胚乳和外源赤霉素对五味子[Schisandra chinensis (Turcz.) Baill]种子发芽的影响

以组织培养和土壤播种的未经层积处理的五味子种子为材料，检测胚乳和外源赤霉素（GA3）对发芽影响的结果表明，在不加生长调节物质的MS培养基上，去掉外种皮带全胚乳的种子发芽率只有2．7％，而带1／2和1／3胚乳的种子发芽率则分别为46％和70．6％；培养基中添加GA，，可提高去掉外种皮带全胚乳和带1／2胚乳种子的发芽率，而对带1／3胚乳的种子则无效果。土壤播种时，用10—1000mg·L—GA3对去掉外种皮种子进行1~10d前处理，其中100mg·L—GA3前处理10d的发芽率最高，达到38．7％。

Effects of Different Decolorants on Retention Rate of Total Triterpenes in Fruit and Rattan Stems of Schisandra chinensis(Turcz.) Baill

This study was to investigate the decoloration effects of activated clay, activated carbon, diatomite, Ca group bentonite, kaolin, activated aluminum oxide, magnesium oxide and attapulgite. Ultraviolet-visible spectroscopywas adopted to de- tect the absorbanees of the extracts of Schisandra chinensis fruit and rattan stems, the contents of total triterpenoids in S. chinensis fruit and rattan stems were deter- mined, and the decoloration rate and the retention rate of total triterpenoids of S. chi- nensis fruit and rattan stems were calculated. The results indicated that, attapulgite exhibited the best decoloration effect on the extract of S. chinensis fruit, with a de- coloration rate up to 60.47%, activated carbon exhibited a better decoloration effect on the extract of S. chinensis rattan stems, with a decoloration rate up to 69.24%, and they had relatively higher retention rates of total triterpenoids. In the eight de- colorants, attapulgite showed the best decoloration effect on the extract of S. chi- nensis fruit, and activated carbon exhibited the best decoloration effect on the ex- tract of S. chinensisrattan stems.

Nortriterpenoids from the fruit stalk of Schisandra chinensis (Turcz.) Baill.

Objective:The fruit stalk of Schisandra chinensis(Turcz.)Baill.(S.chinensis)has been found to contain bioactive components similar to the fruit of S.chinensis.Here,we report a recent discovery about new nortriterpenoids with a novel skeleton and anti-gastric cancer activity,which were isolated from the fruit stalk of S.chinensis.Methods:The chemical components of ethyl acetate extract from 70%ethanol extract from S.chinensis fruit stalk were separated,purified,and identified by liquid chromatography methods(silica gel,ODS,HPLC)and extensive spectroscopic analyses(NMR,IR,UV,MS,CD).Results:Two new nortriterpenoids,schilancitrilactone M and 25-hydroxyl schindilactone D(1 and 2),along with ten known nortriterpenoids(3-12)were isolated from the fruit stalk of S.chinensis.The isolated compounds were tested for their cytotoxic activities against MGC-803 cells,and the results showed that compounds 6-8 possessed significant activities with IC50 of 9.01,11.77,and 2.74μmol/L,respectively.Conclusion:Twelve nortriterpenoids including two new compounds were isolated from the fruit stalk of S.chinensis for the first time.Among them,compounds 6-8 showed significant anti-gastric cancer activities.We postulated that the fruit stalk of S.chinensis could be used as an anti-gastric cancer drug.

Schisandra chinensis (Turcz.) Baill

Schisandra chinensis (Turcz.) Baill. is a deciduous woody vine, and belongs to the Schisandraceae family. This species is a horticultural plant with edible fruit. It is used medicinally and for making rope. It is distributed in Hebei, Heilongjiang, Jilin, Liaoning, Nei Mongol, Shanxi

Transcriptome-wide analysis of PIP reductase gene family identified a phenylpropene synthase crucial for the biosynthesis of dibenzocyclooctadiene lignans in Schisandra chinensis

