Characteristics and Antioxidant Activity of Lignans in Schisandra chinensis and Schisandra sphenanthera from Different Locations

Twenty Schisandra samples were collected from different locations. Contents of 7 lignans in the samples were determined and analyzed by HPLC method coupled with hierarchical clustering analysis (HCA) and principal component analysis (PCA), and the antioxidant capacity of Schisandra from the different locations was evaluated by reducing power, ferric thiocyanate (FTC) and 2,2′‐diphenyl‐1‐picrylhydrazyl (DPPH) assay. The results showed that there was a significant difference in the content of lignans between Schisandra chinensis and Schisandra sphenanthera. The Schisandra sphenanthera samples in the southwest of China were significantly different from those from the other locations. The antioxidant capacity of Schisandra chinensis was significantly superior to that of Schisandra sphenanthera, and the main antioxidant components were schisandrol A, schisandrol B and schisandrin B based on the result of discrimination analyses. The differences in the chemical composition and antioxidant activity of lignans in Schisandra chinensis and Schisandra sphenanthera from the different locations were investigated in this study, which may provide an experimental basis for the quality control of Schisandra.     

Nortriterpenoids and lignans from Schisandra sphenanthera

Nortriterpenoids, sphenadilactone C (1) and sphenasin A (2), together with four known lignans (3-6), were isolated from the leaves and stems of Schisandra sphenanthera. Their structures were elucidated by extensive analysis of 1D and 2D NMR spectroscopic data and compound 2 was further confirmed by single-crystal X-ray diffraction. Compound 1 features a partial enol moiety and an acetamide group in its structure. In addition, compounds 1, 3-6 showed weak anti-HIV-1 activity with EC₅₀ values in the range of 15.5-29.5 μg/mL.     

Anti-liver fibrotic lignans from the fruits of Schisandra arisanensis and Schisandra sphenanthera

Three new polyoxygenated C₁₈-dibenzocyclooctadiene lignans, arisanschinins M and N (1 and 2) and schisphenin A (3), together with eight related metabolites (4–11), were isolated from the fruits of Schisandra arisanensis and Schisandra sphenanthera, respectively. The structures of 1–3 were elucidated on the basis of extensive spectroscopic and 2D NMR (HSQC, HMBC, and NOESY) analyses. The configuration of the biphenyl moiety in the octadiene ring was determined by circular dichroism (CD). Compound 1 possessed an unprecedented 3-(1-hydroxypropan-2-yl)-3-methyl-1,4-dioxo-2-one lactonide ring system attaching at C-6/C-14. Pharmacological studies revealed that compounds 3, 4, 6, 7, and 10 exhibited significant anti-hepatic fibrosis activity, while 9 and 11 showed cytotoxicity against HSC-T6 cells. The biogenetic pathway for compound 1 was also proposed.     

Seven New Sesquiterpenoids from the Fruits of Schisandra sphenanthera

Fractionation of the EtOH extract from the fruits of Schisandra sphenanthera resulted in the isolation of seven new sesquiterpenoids, 1 – 7 , in addition to the known metabolites 8 – 23 . Among them, schiscupatetralin A ( 1 ) possesses an unprecedented structure with a C? C bond between cuparenol and tetralin. The isolated new compounds were evaluated for their anti‐HSV‐1 and anti‐inflammatory activities. The results revealed that compound 4 exhibited anti‐HSV‐1 activity, while compound 6 showed a significant anti‐inflammatory activity.     

Lignans and Terpenoids from the Leaves of Schisandra chinensis

Fifteen constituents, including one new lignan (schisandroside E) and one new terpenoid (schisandenoid A) as well as nine known lignans and four known terpenoids, were isolated from Schisandra chinensis leaves. The structures of schisandroside E and schisandenoid A were established by entirely meticulous spectroscopic analysis (NMR, MS, CD, IR and UV). All compounds were tested for cytotoxicity against MGC‐803, Caco‐2 and Ishikawa cell lines. Some compounds showed strong cytotoxicity against these three cancer cell lines with IC₅₀<1 μm .     

