Phenolic derivatives from root bark of Schisandra sphenanthera / 中国中药杂志

This paper aimed to study the chemical constituents from the root bark of Schisandra sphenanthera. Silica, Sephadex LH-20 and RP-HPLC were used to separate and purify the 80% ethanol extract of S. sphenanthera. Eleven compounds were identified by ~1H-NMR, ~(13)C-NMR, ESI-MS, etc., which were 2-[2-hydroxy-5-(3-hydroxypropyl)-3-methoxyphenyl]-propane-1,3-diol(1), threo-7-methoxyguaiacylglycerol(2),4-O-(2-hydroxy-1-hydroxymethylethyl)-dihydroconiferylalcohol(3), morusin(4), sanggenol A(5), sanggenon I(6), sanggenon N(7), leachianone G(8),(+)-catechin(9), epicatechin(10), and 7,4'-dimethoxyisoflavone(11). Among them, compound 1 was a new compound, and compounds 2-9 were isolated from S. sphenanthera for the first time. Compounds 2-11 were subjected to cell viability assay, and the results revealed that compounds 4 and 5 had potential cytotoxicity, and compound 4 also had potential antiviral activity.

Mechanistic understanding of the inhibitory effect of cytochrome P450 3A4 and 3A5 by Wuzhi tablet (Schisandra sphenanthera extract) / 药学学报

Wuzhi tablet (WZ) is a prescribed herbal medicine extracted from Schisandra sphenanthera, which is widely used to protect the liver injury and drug-induced hepatotoxicity in clinical practices. Previous studies showed that WZ significantly increased the blood concentrations of tacrolimus, cyclosporine A, paclitaxel by inhibiting the cytochrome P450 3A (CYP3A)-mediated metabolism. CYP3A4 and CYP3A5 are the most important isoenzymes among the CYP3A subfamily. However, there are some differences in the catalytic and inhibitory activities between CYP3A4 and CYP3A5, which may lead to different risk of drug-drug and herb-drug interactions, and the risks may be further amplified in vivo. Currently, few reports have compared the herbal medicine inhibitory effects between CYP3A4 and CYP3A5 mediated metabolic reactions. Therefore, detailing the inhibitory effect of WZ on CYP3A4 and CYP3A5 will help understand and predict the potential herb-drug interaction. The results showed that WZ inhibited CYP3A4 and CYP3A5 in a NADPH-, time- and concentration- dependent manner. WZ showed more potent inhibition on CYP3A5 than CYP3A4. Cautions warranted when combining WZ with other therapeutic drugs to avoid the potential herb-drug interaction.

Differences between male and female leaves of Schisandra sphenanthera: based on RNA-Seq / 中国中药杂志

Schisandra sphenanthera is dioecious and only the fruits of female plants can be used as medicine and food. It is of great significance for the cultivation and production of S. sphenanthera to explore the differences between male and female plants at the non-flowering stage and develop the identification markers at non-flowering or seedling stage. In this study, the transcriptome of male and female leaves of S. sphenanthera at the non-flowering stage was sequenced by Illumina high-throughput sequencing technology and analyzed based on bioinformatics. A total of 236 682 transcripts were assembled by Trinity software and 171 588 were chosen as unigenes. Finally, 1 525 differentially expressed genes(DEGs) were identified, with 458 up-regulated and 1 067 down-regulated in female lea-ves. The down-regulated genes mainly involve photosynthesis, photosynthesis-antenna protein, carbon fixation in photosynthetic or-ganisms, and other pathways. Real-time quantitative PCR(qPCR) identified two genes between male and female leaves and one of them was a HVA22-like gene related to floral organ development and abscisic acid(ABA). Enzyme linked immunosorbent assay(ELISA) was applied to determine the content of ABA, auxin, gibberellin, and zeatin riboside(ZR) in leaves of S. sphenanthera. The results showed that the content of ABA and ZR in male leaves was significantly higher than that in female leaves. The involvement of down-regulated genes in female leaves in the photosynthesis pathway and the significant differences in the content of endogenous hormones between male and female leaves lay a scientific basis for analyzing the factors affecting sex differentiation of S. sphenanthera.

