Anxiolytic-like effect of Suanzaoren-Wuweizi herb-pair and evidence for the involvement of the monoaminergic system in mice based on network pharmacology.

BACKGROUND: Suanzaoren-Wuweizi herb-pair (SWHP), composed of Zizyphi Spinosi Semen (Suanzaoren in Chinese) and Schisandrae Chinensis Fructus (Wuweizi in Chinese), is a traditional herbal formula that has been extensively used for the treatment of insomnia. The study aimed to explore the targets and signal pathways of Suanzaoren-Wuweizi (S-W) in the treatment of anxiety by network pharmacology, and to verify the pharmacodynamics and key targets of SWHP in mice. METHODS: The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) as well as literature mining were used to obtain the main chemical ingredients of Suanzaoren and Wuweizi. The SwissTargetPrediction platform was used to predict drug-related targets. The GeneCards, TTD, DisGeNET and OMIM databases were used to obtain potential targets for the treatment of anxiety with the chemical components of S-W. Drug-disease intersection genes were selected, and a protein-protein interaction (PPI) network was constructed using STRING. The core targets of S-W in the treatment of anxiety were selected according to the topological parameters, and GO functional enrichment as well as KEGG pathways enrichment analyses were performed for potential targets. The relationship network of the "drug-active ingredient-disease-target-pathway" was constructed through Cytoscape 3.8.0. The pharmacodynamics of SWHP in the treatment of anxiety was evaluated by the elevated plus maze (EPM), the light/dark box test (LDB) and the open field test (OFT). The mechanisms were examined by measuring monoamine neurotransmitters in brain of mice. RESULTS: The results showed that there were 13 active ingredients for the treatment of anxiety in the network. This includes sanjoinenine, swertisin, daucosterol, schizandrer B, wuweizisu C and gomisin-A. Additionally, there were 148 targets, such as AKT1, TNF, SLC6A4, SLC6A3, EGFR, ESR1, HSP90AA1, CCND1, and DRD2, mainly involved in neuroactive ligand-receptor interactions, the Serotonergic synapse pathway and the cAMP signaling pathway. After 1 week of treatment, SWHP (2 and 3 g/kg) induced a significant increase on the percentage of entries into and time spent on the open arms of the EPM. In the LDB test, SWHP exerted anxiolytic-like effect at 2 g/kg. In the open-field test, SWHP (2 g/kg) increased the number of central entries and time spent in central areas. The levels of brain monoamines (5-HT and DA) and their metabolites (5-HIAA, DOPAC) were decreased after SWHP treatment. CONCLUSIONS: The anti-anxiety effect of SWHP may be mediated by regulating 5-HT, DA and other signaling pathways. These findings demonstrated that SWHP produced an anxiolytic-like effect and the mechanism of action involves the serotonergic and dopaminergic systems, although underlying mechanism remains to be further elucidated.

A comprehensive review of ethnopharmacology, phytochemistry, pharmacology, and pharmacokinetics of Schisandra chinensis (Turcz.) Baill. and Schisandra sphenanthera Rehd. et Wils.