不同产地华中五味子叶表皮结构和导管分子的解剖学特征及其与环境因子的关系

在光学显微镜下观察了不同产地华中五味子(Schisandra sphenanthera Rehd.et Wils．)叶表皮结构和导管分子特征,应用多元回归方法对不同产地华中五味子的叶表皮特征及导管分子特征与环境因子的关系进行了分析。观察的指标有叶表皮特征(气孔器密度、气孔器指数、气孔器长宽、气孔器面积、气孔极区角质加厚、气孔器类型、表皮细胞垂周壁形状、分泌细胞密度及叶表面角质条纹)和导管分子特征(导管分子长度、直径、长／直径比值及穿孔板类型)。观察结果：上表皮有极少量气孔器和分泌细胞分布。在下表皮,气孔器类型为平列型和侧列型,气孔器指数为15．05％-22．53％,气孔器长59．2—74．2μm,气孔器宽37．9—46．8μm,分泌细胞密度为1．45／mm^2-3．99／mm^2。导管分子以具缘纹孔导管为主,穿孔板包含单穿孔和梯状穿孔板。导管分子长957．8—1270．2μm,导管分子直径85．0—136．7μm。随着产地的不同,华中五味子叶下表皮结构和导管分子的解剖特征有明显的差异。分析结果显示：年均气温与气孔器面积和气孔器长度,年降水量与气孔器宽度以及相对湿度与分泌细胞密度之间关系密切。随着年平均气温的升高,气孔器面积、气孔器长度呈减少的趋势,R^2值分别为0．74、0．71。随着年降水量的增加,气孔器宽呈减小的趋势,R^2值为0．64;导管分子长／直径比值增加,R^2值为0．46。相对湿度与分泌细胞密度呈正相关,R^2为0．63。     

Phytochemical studies and biological activity of three Chinese Schisandra species (Schisandra sphenanthera,Schisandra henryi and Schisandra rubriflora): current findings and future applications

Phytochemistry Reviews - Schisandra chinensis (Chinese magnolia vine) is the best-known representative of the genus Schisandra, widely used in the official East-Asian, North American and European...     

New Thymoquinol Glycosides and Neuroprotective Dibenzocyclooctane Lignans from the Rattan Stems of Schisandra chinensis

Three new lignans ( 1 – 3 ), together with four new thymoquinol glycosides ( 4 – 7 ), were isolated from 70%‐EtOH extract of the rattan stems of Schisandra chinensis. The structures of 1 – 7 were elucidated by detailed spectroscopic analyses, and these new compounds were identified as pinobatol‐9‐O‐β‐d ‐glucopyranoside ( 1 ), 1,2,13,14‐tetramethoxydibenzocyclooctadiene 3,12‐O‐β‐d ‐diglucopyranoside ( 2 ), 3,7‐dihydroxy‐1,2,13,14‐tetramethoxydibenzocyclooctadiene 12‐O‐β‐d ‐glucopyranoside ( 3 ), thymoquinol 2‐O‐β‐d ‐apiofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 4 ), thymoquinol 2‐O‐α‐d ‐arabinofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 5 ), thymoquinol 5‐O‐β‐d ‐apiofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 6 ), and thymoquinol 5‐O‐α‐d ‐arabinofuranosyl‐(1→6)‐β‐d ‐glucopyranoside ( 7 ). The neuroprotective activity of 1 – 7 was evaluated on PC12 cells with neurotoxicity induced by amyloid‐beta 1 – 42 (Aβ_{1 – 42}). Compounds 2 and 3 showed protecting activity against Aβ‐induced toxicity in PC12 cells.     