Study on “Color Discrimination Grading ”Mechanism and Qualitative Identification Model of Schisandra sphe- nanthera and Schisandra chinensis / 中国药房

OBJECTIVE：To investigate the internal mechanism of Schisandra sphenanthera and Schisandra chinensis in determining quality by color （“color discrimination grading ”）of medicinal materials ，and to construct a qualitative identification model based on color quantization value. METHODS ：HPLC method was used to determine the contents of 6 active components from 39 batches of samples. The colorimeter was used to determine 3-color spatial value [lightness value （ΔL＊），red-green value （Δa＊），yellow-blue value （Δb＊）]. SPSS 24.0 statistical software was used to analyze the correlation between the contents of 6 active components and 3-color spatial values. Principal component analysis （PCA）was performed by using SIMCA-P 14.1 software. RESULTS：The linear range of schizandrol A ，schizandrol B ，schisandrin A ，schisandrin B ，schisandrin C ，schisantherin A were 0.204 8-2.560 0，0.049 3-0.616 3，0.098 4- 1.230 0，0.046 3-0.578 8，0.010 6-0.132 0，0.100 0-1.500 0 μg（r＞0.999 0）；RSDs of precision ，stability（12 h）and repeatability tests were all less than 3％. The recoveries were 98.14％-101.53％（RSD＝1.08％， n＝6），97.16％-101.05％（RSD＝1.54％，n＝6），98.29％-101.41％（RSD＝1.29％，n＝6），97.17％-100.36％（RSD＝1.20％，n＝ 6），97.32％-102.43％（RSD＝1.77％，n＝6）and 98.02％-100.40％（RSD＝0.84％，n＝6），respectively. Among 39 batches of components were 3.25-7.39，0.96-1.98，0.46-4.74，1.62-2.60， 0.06-0.58，0.48-6.11 mg/g，respectively. Average S. chinensis was － 80.79-－ 70.54， average Δ a ＊ was qq.com # 通 2.54-5.34，average Δb＊ was 5.20-12.83，average ΔE＊ was 71.13-81.23；average ΔL＊ of S. sphe nanthera was －75.90- －69.16，average Δa＊ was 3.77-7.82，average Δb＊ was 8.59-17.23，average ΔE＊ was 69.99-77.92. The results of relationship analysis showed that the contents of schizandrol A ，schizandrol B ，schisandrin A ，schisandrin B and schisantherin A were significantly correlated with ΔL＊，Δa＊，ΔE＊（P＜0.01），with no significant correlation with Δb＊（P＞0.05）. There was a negative correlation of the content of schisandrin C with ΔL＊ and Δa＊（P＜0.05），and there was no significant correlation with Δb＊ and ΔE＊ （P＞0.05）. Results of PCA showed that accumulative variance contribution rate of primary 2 main components was 89.8％，and S. sphenanthera and S. chinensis could be identified significantly. CONCLUSIONS ：The content of schizandrol A in S. chinensis is high relatively ，and content of schisantherin A in S. sphenanthera is high relatively. Schizandrol A ，schizandrol B and schisandrin B were not detected in S. sphenanthera . The 3-color spatial value of S. sphenanthera and S. chinensis are different ，that is ，the brightness of S. chinensis is small and the color is slant black ，while the color of S. sphenanthera is slant red and yellow. The contents of active components of S. sphenanthera and S. chinensis is related to the surface 3-color spatial values ，that is ，the darker the color is ，the weaker the red degree is ，and the higher the contents of schizandrol A ，schizandrol B ，schisandrin B and schisandrin C are ；the brighter the surface color is ，the stronger the red degree is ，and the higher the contents of schisandrin A and schisantherin A are. The established content determination method is precise and stable ，and can be used for the content determination of S. sphenanthera and S. chinensis . The color qualitative identification model can be used for the identification of S. sphenanthera and S. chinensis .

Comparing chloroplast genomes of traditional Chinese herbs Schisandra sphenanthera and S. chinensis / 中草药·英文版

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 “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. Methods: The 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. Results: The 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. Conclusion: We 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.

