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Aim To elucidate the molecular mechanism of Sophora tonkinensis Gagnep in improving acute pharyngitis based on network pharmacology, animal experiments and quantitative real-time PCR.Methods The active components and targets of Sophora tonkinensis Gagnep were collected from the database of traditional Chinese medicinal systems databases and analysis platform(TCMSP). Targets related to acute pharyngitis were acquired through GeneCards, OMIM, DrugBank and Disgenet databases. After the common targets of the two were screened, the STRING database was used to construct the protein interaction network, and the Metascape platform was used for pathway analysis. At the same time, Cytoscape software was used to construct a network of "herbal-disease-component-target" and "herbal-disease-component-target-pathway" network. The acute pharyngitis models in rats were established to study the effect of water extract of Sophora tonkinensis Gagnep on acute pharyngitis in rats. Quantitative real-time PCR technology was used to study the effect of Sophora tonkinensis Gagnep on key gene targets in key pathways of pharyngeal tissues in rats with acute pharyngitis. Results In this experiment, 509 related targets of 21 active components of Sophora tonkinensis Gagnep were obtained, 2 167 related targets of acute pharyngitis were obtained, and 194 common targets of Sophora tonkinensis Gagnep and acute pharyngitis were obtained. KEGG pathway analysis screened 344 related signaling pathways, indicating that IL-17 signaling pathway, NF-kappa B signaling pathway and leukocyte transendothelial migration pathway might play a key role in the improvement of acute pharyngitis by Sophorae tonkinensis Gagnep. Animal experiments showed that the low dose group of Sophora tonkinensis Gagnep water extract had better therapeutic effect on acute pharyngitis. The results of quantitative real-time PCR showed that the low-dose group of Sophora tonkinensis Gagnep significantly down-regulated the expression levels of ITGB2, PIK3CA, PIK3CD and PTPN11 genes in leukocyte transendothelial migration pathway(P<0.05). Conclusions The above results show that Sophora tonkinensis Gagnep has the characteristics of multi-component, multi-target and multi-pathway synergy in improving acute pharyngitis, which provides a theoretical basis for further study on the complex mechanism of Sophora tonkinensis Gagnep in improving acute pharyngitis.
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Acute respiratory infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) had caused a global pandemic since 2019, and posed a serious threat to global health security. Traditional Chinese medicine (TCM) has played an indispensable role in the battle against the epidemic. Many components originated from TCMs were found to inhibit the production of SARS-CoV-2 3C-like protease (3CLpro) and papain-like protease (PLpro), which are two promising therapeutic targets to inhibit SARS-CoV-2. This study describes a systematic investigation of the roots and rhizomes of Sophora tonkinensis, which results in the characterization of 12 new flavonoids, including seven prenylated flavanones (1-7), one prenylated flavonol (8), two prenylated chalcones (9-10), one isoflavanone (11), and one isoflavan dimer (12), together with 43 known compounds (13-55). Their structures including the absolute configurations were elucidated by comprehensive analysis of MS, 1D and 2D NMR data, and time-dependent density functional theory electronic circular dichroism (TDDFT ECD) calculations. Compounds 12 and 51 exhibited inhibitory effects against SARS-CoV-2 3CLpro with IC50 values of 34.89 and 19.88 μmol·L-1, repectively while compounds 9, 43 and 47 exhibited inhibitory effects against PLpro with IC50 values of 32.67, 79.38, and 16.74 μmol·L-1, respectively.
Subject(s)
Flavonoids/chemistry , SARS-CoV-2 , Rhizome , COVID-19 , Peptide Hydrolases , Antiviral Agents/chemistryABSTRACT
OBJECTIVE To establish the fingerprints of Ardisia crenata， Sophora tonkinensis and their couplet medicines， and to determine the contents of five components in them. METHODS Using water as solvent， single lyophilized powder of A. crenata and S. tonkinensis and combined lyophilized powder of their couplet medicines were prepared by combining lyophilization technology. The fingerprints of three lyophilized powder samples were established by using high-performance liquid chromatography （HPLC）， and the contents of 5 kinds of components such as gallic acid were determined simultaneously. RESULTS There were 5， 10 and 14 common peaks in the fingerprints for single lyophilized powder of A. crenata and S. tonkinensis and combined lyophilized powder of their couplet medicines； the similarities of them with the control fingerprints were all greater than 0.90. For combined lyophilized powder of couplet medicines， peak 3 Δ 基金项目 国家重点研发计划项目（No.2018YFC1708100）；贵 州省科技计划项目（No.黔科合基础-ZK〔2022〕一般483，No.黔科合成 was identified as gallic acid， peak 4 as matrine， peak 6 as 果〔2021〕一般137）；贵州省教育厅高等学校科学研究项目（青年项目） oxymatrine， peak 8 as bergenin， and peak 14 as trifolirhizin. In single lyophilized powder of A. crenata， the average contents of gallic acid and bergenin were 0.499 3 and 4.962 6 mg/g， respectively. In single lyophilized powder of S.tonkinensis， the average contents of matrine， oxymatrine and trifolirhizin were 3.046 0， 2.336 6 and 0.278 6 mg/g， respectively. In combined lyophilized powder of couplet medicines， the average contents of gallic acid， matrine， oxymatrine， bergenin and trifolirhizin were 0.560 6， 2.548 7， 1.382 2， 5.960 7 and 0.279 1 mg/g， respectively. The transfer rates were 8.87%-513.19%. CONCLUSIONS The established fingerprint and content determination methods are stable and feasible， and can be used for the quality control of A. crenata and S. tonkinensis and their couplet medicines. The average contents of matrine and oxymatrine in combined lyophilized powder of A. crenata-S. tonkinensis couplet medicines are decreased.