华中五味子ISSR-PCR反应体系优化及引物筛选

系统研究了华中五味子ISSR PCR反应体系中的主要影响因子,确立华中五味子ISSR-PCR最适反应体系,并筛选出12条有效引物。单因子实验结果显示,华中五味子ISSR-PCR反应体系中各主要成分的适宜浓度范围为,Mg²⁺ 1.50~3.50 mmol·L^-1,dNTPs 0.10~0.35 mmol·L^-1,引物0.25~0.60 μmol·L^-1,Taq酶0.50~1.50 U。4因子3水平正交实验确立了最适反应体系,即20 μL体系中包含2.50 mmol·L^-1 Mg²⁺、0.20 mmol·L^-1 dNTPs、0.25 μmol·L^-1引物、1.50 U Taq酶、60 ng DNA模板、2.50 μL 10×PCR Buffer。本研究为华中五味子种质资源的评估及遗传多样性分析奠定基础。     

药用植物华中五味子的种群遗传多样性及遗传结构

华中五味子（Schisandra sphenanthera）是著名的药用植物,具有悠久的药用历史和巨大的开发潜力。为了有效评估、利用和保护华中五味子资源,应用自主开发的9对SSR引物研究了华中五味子自然种群的遗传多样性与遗传结构。结果表明：在10个采样种群中,共检测到58个等位基因,平均预期杂合度H_E为0.528,平均观察杂合度H_O为0.519,较大的连续种群保持了较高的遗传多样性,而小种群的遗传多样性则相对较低;华中五味子总体表现为显著的杂合子缺失,内繁育系数F_IS为0.042;种群间总的遗传分化系数F_ST为0.108,两两种群间分化显著;贝叶斯聚类结果把10个采样种群按遗传组成分为江南组和江北组2组,长江所形成的特殊地理屏障对华中五味子江南和江北地区间较高的遗传分化造成了影响。     

Intercarpellary growth of pollen tubes in the extragynoecial compitum and its contribution to fruit set in an apocarpous species, Schisandra sphenanthera (Schisandraceae)

? Premise of the study: Apocarpous plants possess carpels that are separated in the gynoecium. Extragynoecial compita, commonly occurring in basal angiosperms, have been proposed to have the potential to increase offspring quantity in apocarpous species through the intercarpellary growth of pollen tubes. To date, the impact of an extragynoecial compitum on fruit or seed set has not been studied in any species. This study investigated the pollen tube pathway between adjacent carpels and its contribution to fruit set in Schisandra sphenanthera. ? Methods: We investigated the fruit set ratio in the field and collected hundreds of gynoecia at their full flowering stage. Pollinated carpel ratio and pollen tube pathway observations were performed using fluorescence optics. ? Key results: Pollen grains germinated and tubes extended along the pseudostyle surface. Some of them turned and entered the ovules at the end of the stigmatic crest, whereas others subsequently grew into neighboring carpels through promontory connections located at the base of the unfused carpels. No tubes were found growing on the surface of the receptacle. More than 24 carpels could be fertilized by pollen tubes from one carpel through hand pollination. The pollinated carpel ratio was significantly lower than the fruit set ratio under natural conditions. ? Conclusions: Pollen tubes from one carpel can easily cross in the extragynoecial compitum between the adjacent carpels of S. sphenanthera, and this intercarpellary growth of pollen tubes can significantly increase the fruit set of apocarpous species, at least in S. sphenanthera.     

安徽黄山华中五味子次生木质部的生态解剖

对安徽黄山海拔420～1840m的华中五味子茎的次生木质部进行了生态解剖学研究。结果表明,茎的次生木质部为散孔材,导管分子长741.7～1025.2μm,直径为152.4～191.9μm;导管频率60.6～70.2mm-2。纤维管胞长925.2～1046.3μm;木射线类型为异形ⅡA和ⅡB,单列射线高682.1～778.4μm,多列射线高度为1093～1208μm,多列射线宽63.6～92.6μm,射线频率9.2～12.8mm-1。次生木质部解剖特征随异质生境而表现出一定的可塑性,其中,多列射线宽、射线频率、导管分子长度、导管直径等性状的可塑性较大。多重回归分析表明,水分和温度是影响华中五味子次生木质部解剖特征的主导因子。随着空气相对湿度的增加,导管分子长度和直径均增大,射线频率减小。随着降雨量的增加,纤维长度增加。随着最冷月温度的增加,导管频率增加,多列射线宽度减小。随着年较差的增加,单列射线和多列射线高度均减小。     

