/- 0.007 and 0.14 +/- 0.007 mg/mL after 5 min to 0.035 +/- 0.016 and 0.06 +/- 0.018 mg/mL after 90 min incubation time respectively. The slow onset of inhibition of butyrylcholinesterase was also shown by individual constituents, such as 3-carene and beta-pinene. Analyses of the chemical composition of the oils and anti-butyrylcholinesterase activity of their constituents revealed that none of the compounds tested would account for the total activity of the oils and that synergy is likely.     

Dibenzocyclooctadiene lignans from Kadsura polysperma and their antineurodegenerative activities

Eleven new dibenzocyclooctadiene lignans, polysperlignans A-K (1-11), and eight known analogues (12-19) were isolated from the stems of Kadsura polysperma. Their structures and absolute configurations were established using a combination of MS, NMR, CD, and single-crystal X-ray diffraction techniques. Selected compounds were evaluated for activity against β-amyloid- or hydrogen peroxide-induced neurotoxicity on PC12 cells, and 1, 2, 4, 5, 13, and 16 showed statistically significant neuroprotective effects in these in vitro assays.     

Effect of Wuzhi tablet (Schisandra sphenanthera extract) on the pharmacokinetics of paclitaxel in rats

Wuzhi tablet (WZ, registration no. in China: Z20025766) is a preparation of an ethanol herb extract of Wuweizi (Schisandra sphenanthera) containing 7.5 mg Schisantherin A per tablet. It was reported recently that WZ could significantly increase the blood concentrations of tacrolimus, which might be due to the inhibitory effect of WZ and its ingredients on P‐gp and/or CYP450 activity. Paclitaxel is a substrate of the efflux transporter P‐gp, and is mainly metabolized by CYP450 enzymes in the liver. Therefore, the purpose of this study was to investigate whether and how WZ affects the pharmacokinetics of paclitaxel in rats. After pretreatment with WZ, there were significant increases in the AUC_0‐24h of oral paclitaxel (from 280.8 ± 97.3 to 543.5 ± 115.2 h ng/mL; p < 0.05) and C_max (from 44.6 ± 16.4 to 86.8 ± 16.1 ng/mL; p < 0.05). The pharmacokinetic data for i.v. paclitaxel with WZ showed a relatively small (when compared against oral paclitaxel) but still significant increase in AUC_0‐24h (from 163.6 ± 22.1 to 212.7 ± 17.7 h ng/mL; p < 0.05) and a decrease in clearance (from 3.2 ± 0.6 to 2.2 ± 0.3 L/h/kg; p < 0.05). Thus, the presence of WZ improved the systemic exposure of paclitaxel in rats. The herb–drug interaction between WZ and paclitaxel should be taken into consideration in clinical use. Copyright © 2011 John Wiley & Sons, Ltd.     

6种代表性沉香精油的化学成分及抗氧化、抗炎活性比较分析

目的 比较不同来源、不同提取方式的6种沉香精油化合物组成、抗氧化能力及抗炎活性的差异。方法 采用气相色谱-飞行时间质谱联用仪（gas chromatography-time of flight mass spectrometry,GC-TOF MS）对6种沉香精油的化学成分进行分析,并测定其清除1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-trinitrophenylhydrazine,DPPH）与2,2-氮杂双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐[2,2-azabis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt,ABTS]自由基的能力,考察其对脂多糖（lipopolysaccharide,LPS）诱导的小鼠单核巨噬细胞RAW264.7活力的影响。结果 从通体香水蒸气蒸馏精油（TTS）、通体香超临界萃取精油（TTC）、板头香水蒸气蒸馏精油（BTS）、板头香超临界萃取精油（BTC）、奇楠沉香水蒸气蒸馏精油（QNS）和奇楠沉香超临界萃取精油（QNC）中鉴定出的化合物数量分别为48、55、59、71、72和35种,所含香味物质小分子芳香物质及倍半萜类成分相对含量之和分别为81.42%、87.84%、89.03%、62.05%、63.83%和28.36%,所含2-(2-苯乙基)色酮类化合物相对含量分别为0、0、0.25%、22.42%、17.48%、60.42%。6种沉香精油均具有清除DPPH和ABTS自由基能力,且质量浓度在0.5～5 mg/mL与自由基清除能力均呈正相关。3种水蒸气蒸馏沉香精油（BTS、TTS、QNS）对LPS诱导RAW264.7细胞的半数抑制浓度（half inhibitory concentration,IC₅₀）分别为18.61、30.30、18.05 μg/mL,而3种超临界CO₂萃取沉香精油（BTC、TTC、QNC）的IC₅₀值分别为41.98、38.45、24.50 μg/mL。结论 超临界萃取沉香精油同水蒸气蒸馏沉香精油化合物组成相差较大,其能够提取出更多的脂肪酸类和色酮类化合物;板头香和通体香采用同一提取方法所得精油化学成分相似,而奇楠沉香中含有更多含量的香味物质和2-(2-苯乙基)色酮类化合物。3种超临界萃取沉香精油清除DPPH自由基能力均强于水蒸气蒸馏沉香精油,而抗炎活性则相反,且3种沉香来源中,奇楠沉香精油抗炎能力最佳。     