Phenylpropenes,such as isoeugenol and eugenol,are produced as defend compounds,floral attractants,and flavor constituents by phenylpropene synthases belonging to the PIP reductase family.Moreover,isoeugenol is proposed to be involved in the biosynthesis of dibenzocyclooctadiene lignans,the main active compounds of Schisandra chinensis(Turcz.)Baill.fruits(SCF).S.chinensis,a woody vine plant,is widely used for its medicinal,horticultural,edible,and economic values.In this study,nine ScPIP genes were identified and characterized from the transcriptome datasets of SCF.The expression profiles revealed that ScPIP genes were differentially expressed during different developmental stages of SCF.Three ScPIPs were selected and cloned as candidate genes encoding phenylpropene synthases according to phylogenetic analysis.ScPIP1 was proved to function as isoeugenol synthase(IGS)and designated as ScIGS1 through in vivo functional characterization in Escherichia coli.Subcellular localization analysis demonstrated that ScIGS1 was localized in both the cytoplasm and nucleus.The three-dimensional(3D)model of ScIGS1 was obtained using homology modeling.Site-directed mutagenesis experiments revealed that the substitution of residues at positions 110 and 113 impacted the product specificity of ScIGS1 and the mutation of Lys157 to Ala abolishing catalytic function.Moreover,the kcat values of mutants were lower than that of ScIGS1 using a deep learning approach.In conclusion,this study provides a basis for further research on PIP reductases and the biosynthetic pathway of dibenzocyclooctadiene lignans.

指纹图谱技术结合化学计量学评价五味子醇提物质量探索研究

目的:优化五味子乙醇提取工艺,建立其醇提物HPLC指纹图谱并结合化学计量学综合分析其质量。方法:基于单因素试验,采用L9(34)正交试验设计,系统研究了乙醇浓度、提取时间、提取次数及料液比等因素对五味子中五味子醇甲和五味子乙素的提取率产生的影响。按最优提取工艺制备9批五味子乙醇提取物,建立其高效液相色谱(HPLC)指纹图谱。以共有峰峰面积为变量,结合化学计量学方法包括系统聚类分析(HCA)、主成分分析(PCA)法对9批五味子醇提物质量进行分析。结果:确定最佳提取工艺为:采用80%乙醇作为溶剂,料液比为1:10,进行2次回流提取,每次持续1 h。在9批五味子提取物指纹图谱中共有32个共有峰,并成功鉴定出X18(五味子醇甲)、X29(五味子甲素)和X32(五味子乙素),相似度高于0.94。HCA和PCA结果均显示9批样品有聚为四类的趋势,共有峰X1、X2、X3、X13、X17、X19、X21和X30是引起样品间差异的主要化学成分。结论:所优选的提取工艺稳定可行,不同批次五味子提取物的质量一致性较高,但不同样品间仍存在一定差异。

Anti-diabetic Effects of Polysaccharides from Ethanol-insoluble Residue of Schisandra chinensis（Turcz.） Baill on Alloxan-induced Diabetic Mice

The ethanol-insoluble residue of Schisandra generated during lignans industrial production is usually treated as solid waste. However, there is active polysaccharide which could be used in it. In this work, the water-soluble polysaccharides from the ethanol-insoluble residue of Schisandra（ESCP） were obtained and their anti-diabetic effect was evaluated. The results indicate that ESCP could significantly reduce the blood glucose level in alloxan-induced diabetic mice. Moreover, the ESCP could significantly improve the lipid metabolism and increase the content of liver glycogen in alloxan-induced diabetic mice. The results indicate that ESCP could be developed into a potential natural hypoglycemic agent.