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Aromatic constituents from rhizomes of Sophora tonkinensis were purified by extensive chromatographic techniques including column chromatography over macroporous resin,MCI,silica gel,weak acid cation exchange resin,Sephadex LH-20,ODS,and semi-preparative HPLC. Twelve aromatic compounds were isolated and identified from the water aqueous extract of the rhizomes of S.tonkinensis. Their structures were elucidated as 4-( 3-hydroxypropyl) phenol( 1),( ±)-4-( 2-hydroxypropyl) phenol( 2),benzamide( 3),( ±)-3-( p-methoxyphenyl)-1,2-propanediol( 4),4-methoxybenzamide( 5),3-hydroxy-1-( 4-hydroxy-3-methoxyphenyl) propan-1-one( 6),tyrosol( 7),( ±)-2,3-dihydroxypropyl benzoate( 8),vanillin alcohol( 9),7,3'-dihydroxy-8,4'-dimethoxyisoflavone( 10),7,4'-dihydroxy-3'-methoxyisoflavone( 11),and 7,3'-dihydroxy-5'-methoxyisoflavone( 12). Compounds 1-9 were firstly isolated from the Sophora genus. Compounds 4,5,10 and 11 can remarkably protect Hep G2 cell against APAP-induced damage at the concentration of 10 μmol·L-1. Compounds 1-12 exhibited no significant activities on the assays of inhibition of LPS-induced NO production in RAW cell lines and NF-κB inhibition.
Subject(s)
Humans , Chromatography, High Pressure Liquid , Hep G2 Cells , Rhizome/chemistry , Sophora/chemistryABSTRACT
OBJECTIVE： To investigate the potential mechanism of Sophora tonkinensis in the treatment of leukemia. METHODS： The active components and their target proteins of S. tonkinensis were searched by the analysis of traditional Chinese medicine system pharmacology platform， and UniProt database and PubMed database were used to query corresponding gene names of target proteins of active components. Cytoscape 3.6.0 software was used to construct compound-target network. The genes related to leukemia were searched by DisGeNET databases， and OmicShare platform was used to screen the intersection genes of the active component targets of S. tonkinensis and leukemia disease targets. STRING database and Cytoscape 3.6.0 software were used to construct PPI network， and topological analysis was performed. GO analysis and KEGG pathway enrichment analysis were performed by using DAVID bioinformatics database. RESULTS： There were 13 active components and 204 target proteins in S. tonkinensis. The components and targets with high node degree included quercetin， kaempferol， PTGS2， PRSS1， CAMKK2， etc. There were 24 intersection genes between the active component target and leukemia target， including IRF1， BCL2， CYP1A1， PIM1， etc. PPI network of the above intersection genes contained 24 nodes and 142 edges， with an average node degree of 6.5 and an average medium of 0.045. The results of GO analysis showed that the biological process of the above-mentioned genes involved in apoptosis signaling pathway in vitro without ligands， negative regulation of apoptosis process， positive regulation of B cell proliferation， etc. Molecule function mainly included that protein homology activity and binding of the same protein. Cell components mainly included extracellular region， mitochondria and so on. KEGG pathway enrichment showed that above-mentioned genes were mainly associated with T cell receptor signaling pathway， JAK/STAT signaling pathway， HTLV-Ⅰ infection. CONCLUSIONS： Through JAK/STAT signaling pathway and HTLV-Ⅰ infection pathway， the active components of S. tonkinensis may act on PTGS2， PRSS1， CAMKK2 and other targets， and then play a therapeutic role on leukemia， showing the characteristics of multi-component， multi-target and multi-channel.