ETHNOPHARMACOLOGICAL RELEVANCE: Schisandra chinensis (called bei-wuweizi in Chinese, S. chinensis) and Schisandra sphenanthera (called nan-wuweizi in Chinese, S. sphenanthera) are two highly similar plants in the Magnoliaceae family. Their dried ripe fruits are commonly used as traditional Chinese medicine in the treatment of coughs, palpitation, spermatorrhea, and insomnia. They also are traditionally used as tonics in Russia, Japan, and Korea. AIM OF THE REVIEW: S. chinensis and S. sphenanthera are similar in appearance, traditional applications, ingredient compositions, and therapeutic effects. This review, therefore, aims to provide a systematic insight into the botanical background, ethnopharmacology, phytochemistry, pharmacology, pharmacokinetics, quality control, and toxicology of S. chinensis and S. sphenanthera, and to explore and present the similarities and differences between S. chinensis and S. sphenanthera. MATERIALS AND METHODS: A comprehensive literature search regarding S. chinensis and S. sphenanthera was collected by using electronic databases including PubMed, SciFinder, Science Direct, Web of Science, CNKI, and the online ethnobotanical database. RESULTS: In the 2020 Edition of Chinese Pharmacopoeia (ChP), there were 100 prescriptions containing S. chinensis, while only 11 contained S. sphenanthera. Totally, 306 and 238 compounds have been isolated and identified from S. chinensis and S. sphenanthera, respectively. Among these compounds, lignans, triterpenoids, essential oils, phenolic acid, flavonoids, phytosterols are the major composition. Through investigation of pharmacological activities, S. chinensis and S. sphenanthera have similar therapeutic effects including hepatoprotection, neuroprotection, cardioprotection, anticancer, antioxidation, anti-inflammation, and hypoglycemic effect. Besides, S. chinensis turns out to have more effects including reproductive regulation and immunomodulatory, antimicrobial, antitussive and antiasthmatic, anti-fatigue, antiarthritic, and bone remodeling effects. Both S. chinensis and S. sphenanthera have inhibitory effects on CYP3A and P-gp, which can mediate metabolism or efflux of substrates, and therefore interact with many drugs. CONCLUSIONS: S. chinensis and S. sphenanthera have great similarities. Dibenzocyclooctadiene lignans are regarded to contribute to most of the bioactivities. Schisandrin A-C, schisandrol A-B, and schisantherin A, existing in both S. chinensis and S. sphenanthera but differing in the amount, are the main active components, which may contribute to the similarities and differences. Study corresponding to the traditional use is needed to reveal the deep connotation of the use of S. chinensis and S. sphenanthera as traditional Chinese medicine. In addition, a joint study of S. chinensis and S. sphenanthera can better show the difference between them, which can provide a reference for clinical application. It is worth mentioning that the inhibition of S. chinensis and S. sphenanthera on CYP3A and P-gp may lead to undesirable drug-drug interactions.

Rapid analysis of nine lignans in Schisandra chinensis by supercritical fluid chromatography using diode array and mass spectrometric detection.

Lignans are the bioactive constituents in Schisandra chinensis fruits. For the first time major representatives could directly be determined in plant extracts by using Supercritical Fluid Chromatography. Based on nine commercially available standards the method was developed, finally permitting their baseline separation in less than 10 min. The optimum setup showed to be a Viridis HSS C18 SB column, supercritical carbon dioxide and methanol. The compounds could be assigned in the extracts either at 210 nm or by MS, for which no modifications except of an additional sheath liquid (0.1 % acetic acid in methanol) were required. The determined lignan patterns were typical for S. chinensis, with schisandrol A being the most abundant compound, followed by schisandrin B or schisandrol B. As method validation results also complied well with the requirements the here presented method is definitely an interesting alternative to established techniques like UHPLC for the analysis of lignans in Schisandra 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.

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 "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.

[Decipherment of herb medicine Nanwuweizi and Wuweizi in Chinese ancient literature].

Nanwuweizi( Schisandrae Sphenantherae Fructus) and Wuweizi( Schisandrae Chinensis Fructus) have long-term history of use as common traditional Chinese medicines since the Eastern Han Dynasty( AD.25-220 year).However their information are always confused in ancient literature because they were both used as " Wuweizi". Nanwuweizi and Wuweizi are faced with problems such as confused distribution of producing areas,unclear source plants and efficacy characteristics,which limit modern resource development and application. Based on ancient literatures of materia medica,this study conducted a systematic review from several aspects,i.e. the name,distribution of producing areas,source plants,efficacy characteristics and processing of the two medicines in ancient time. This study clarified five main aspects,as following,ancient production areas and corresponding modern distribution areas; source plants used for medicinal purposes in ancient time; application period and application scope; efficacy characteristics in clinical application;processing method. This study provides a reference for evaluating the quality and for their clinical application and reasonable development of Nanwuweizi and Wuweizi.

Detecting Schisandrae Chinensis Fructus and Its Chinese Patent Medicines with a Nucleotide Signature.