华中五味子（五味子科）雄花和雌花的形态发生

通过扫描电镜观察了中国特有种华中五味子（ＳｃｈｉｓａｎｄｒａｓｐｈｅｎａｎｔｈｅｒａＲｅｈｄｅｔ．Ｗｉｌｓ）,雄花和雌花的形态发生过程,雄花：花被片和雄蕊连续向顶发生,未见有雌性结构的分化,花药的分化先花丝,雄蕊始终螺旋状排列在柱状花托上,雌花：花被片和心皮也为２／５序列连续地螺旋状向顶发生,未见有雄性结构的分化,心皮原基近轴面基部边缘的活动不明显,心皮为对折型：胚珠原基在心皮近轴面上近边缘处发生     

Ontogeny of Staminate and Carpellate Flowers of Schisandra sphenanthera(Schisandraceae)

The ontogenetic process of the staminate and carpellate flowers of Schisandra sphenanthera Rehd. et Wils., an endemic species to China, was observed for the first time under the scanning electron microscope (SEM). In the staminate flowers, the perianth units and stamens were initiated acropetally in a continuous fasion with 2/5 spiral phyllotaxis, while no female structures were formed. Anthers were differentiated prior to the filaments formation. Throughout all the stages were the stamens arranged spirally on a columniform receptacle. In the carpellate flowers, the initiation sequence of the perianth units and carpels were similar to that of the staminate flowers. In contrast, no male structures were formed. Shortly after initiation, the carpel primordia began their marginal growth besides the apical growth and then appresses were formed on the adaxial surfaces of the primordia. However the lower margins of these appresses were inconspicuous, resulting in conduplicate carpels. Two ovules were developed on the inner surface near either lateral margins of the carpel, shaping laminar placentae. Compared with S. glabra (Brickell) Rehd., a related American species, the evolutionary trend of phyllotaxis of androecia is considered that stamens may change from spiral to approximately whorled arrangement, accompanying with the change of receptacle from a column to a flattened shield. It was also suggested that the stamens being numerous and uncertain in number become certain and decrease in number to 5 (4－7). Sterile stamens are observed and the unisexual nature of the flowers is discussed. Two types of carpel primordia are categorized, corresponding to two types of carpels, namely, ascidiate and conduplicate carpels, respectively.     

Isolation and characterization of polymorphic microsatellite loci in a traditional Chinese medicinal plant, Schisandra sphenanthera

Schisandra sphenanthera (Schisandraceae) is a traditional Chinese medicinal plant. In this study, nine polymorphic microsatellite loci were developed and characterized for the species. A genomic DNA enrichment protocol was used to isolate microsatellite loci and polymorphism was explored using 36 individuals sampled from a natural population. The observed number of alleles ranged from 2 to 10 with an average of 5.2. Expected and observed heterozygosity ranged from 0.30 to 0.82 and 0.33 to 0.97, respectively. These microsatellites have been directly applied to both population and conservation genetic studies of S. sphenanthera.     

华中五味子抗氧化和细胞毒活性研究

我们研究了华中五味子的根、茎、叶、果实乙醇提取物对脂性自由基DPPH及超氧阴离子的清除作用,同时观察了四种不同提取物在体外对肿瘤细胞株的细胞毒作用。我们发现,华中五味子根的乙醇提取物清除自由基活性和对肿瘤细胞杀伤作用比茎、叶和果实的提取物都强,研究结果提示华中五味子的根可以作为抗氧化剂的新来源。     