五味子种子种苗质量分级标准研究

以五味子23批种子和21批种苗为实验材料,测定五味子种子的净度、千粒重、含水量、生活力、发芽情况等5项指标,种苗的苗长、茎粗、根长、根粗、芽数、侧根数、单株重、每100 g种苗数等8项指标,对种子种苗指标信息进行数据分析,初步制定五味子种子种苗质量分级标准。结果表明,以生活力、发芽率为首要指标,含水量、千粒重、净度为次要指标,将五味子种子分为3个等级标准,一级:净度≥96%,千粒重>29.08 g,含水量>6.75%,生活力>98.22%,发芽率>81.33%;二级:净度≥96%,千粒重25.89~29.08 g,含水量6.52%~6.75%,生活力72.55%~98.22%,发芽率11%~81.33%;三级(不合格):净度<96%,千粒重<25.89 g,含水量<6.52%,生活力<72.55%,发芽率<11%;种苗以苗长、根长、芽数、单株重、每100 g种苗数为分级主要指标,将五味子种苗分为4个等级,一级:苗长>58.177 cm,根长>22.047 cm,芽数>17个,单株重>7.873 g,每100 g种苗数<15棵;二级:苗长>34.803~≤58.177 cm,根长>15.935~≤22.047 cm,芽数>12~≤17个,单株重>3.055~≤7.873 g,每100 g种苗数>15~≤31棵;三级:苗长26.453~34.803 cm,根长14.715~15.935 cm,芽数10~12个,单株重1.442~3.055 g,每100 g种苗数>31~≤70棵;四级(不合格):苗长<26.453 cm,根长<14.715 cm,芽数<10个,单株重<1.442 g,每100 g种苗数>70棵。该研究可为五味子种子种苗质量标准制定提供依据,为其良种选育提供参考。

不同产地五味子种子挥发油中6个成分含量测定及主成分分析

目的:检测不同产地五味子种子挥发油中α-蒎烯、β-蒎烯、月桂烯、柠檬烯、乙酸龙脑酯和榄香烯的含量。方法:采用水蒸气蒸馏法提取挥发油,气相色谱法(GC)测定挥发油中α-蒎烯、β-蒎烯、月桂烯、柠檬烯、乙酸龙脑酯和榄香烯6个成分含量:采用HP-1701石英毛细管色谱柱(30 m×320μm×1μm),氢火焰离子化检测器,进样口温度为230℃,检测器温度为250℃,柱温以60℃为起始温度,以10℃·min–1升温至125℃,保持5 min,再以10℃·min–1升至150℃,保持2 min;载气为氮气,流速为0.9 mL·min–1;分流进样,分流比为10∶1;进样量为1μL。采用SPSSPRO在线数据处理软件,以五味子种子中6个挥发油成分为指标进行综合评价。结果:各产地五味子种子挥发油中6个成分含量差异较大,α-蒎烯为14.3788~231.3942μg·g^(-1),β-蒎烯为6.5591~86.6224μg·g^(-1),月桂烯为11.8734~143.5263μg·g^(-1),柠檬烯为11.2512~113.3705μg·g^(-1),乙酸龙脑酯为122.8318~722.3651μg·g^(-1),榄香烯为11.7830~72.3607μg·g^(-1)。五味子种子挥发油中6个成分含量排序为乙酸龙脑酯>α-蒎烯>月桂烯>柠檬烯>榄香烯>β-蒎烯。结论:采用GC和SPSSPRO软件主成分分析法可以评价五味子的质量,25个产地仅以挥发油为评价指标,黑龙江省伊春市南岔区的五味子质量较好,吉林省敦化市大蒲柴河镇的五味子质量较差。辽宁省的五味子质量整体优于吉林省。

北五味子的繁殖栽培生理及主要活性成分研究进展

五味子[Schisandra chinensis(Turcz.)Baill.]是木兰科五味子属多年生的木质藤本植物。作为一味大宗道地药材,五味子对许多疾病表现出良好的治疗效果和较低的毒性。在现代医学中研究发现五味子有提高人体免疫力、抗抑郁、抗肿瘤、保护受损神经元,提高记忆力等作用,在医疗领域内应用较为广泛。本文对五味子的应用现状、品种选育、扦插、嫁接、组织培养等繁殖技术,花芽分化、光合特性、植物营养元素等栽培生理,黑斑病、白粉病、女贞细卷蛾等常见的病虫害防治,以及五味子甲素、五味子乙素、多糖、挥发油等活性成分药理研究方面进行综述,并对五味子今后的开发利用前景进行展望。