Schisandrae Chinensis Fructus (Wuweizi) is often adulterated with Schisandrae Sphenantherae Fructus (Nanwuweizi) in the herbal market. This adulteration is a threat to clinical treatment and safety. In this study, we aimed to develop a nucleotide signature for the identification of Wuweizi and its Chinese patent medicines based on the mini-DNA barcoding technique. We collected 49 samples to obtain internal transcribed spacer 2 (ITS2) sequences and developed a 26-bp nucleotide signature (5'-CGCTTTGCGACGCTCCCCTCCCTCCC-3') on the basis of a single nucleotide polymorphism (SNP) site within the ITS2 region that is unique to Wuweizi. Then, using the nucleotide signature, we investigated 27 batches of commercial crude drug samples labeled as Wuweizi and eight batches of Chinese patent medicines containing Wuweizi. Results showed that eight commercial crude drug samples were adulterants and one of the Chinese patent medicines contained adulterants. The nucleotide signature can serve as an effective tool for identifying Wuweizi and its Chinese patent medicines and can thus be used to ensure clinical drug safety.

Linking resource supplies and price drivers: Lessons from Traditional Chinese Medicine (TCM) price volatility and change, 2002-2017.

ETHNOPHARMACOLOGICAL RELEVANCE: Worldwide, one of the drivers of substitution and adulteration is the cost of the natural resources (plants, animals, fungi) that are ingredients of traditional medicines. Relatively few studies have been done that link prices of traditional medicine ingredients to what drives changes in price, yet this is an important topic. Theoretically, prices have been widely considered as an economic indicator of resource scarcity. Rare, slow growing medicinal plants sell for high prices and common, less popular species for low prices. Price levels also influence the viability of farming vs. wild harvest (and incentives to overharvest high value species when tenure is weak). Prices can also influence the harvesting or buying behaviour of harvesters, traders or manufacturers. When prices are high, then there is a greater incentive to use cheaper substitute species or adulterants. As previous studies on herbal medicine ingredients have shown, adulteration applies in a wide variety of cases, including to some Traditional Chinese Medicine (TCM) species. AIM OF THE STUDY: The aim of this study was to gain a better understanding of which factors influenced changes in the market prices of document prices for four popular, but very different traditional Chinese medicine (TCM) species (2002 - 2017). MATERIALS AND METHODS: Fluctuations in market prices were followed over a 15-year period (2002-2017) for four very different TCM ingredients: two plant species (one wild harvested for fruits (Schisandra sphenanthera Rehder & E.H. Wilson) the other in a transition from wild harvest to cultivation (Paris polyphylla Smith), an animal species (the Tokay gecko (Gekko gecko L.)) and the entomophagous "caterpillar fungus" (Ophiocordyceps sinensis (Berk). G.H. Sung, J.M. Sung, Hywel-Jones & Spatafora). RESULTS: High prices of medicinal plants are widely considered to reflect resource scarcity. Real-time market prices for three of the four very different TCM species we studied all showed major price fluctuations. The exception was P. polyphylla, whose wild populations are widely known to be increasingly scarce, where there was a steady increase in price, with few fluctuations in the upward price trend. The three other species showed significant price fluctuations. These were driven by multiple factors. Ecological and biogeographic factors that influence abundance or scarcity of supply certainly played a role. But other factors were also influential. These included both national and global economic factors (the influence of the Global Financial Crisis (GFC)), national policy changes that in turn influenced businessmen giving expensive gifts (that included O. sinensis)), climate change (influencing fruiting success of S. sphenanthera), price speculation by traders and lack of information (e.g: reduction in G. gecko prices due to traders incorrectly believing that domestication would increase supplies). CONCLUSIONS: Price fluctuations in the four TCM species we examined are influenced by many factors and not just resource scarcity. And the situation is more complex than the trajectory based on Homma's (1992) model, where he predicted that higher prices would result in a shift to cultivation, thus replacing wild harvest. In case of both O. sinensis and P. polyphylla, Homma (1992, 1996) was right in terms of scarcity and high prices stimulating a major investment in cultivation (P. polyphylla) and artificial production (O. sinensis). But in both cases, intensive production through cultivation or artificial propagation do not yet occur on a large enough scale to reduce harvest of wild stocks. Substitution and adulteration occur with all four species. Improving information to medicinal plant traders on the supply status of TCM stocks, whether from wild harvest or from cultivation could benefit product quality, cultivation initiatives and conservation efforts.