Establishment of HPLC Fingerprint of Schisandra sphenanthera and S. chinensis and Analysis of Chemical Pat- tern Recognition / 中国药房

OBJECTIVE：To establish HPLC fingerprint of Schisandra sph enanthera and S. chinensis，and to analyze chemical pattern recognition. METHODS ：HPLC method was adopted. Using schizandrin A as reference ，HPLC fingerprints of 10 batches of S. sphenanthera and S. chinensis （N1-N10，S1-S10） were drawn. Similarity Evaluation System of TCM Chromatographic Fingerprint（2012 edition）was adopted for similarity evaluation to determine the common peaks. SPSS 20.0 and SIMCA 14.1 software were used for HCA ，unsupervised madel of PCA ，supervised model of OPLS-DA. Using variable importance projection （VIP）value greater than 1 as the standard ，the differential markers that affected the quality of S. sphenanthera and S. chinensis were screened. RESULTS ：S. sphenanthera and S. chinensis were identified 32 and 33 common peaks ，respectively. The similarity of 10 batches of S. sphenanthera and 10 batches of S. chinensis were all higher than 0.9，and the similarity of S. sphenanthera and S. chinensis was 0.05. A total of 19 characteristics peaks were identified ，among which five common peaks were identified as schisandraol A ，schisandraol B ，schisantherin A ，schizandrin A and schisandrin B by reference. HCA results showed that N 1-N10 were clustered into one category ，and S 1-S10 were clustered into one category ，of which N 1，N3，N8，and N 9 were clustered into one category ，and the rest were clustered into one category ；S1，S3，S6，and S 9 were grouped together ，and the rest were grouped together. The results unsupervised model of PCA showed that the cumulative variance contribution rate of the first two principal component factors was 87.20％. Supervised model of OPLS-DA showed that schizandrin A ，schisandraol A ，schisantherin A and schisandrin B were the differential markers that affected 、the quality of S. sphenanthera and S. chinensis （VIPs were 2.29，2.24，1.73，1.48，respectively）. CONCLUSIONS ：The established fingerprint is accurate ，scientific，simple and easy to use ，combined with multivariate statistical analysis can be 话：0395-3356116。E-mail：wangrui56116@163.com used to evaluate the quality of S. sphenantherae and S. chinensis. The components of S. sphenanthera and S. chinensis were different ，schisanolrin A is differential marker.

Classification analysis for Schisandra sphenanthera during different grades of maturity by chemical pattern recognition approach / 中草药

Objective To establish the simultaneous determination method of six lignans from Schisandra sphenanthera and classify the different maturity of Schisandra sphenanthera via chemical pattern recognition approach. Methods The simultaneous determination method was established by UPLC technology, the samples were performed by Acquity UPLC BEH C18 (50 mm × 2.1 mm, 1.7 μm) and eluted with acetonitrile and 0.1% formic acid, the flow rate was 0.4 mL/min, column temperature was 35 ℃, and the detection wavelength was 254 nm. The collected data were analyzed and classified by principal component analysis (PCA) and hierarchical cluster analysis (HCA) that the Schisantherin was selected as the monitoring component. Results Six lignans were in good linear relationship with the linear range 1.257-156.250 μg/mL (r < 0.999); Moreover, the RSD values of the repeatability, stability, and precision test were all less than 3%. According to the variation of main components in different grades of maturity, six different grades of maturity of S. sphenanthera were divided into three groups: 60%-mature sample, 70%-mature sample, and mature sample. Conclusion Cluster analysis based on UPLC technology and PCA joint analysis method are simple, rapid, sensitive, and accurate, which provides technical support and scientific basis for the selection of quality control and harvest time.

Process optimization and component analysis of extracting seed oil from Schisandra sphenanthera by supercritical CO2 / 国际药学研究杂志

Objective To optimize the extraction conditions of Schisandra sphenanthera(SS)seed oil by supercritical CO2ex?traction and identify its components by GC-MS.Methods SS seed oil was used as tested material,response surface methodology was used to optimize the process of supercritical CO2extraction,and GS-MS method was used to analyse SS seed oil composition.Results A model of an equation was established,which could be used to optimize the process parameters of supercritical CO2extraction of SS seed oil. The optimum extraction parameters were as follows:the extraction pressure was 33 MPa,the extraction temperature was 53℃,the extraction time was 90 min,the CO2flow rate was 21.40 ml/min.In this condition,the extraction rate of SS seed oil was 7.97%.The SS seed oil was analyzed by GC-MS,and 23 compounds were identified.In these compounds,(1α,4a.β,8a.α)-1,2,3, 4,4a,5,6,8a-octahydro-7-methyl-4-methylene-1-(1-methylethenyl)-naphthalene,(-)-1,7-dimethyl-7-(4-methyl-3-pentenyl)-tricyc?lo[2.2.1.0(2,6)]heptane and(R)-2,4a,5,6,7,8-octahydro-3,5,5,9-tetramethyl-1H-(σ-phenyl)cycloheptene had the content of more than 10% and the contents were 27.78%,14.77% and 13.12% respectively.Conclusion This process has high extraction rate, fast speed and simple operation,and can be used for the extraction of SS seed oil.