总有机碳检测法在五味子提取清洁验证中的应用

为了证明总有机碳(total organic carbon,TOC)检测法可用于中药材五味子提取设备的清洁验证。采用淋洗法与擦拭法取样,并验证TOC检测法的线性、精密度与准确度、检测限(LOD)与定量限(LOQ)、取样回收率与稳定性,评价总有机碳法用于中成药清洁验证的效果。结果表明,五味子浓度(x)在0.024~0.960 mg/L范围内时,其与TOC值(y)呈良好的线性关系,回归方程为y=218.8x-0.033,r 2=0.999;精密度与准确度的相对标准偏差(RSD)均不超过4%;检测限(LOD)为3.84μg/L,定量限(LOQ)为11.5μg/L;采用两种方式取样,在低、中、高浓度淋洗法平均取样回收率分别为63.91%、81.00%、81.11%,擦拭法平均取样回收率分别为87.50%、83.33%、106.94%;样品溶液分别在0、4、8、24、28、48 h进行TOC检测,RSD为5.21%;应用于生产中减压浓缩罐设备残留量计算,结果符合限度浓度要求。TOC法可用于五味子提取生产清洁验证,可广泛应用于多活性成分的中药制药生产。

响应面法优化酶解-超声双辅助双水相体系制备五味子提取物

在单因素实验的基础上,以五味子乙素得率为响应值,采用响应面法优化了酶解-超声双辅助双水相体系制备五味子提取物的工艺条件,并比较了双水相体系、酶辅助双水相体系、超声辅助双水相体系及酶解-超声双辅助双水相体系等方法对五味子乙素的提取效果。结果表明,酶解-超声双辅助双水相体系制备五味子提取物的最佳条件为:乙醇体积分数27.45%、硫酸铵质量分数25.82%、超声时间56 min、液固比30.98∶1(mL∶g),在此条件下,五味子乙素得率为1.77 mg·g^(-1),与模型预测值(1.76 mg·g^(-1))接近;酶解-超声双辅助双水相体系制备的五味子乙素得率显著高于双水相体系、酶辅助双水相体系及超声辅助双水相体系,说明前处理可有效促进双水相体系对五味子乙素的富集。

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

Objective:Schisandra 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"NanWuweizi"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 ofS.sphenanthera and compared it to that of S.chinensis.Methods:The cp genome of S.sphenanthera was sequenced at the lllumina 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.Results:The cp genome of S.sphenanthera was 146853 bp in length and consisted of a large single copy(LSC)region of 95627 bp,a small single copy(SSC)region of 18292 bp,and a pair of inverted repeats(IR)of 16467 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.Conclusion:We reported the cp genome ofS.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.

北五味子的镇静、催眠作用

目的对北五味子水提取物镇静、催眠药理作用进行初探。方法采用镇静、催眠实验观察北五味子水提取物对小鼠自主活动的影响 ,协同戊巴比妥钠对小鼠的催眠作用。结果北五味子水提取物能明显减少小鼠自主活动次数 ,增加阈下剂量戊巴比妥钠致小鼠睡眠只数 ,延长阈上剂量戊巴比妥钠致小鼠睡眠时间。结论北五味子水提取物有明显的镇静、催眠作用。

Box-Behnken响应面法优选五味子多糖双水相提取工艺

目的:基于Box-Behnken响应面法,采用双水相体系优化五味子多糖的提取工艺。方法:在单因素试验的基础上,选取药液量、K_2HPO_4用量、PEG6000用量为考察因素,以双水相体系的相比、分配系数和萃取效率的总评归一值(OD)为响应值,采用Box-Behnken响应面法筛选提取工艺。结果:最佳的五味子多糖双水相提取参数为:药液量5 m L,K_2HPO_4用量1.0 g,PEG6000用量1.8 g,离心时间9 min。该模型的预测性能良好,验证试验的实测OD值为0.950,与理论预测值的偏差为3.94%。结论:所优选的双水相提取技术操作简便、成本低,可用于五味子多糖的提取。