星点设计-效应面法优选南五味子的提取工艺

目的:采用星点设计-效应面法优化南五味子的提取工艺。方法:以溶剂浓度,溶剂倍数及提取时间为自变量,五味子甲素提取率为因变量对自变量各水平进行多元线性回归和二项式拟合,用效应面法选择最佳提取工艺,进行预测分析,并将预测值与实验值进行比较。结果:二项式拟合复相关系数较高,r为0.9565,最佳提取工艺为62%的乙醇溶液回流提取2次,体积倍数为11.20倍,提取时间为3.00h。提取率预测值与实验值偏差为0.22%。结论:试验值与预测值吻合度高,星点设计-效应面法优化南五味子的提取工艺不仅方法简便,且预测性良好。     

Antibacterial and antioxidant activities in Sideritis italica (Miller) Greuter et Burdet essential oils

Sideritis italica (Miller) Greuter et Burdet is a widespread Lamiacea in the Mediterranean region used in traditional medicine. Essential oils were antibacterial against nine ATCC and as many clinically isolated Gram-positive and Gram-negative bacterial strains. Antibacterial activity was also found against Helicobacter pylori: a dose-dependant inhibition was shown between 5 and 25 microg/ml. The antibacterial activity of the oils was expressed as MICs (minimum inhibitory concentrations) and MBCs (minimum bactericidal concentrations). At a concentration between 3.9 and 250 microg/ml the oils showed a significant antibacterial effect against both Gram-negative and Gram-positive bacteria. In particular the ATCC strains Pseudomonas aeruginosa (MIC=3.9 microg/ml and 7.8 for flowerheads and leaves, respectively), Proteus mirabilis (MIC=15.6 and 7.8 microg/ml), Salmonella typhi (MIC=7.8 microg/ml) and Proteus vulgaris (MIC=15.6 microg/ml) were the most inhibited. Only Pseudomonas aeruginosa showed MBC at a concentration between 62.6 and 125 microg/ml. The antioxidant activity of the essential oils was evaluated by two cell free colorimetric methods: ABTS and DMPD; leaf oil is more active (4.29 +/- 0.02 trolox equivalents and 4.53 +/- 0.67 ascorbic acid equivalents by ABTS and DMPD, respectively). Finally the antioxidant activity of the essential oils was also evaluated by their effects on human whole blood leukocytes (WB) and on isolated polymorphonucleate (PMN) chemiluminescence. Comparing the effects of the oils from leaves and flowerheads on both PMN and WB chemiluminescence emission, we found no significant differences. Essential oils showed a dose-dependent and linear inhibitory activity on isolated PMN as well as on WB CL emission when PMA-stimulated. On the contrary, the inhibitory activity on resting cells was nonlinear. Our data represent an answer to the continual demand for new antibiotics and antioxidants for the continuous emergence of antibiotic-resistant strains and the growing interest in the substitution of synthetic antioxidants with natural ones. Furthermore, our finding of antibacterial and antioxidant activities in the essential oils from Sideritis italica flowerheads and leaves validated the use of the plant for medical purposes.