Chemical constituents from canes of Schisandra sphenanthera / 中草药

Objective: To investigate the chemical constituents from the canes of Schisandra sphenanthera. Methods: The compounds were isolated and purified by chromatography on silica gel. Their structures were elucidated on the basis of spectroscopic methods, including MS, 1D, and 2D NMR spectral techniques. Results: Thirteen compounds were isolated from 95% ethanol extract in the canes of S. sphenanthera, including six lignans, six triterpenes, and one steroid and their structures were identified as sphendilactone (1), kadsulactone (2), schisanlactone E (3), 12-β-hydroxycoccinic acid (4), schizandronic acid (5), schisanbilactone A (6), β-sitosterol (7), meso-dihydroguaiaretic acid (8), deoxychizandrin (9), schizantherin A (10), schisanhenol B (11), γ-schizandrin (12), and R-(+)-angeloygomisin M1 (13). Conclusion: Among them, one new triterpene is named as sphendilactone (1). Schisanhenol B, deoxychizandrin, R-(+)-angeloygomisin M1, kadsulactone, schisanlactone E, 12β-hydroxycoccinic acid, and schisanbilactone A are isolated from this plant for the first time.

Rapid identification on chemical constituents in fruits of Schisandra sphenanthera and Schisandra chinensis and discrimination markers by UPLC-Q-TOF-MSE combined with OPLS-DA / 中草药

Objective: To establish a method to fast identify the chemical constituents in the fruits of Schisandra sphenanthera and Schisandra chinensis by ultra performance liquid chromatography tandem with time-of-fight mass spectrometry with MSE data-acquistion mode (UFLC-Q-TOF-MSE) and provide the basis for high throughput characterization and distinction of the two two samples of Schisandra Michx by analyzing the remarkably different chemical components in the fruits between S. sphenanthera and S. chinensis. Methods: To rapidly identify the main chemical constituents in the fruits between S. sphenanthera and S. chinensis by using time-dependent scan mode, and combine with orthogonal partial least squares discriminant analysis (OPLS-DA) method to acquire significantly different constituents in the two samples of Schisandra Michx. Results: A total of 33 compounds were identified including 27 dibenzocyclooctene lignans. The differences of chemical compounds between the two sample groups could be obviously observed through the method of OPLS-DA in positive mode. There were 15 chemical ingredients of lignanes with significant differences identified in the two sample groups by comparison with retention time and mass spectra. Conclusion: The present study provides the basis for rapidly qualitative analysis of constituents and quantity control of the fruits in S. sphenanthera and S. chinensis.

Pharmacognostical identification of Chinese crude drug Fructus Schisandrae Sphenantherae / 中草药

Object To provide a basis for the identification of the crude drug of Fructus Schisandrae Sphenantherae Methods Morpholgical characters and microstructural features of the seed surface of Schisandra sphenanthera Rehd. et Wils , S. rubriflora (Franch ) Rehd. et Wils., S. viridis A. C. Smith, and S. henryi Clarke were observed under light microscope and scanning electron microscope. Identification of them was also completed by TLC qualitative analysis for deoxyschisandrin and schisantherin A Results The fruits of S. sphenanthera, S. rubriflora, S. viridis and S. henryi could be classified to three types with the characters of micromorphological features of the seed surface. The results of TLC showed that deoxyschisandrin and schisantherin A were present in the fruits of S. sphenanthera from Pingli, Shaanxi, Luanchuan, Henan, Yangcheng, Shanxi, S. rubriflora and S.viridis, Hengshan Hunan Provinces The fruit of S. henryi contained a little quantity of schisantherin A, but the fruits of S. sphenanthera from Liuba Shaanxi and Xiaolongshan Gansu Provinces did not have deoxyschisandrin and schisantherin A. Conclusion The fruits of S. sphenanthera from different geographical origin, S. rubriflora, S. viridis and S. henryi can be identified by the characters of micromorphological featrues of the seed surface and the TLC qualitative analysis for deoxyschisandrin and schisantherin A