Decipherment of herb medicine Nanwuweizi and Wuweizi in Chinese ancient literature / 中国中药杂志

Nanwuweizi( Schisandrae Sphenantherae Fructus) and Wuweizi( Schisandrae Chinensis Fructus) have long-term history of use as common traditional Chinese medicines since the Eastern Han Dynasty( AD.25-220 year).However their information are always confused in ancient literature because they were both used as " Wuweizi". Nanwuweizi and Wuweizi are faced with problems such as confused distribution of producing areas,unclear source plants and efficacy characteristics,which limit modern resource development and application. Based on ancient literatures of materia medica,this study conducted a systematic review from several aspects,i.e. the name,distribution of producing areas,source plants,efficacy characteristics and processing of the two medicines in ancient time. This study clarified five main aspects,as following,ancient production areas and corresponding modern distribution areas; source plants used for medicinal purposes in ancient time; application period and application scope; efficacy characteristics in clinical application;processing method. This study provides a reference for evaluating the quality and for their clinical application and reasonable development of Nanwuweizi and Wuweizi.

Rapid Classification and Identification of Chemical Components of Schisandra Chinensis by UPLC-Q-TOF/MS Combined with Data Post-Processing.

Schisandra chinensis (known in Chinese as WuWeiZi, WWZ) has observable effects such as astringing the lung to stop coughs, arresting sweating, preserving semen and preventing diarrhea. The major components of WWZ include lignans, triterpenoids, organic acids and fatty acids. In this paper, a reliable method for the rapid identification of multiple components in WWZ by their characteristic fragments and neutral losses using UPLC-Q-TOF/MS technology was developed. After review of the literature and some reference experiments, the fragmentation pattern of several compounds were studied and summarized. Then, according to the corresponding characteristic fragments coupled with neutral losses in the positive or negative ion mode produced by different types of substances a rapid identification of target compounds was achieved. Finally, a total of 30 constituents of WWZ were successfully identified, including 15 lignans, nine triterpenoids, three organic acids and three fatty acids. The method established in this study not only provides a comprehensive analysis of the chemical ingredients of WWZ, providing a basis for further phytochemical studies on WWZ but also provides a more efficient way to solve the problem of identification of complex chemical constituents in traditional Chinese medicines.

A Network Pharmacology-Based Study on the Hepatoprotective Effect of Fructus Schisandrae.

Fructus schisandrae (Wuweizi in Chinese), a common traditional Chinese herbal medicine, has been used for centuries to treat chronic liver disease. The therapeutic efficacy of Wuweizi has also been validated in clinical practice. In this study, molecular docking and network analysis were carried out to explore the hepatoprotective mechanism of Wuweizi as an effective therapeutic approach to treat liver disease. Multiple active compounds of Wuweizi were docked with 44 protein targets related with viral hepatitis, fatty liver, liver fibrosis, cirrhosis, and liver cancer. A compound-target network was constructed through network pharmacology analysis, predicting the relationships of active ingredients to the targets. Our results demonstrated that schisantherin, schisandrin B, schisandrol B, kadsurin, Wuweizisu C, Gomisin A, Gomisin G, and angeloylgomisin may target with 21 intracellular proteins associated with liver diseases, especially with fatty liver disease. The CYP2E1, PPARα, and AMPK genes and their related pathway may play a pivotal role in the hepatoprotective effects of Wuweizi. The network pharmacology strategy used provides a forceful tool for searching the action mechanism of traditional herbal medicines and novel bioactive ingredients.