HPLC法同时测定五味子不同部位中三种木脂素成分含量

目的:采用HPLC法同时测定五味子不同部位中五味子醇甲、五味子甲素和五味子乙素含量。方法:采用Symmetry C18柱,甲醇-乙腈-水(1:1:1)作流动相,流速1ml/min,检测波长250nm。结果五味子醇甲、五味子甲素和五味子乙素含量测定方法的线性关系良好,线性范围和相关系数为0.04～0.2mg/ml(r=0.9999);回收率分别为103.03%、99.83%和101.55%;RSD分别为0.99%、2.15%、1.39%(n=10)。结论:该方法精密度、重现性和分离效果好,分析快速准确,适合于该药材中五味子醇甲、五味子甲素和五味子乙素含量的测定,不同部位三成分均存在一定的差别。

HPLC法同时测定不同产地五味子中8种木脂素类成分

目的测定产自辽宁、河北和黑龙江省的五味子中五味子醇甲、戈米辛J、五味子醇乙、戈米辛G、五味子酯甲、五味子甲素、五味子乙素和五味子丙素8种木脂素成分,为其质量控制提供检测手段。方法五味子用甲醇超声提取,色谱柱为依利特ODS-C18(250 mm×4.6 mm,5μm);流动相为乙腈(A)-水(B),梯度洗脱(0～17 min,A为50%;17～25 min,A为50%～55%;25～30 min,A为55%～75%;30～35 min,A为75%;35～40 min,A为75%～65%;40～45 min,A为65%～50%);体积流量1.0 mL/min;柱温30℃;检测波长217 nm。结果 8种被测木脂素成分分离度良好;各成分质量浓度与峰面积在测定范围内均呈良好的线性关系(r>0.999 5);重复性良好;加样回收率(RSD)(n=9)分别为99.75%(0.38%)、100.69%(2.31%)、100.46%(1.39%)、99.87%(1.15%)、100.22%(1.45%)、100.15%(0.99%)、100.61%(0.25%)和101.31%(1.03%)。结论本实验所建立的HPLC方法稳定可靠、简便可行,可用于五味子中木脂素类成分的同时定量分析。

辽五味子果实挥发油成分的鉴定

目的 对辽五味子的挥发油成分进行鉴定。方法 利用色质联用仪GC/MS产生的双线性数据 ,藉化学计量学发展的二维色谱 /质谱数据解析方法 ,得到了各组分的纯色谱曲线和质谱 ;根据色谱保留时间 ,同时利用解析所得的纯物质质谱在NIST质谱库进行相似检索对各个组分进行定性分析 ,最后利用总体积积分法得到定量结果。结果 在辽五味子干燥果实挥发油中共检出 5 6个组分 ,鉴定了其中 49个化合物 ,占挥发油总量的 98 2 7%。结论此法大大提高了定性定量分析的准确度 ,成功地检测出一些色谱分离不完全和低含量的组分。

响应面法优化超临界CO_2萃取北五味子藤茎油工艺

利用超临界CO2萃取技术,在单因素试验的基础上,采用中心组合响应面法,建立北五味子藤茎挥发油超临界CO2萃取的回归模型。研究结果表明,萃取压力、萃取温度、CO2流速对萃取率的影响显著,萃取压力和CO2流速以及萃取温度和CO2流速的交互效应影响显著,解析矩阵可知,在萃取压力36.32MPa,萃取温度42.27℃、CO2流速17.01L/h,预测最大萃取率为0.432%,验证实验证实该方程有很好的拟合度。该方法具有萃取率高、污染小、节约能源的特点。

北五味子乙素清除自由基及体外抑菌作用的研究

对五味子乙素进行清除自由基及体外抑菌实验。结果表明:五味子乙素对自由基的清除效果明显,其中对羟自由基清除作用大于相同质量浓度的VC,其半数清除质量浓度为0.2mg/mL;对超氧阴离子自由基清除作用小于VC,其半数清除质量浓度为0.24mg/mL。五味子乙素提取液对金黄色葡萄球菌、白色念珠菌、沙门氏菌、大肠杆菌、枯草芽孢杆菌均有一定的抑制作用,其中对白色念珠菌和金黄色葡萄球菌的抑制作用最强,最低抑菌浓度为0.25mg/mL,对根霉、黑曲霉、南洋酵母无明显抑菌活性。