华中五味子干燥材料DNA提取方法的研究

目的：建立一套适合从华中五味子硅胶干燥的幼芽中提取高质量DNA的方法,为进一步开展分子生物学研究奠定基础。方法：比较SDS法、SIX-CTAB法、SDS-CTAB法以及经过作者改进的CTAB法,运用紫外光谱和电泳分析方法测定所得DNA样品的纯度与完整性。结果：四种方法中改进CTAB法所得DNA纯度最高,完整性好,其A260/A280值为1．81,A260/A230值为2．13,经稳定性试验证明该方法稳定。结论：改进CTAB法是一套适合从华中五味子硅胶干燥的幼芽中提取高质量DNA的方法。     

华中五味子叶表型可塑性及环境因子对叶表型的影响

表型可塑性是指同一基因型的生物个体在不同环境条件下产生不同表型的特性,这一特性促使物种适应异质环境,具有更宽的生态幅。居群和物种水平广义的表型可塑性研究可揭示植物在自然环境中的生态意义并丰富对植物种群分布的了解。华中五味子(Schisandra sphenanthera)是一种资源严重减少的常用野生药材物种,对其进行研究和保护非常必要。叶是华中五味子主要的光合作用器官,随环境改变叶表型有很大变化,因此从居群水平上对华中五味子叶表型可塑性进行分析,揭示叶表型可塑性大小,探究环境因子对叶表型的影响,可为华中五味子的保护提供依据。该研究运用GIS技术从较大尺度(9省市26县27个居群)对华中五味子叶表型可塑性进行分析,并用表型可塑性指数和变异系数2种方法对叶表型可塑性进行评价,运用隶属函数和最大熵模型将3类(土壤、气候和地形)13个环境因子对叶表型的影响进行分析。结果表明,华中五味子7个叶表型(叶长、叶宽、叶柄长、叶面积、叶形指数、叶齿数和二级叶脉数)均存在可塑性,其中叶面积可塑性最大,叶形指数可塑性最小。3类环境因子中对叶表型影响最大的为土壤因子,气候因子次之,最小为地形因子。权重分析显示,土壤全钾含量对叶长和叶面积影响最大,有机碳含量、海拔、土壤全磷含量、坡度和年降水量分别对叶宽、叶柄长、叶形指数、叶齿数和二级叶脉数影响最大。研究结果表明,对多个叶表型有影响的5个共有重要因子为土壤全钾含量、全磷含量、有机碳含量、年降水量和坡度,它们与对应叶表型间的隶属函数多为高斯型,具有最优值,可通过人工控制达到最优值来促进其生长。该结果为华中五味子野生抚育和迁地种植提供理论参考。     

Expression of unselected adenovirus genes in human cells co-transformed with the HSV-1 tk gene and adenovirus 2 DNA

We have introduced adenovirus 2 genes into high molecular weight DNA of permissive human cells by co-transformation of tk- human 143 cells with Ad2 restriction enzyme fragments and a cloned Bam HI fragment that carries the HSV-1 thymidine kinase gene. Tk+ cells were isolated after selection and maintenance in HAT medium. Several co-transformed lines are able to complement the growth of Ad5 dl312 (delta 1.2--3.7) and Ad5 dl434 (delta 2.6--8.7), deletion mutants that lack sequences from the left end of the viral genome. The amount and arrangement of viral sequences in the co-transformed cell lines have been analyzed by restriction endonuclease digestion and filter hybridization. Most of the cell lines contain a single insertion of the HSV-1 tk fragment and a single insert of adenoviral DNA. However, one line (B1) contains at least four different insertions, two of which are present in multiple copies. The adenoviral DNA in all cell lines is composed of sequences from the left end of the genome and extends for varying lengths in different lines. Two cell lines that complement deletion mutants efficiently synthesize both early region 1a and 1b mRNAs. The B1 line synthesizes low levels of 1a mRNA, higher levels of 1b mRNA and a unique mRNA that maps to the right of the 1b gene family. When grown continuously in HAT medium, some cell lines are quite stable while others are fairly unstable. Some tk+ subclones support the growth of viral mutants as well as the parental line while others give reduced levels of complementation. For all tk+ subclones examined, the alteration or reduction in viral gene expression is independent of changes in the pattern of integration of viral DNA.