Study on fingerprint of HPLC of crud drug, intermediate, and finished products of Wuweizi Syrup / 中草药

Objective: To establish the HPLC fingerprint and analyze the correlation of Schisandrae Fructus (SF), intermediate, and finished products of Wuweizi Syrup. Methods: The HPLC method was used with the condition that the column was ACE5-C18 (250 mm × 4.6 mm, 5 μm); The mobile phase was eluted with gradient by acetonitrile-water; The flow rate was 1.0 mL/min; The column temperature was 30℃; The detection wavelength was set at 218 nm. Results: The content was determined and the fingerprint was established for SF from Changbai Mountain, intermediate, and finished products of Wuweizi Syrup; Eighteen common peaks of SF and 12 common peaks in intermediate and finished products of Wuweizi Syrup were marked. The chemical composition and content in ten batches of SF from Changbai Mountain were stability; The similarity from the same manufacturer, but different batches of the intermediate and finished product is greater; The correlation between medicinal materials and intermediates as well as finished products is better. Conclusion: The established fingerprints have better reproducibility, which can be used for the quality control of Wuweizi syrup with good precision, accuracy, and reproducibility.

Optimization of phenolic antioxidant extraction from Wuweizi (Schisandra chinensis) pulp using random-centroid optimazation methodology.

The extraction optimization and composition analysis of polyphenols in the fresh pulp of Wuweizi (Schisandra chinensis) have been investigated in this study. The extraction process of polyphenols from Wuweizi pulp was optimized using Random-Centroid Optimization (RCO) methodology. Six factors including liquid and solid ratio, ethanol concentration, pH, temperature, heating time and extraction times, and three extraction targets of polyphenol content, antioxidant activity and extract yield were considered in the RCO program. Three sets of optimum proposed factor values were obtained corresponding to three extraction targets respectively. The set of optimum proposed factor values for polyphenol extraction given was chosen in further experiments as following: liquid and solid ratio (v/w) 8, ethanol 67.3% (v/v), initial pH 1.75, temperature 55 °C for 4 h and extraction repeated for 4 times. The Wuweizi polyphenol extract (WPE) was obtained with a yield of 16.37 mg/g and composition of polyphenols 1.847 mg/g, anthocyanins 0.179 mg/g, sugar 9.573 mg/g and protein 0.327 mg/g. The WPE demonstrated high scavenging activities against DPPH radicals.

Identification of Schisandra chinensis(Turcz.) Baill.(Beiwuwei) and Schisandra sphenanthera Rehd. Et Wils.(Nanwuwei) by Random Amplified Polymorphic DNA / 中药新药与临床药理

Objective: To establish a new method for the identification of Schisandra chinensis(Turcz.) Baill. (Beiwuwei) and Schisandra sphenanthera Rehd. Et Wils. (Nanwuwei). Methods: Random amplified polymorphic DNA method was applied to screen random primers. Results:Screening from 80 primers,only S429, which can be used to identify Schisandra chinensis(Turcz.) Baill.(Beiwuwei) and Schisandra sphenanthera Rehd. Et Wils. (Nanwuwei) accurately and is of good reproducibility. Conclusion: S429 can be used to identify Schisandra chinensis(Turcz.) Baill. (Beiwuwei) and Schisandra sphenanthera Rehd. Et Wils. (Nanwuwei) accurately.

Clinical Observation on Shenqi Wuweizi Tablet for Anxiety before College Entrance Examination:A Report of 113 Cases / 中医杂志

Objective To observe the effect of ShenQi Wuweizi Tablet (Tablet composed of pilose asiabell root, astragalus root, and schisandra fruit) on anxiety before college entrance examination.Methods Totally 227 patients were randomized into a treatment group (113 cases, treated with Shenqi Wuweizi Tablet, 3 tablets each time and 3 time a day), and a control group (114 cases, treated with alprazolam, 0.4 mg each time and 3 times a day). The treatment course was 6 weeks for both groups. Before treatment and at the end of the 1st, 2nd, 4th and 6th week of treatment, the clinical effect was assessed with Hamilton Anxiety Scale (HAMA) and Clinical Global Impression (CGI), and Self-Rating Anxiety Scale (SAS). And at the same time, the untoward reactions was also observed.Results At the end of the 1st week of treatment, the HAMA score was significantly decreased in the control group (P