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Sophorae Flavescentis Radix (Sophora flavescens Ait., SFR) and Sophorae Tonkinensis Radix et Rhizoma (S. tonkinensis Gapnep., STR) are two commonly used traditional Chinese medicines from Sophora (Leguminosae) plants, which are believed to possess similar bioactive components with entirely different clinical applications. In order to find out the characteristic chemical constituents potentially leading to the unique medicinal properties claimed for each of the two closely related TCMs, an HPLC fingerprint method was developed for analyses of the alkaloid and flavonoid constituents of SFR and STR, respectively, which were further evaluated and compared through similarity calculation and hierarchical clustering analysis (HCA). The results from the present study showed that the alkaloid fingerprints of the two herbs were similar, with many components co-existing in both drugs and various batches of samples from different species being mixed together in the HCA dendrogram. However, their flavonoid constituents were totally different with specific fingerprints being yielded for each herb, and further HCA analysis showed that the tested samples could almost be clearly divided into two groups based on their origins of species. The results from the present study indicated that the flavonoid constituents could serve as the differentially diagnostic constituents of SFR and STR and might potentially attributed to their distinct therapeutic effects.
Subject(s)
Alkaloids , Chromatography, High Pressure Liquid , Methods , Discriminant Analysis , Drugs, Chinese Herbal , Flavonoids , Rhizome , Chemistry , Sophora , Chemistry , ClassificationABSTRACT
Sophorae Flavescentis Radix (Sophora flavescens Ait., SFR) and Sophorae Tonkinensis Radix et Rhizoma (S. tonkinensis Gapnep., STR) are two commonly used traditional Chinese medicines from Sophora (Leguminosae) plants, which are believed to possess similar bioactive components with entirely different clinical applications. In order to find out the characteristic chemical constituents potentially leading to the unique medicinal properties claimed for each of the two closely related TCMs, an HPLC fingerprint method was developed for analyses of the alkaloid and flavonoid constituents of SFR and STR, respectively, which were further evaluated and compared through similarity calculation and hierarchical clustering analysis (HCA). The results from the present study showed that the alkaloid fingerprints of the two herbs were similar, with many components co-existing in both drugs and various batches of samples from different species being mixed together in the HCA dendrogram. However, their flavonoid constituents were totally different with specific fingerprints being yielded for each herb, and further HCA analysis showed that the tested samples could almost be clearly divided into two groups based on their origins of species. The results from the present study indicated that the flavonoid constituents could serve as the differentially diagnostic constituents of SFR and STR and might potentially attributed to their distinct therapeutic effects.
Subject(s)
Alkaloids , Chromatography, High Pressure Liquid , Methods , Discriminant Analysis , Drugs, Chinese Herbal , Flavonoids , Rhizome , Chemistry , Sophora , Chemistry , ClassificationABSTRACT
The aim is to systemically review and evaluate the safety of Sophora tonkinensis from the literature on the herbal origin, toxicity record in modern literature and toxicological studies and publications in recent years. By systematic review and analysis, the results showed that its toxicity mainly involved the nervous system, the digestive system and the respiratory system, and respiratory failure may be the direct cause of death. The main symptoms included headache, dizziness, vomiting, nausea, abdominal pain, limbs weakness, palpitation, and chest distress; as well as pale complexion, limbs trembling, convulsions, chills, high heart rate, fall of blood pressure, shock, and respiratory failure to death in severe cases. High dose and long term medication may cause serious brain damage, especially in adolescents and children. The authors have proposed to use rationally under guidance of physician and strictly according to the dosage recommended by pharmacopoeia. The patients shall not be credulous about the folk prescriptions and test recipes to use it for,prevention of colds and treatment of sore throat at will. In addition, the researches on the conventional treatment methods for S. tonkinensis poisoning, the toxic substance basis, and toxicity mechanism shall be strengthened in further studies. These efforts will play important role in exerting the drug effect and avoiding side effect.
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Thirteen compounds were isolated from the 95% aqueous EtOH extract of the rhizomes of Sophora tonkinensis by a combination of various chromatographic techniques including column chromatography over silica gel, Sphadex LH-20, MCI, ODS, and semi-preparative HPLC.Their structures were elucidated as 1-(6,7-dihydro-5H-pyrrolo[1,2-a]imidazol-3-yl)ethanone(1), cyclo(Pro-Pro)(2), nicotinic acid(3), p-hydroxybenzonic acid(4), p-methoxybenzonic acid(5), 4-hydroxymethyl-2,6-dimethoxyphenol-1-O-β-D-glucopyranoside(6), coniferin(7), syringin(8),(-)-secoisolariciresinol-4-O-β-D-glucopyranoside(9),(-)-syringaresinol-4-O-β-D-glucopyranoside(10),(-)-syringaresinol-4,4'-di-O-β-D-glucopyranoside(11),(-)-pinoresinol-4,4'-di-O-β-D-glucopyranoside(12), and(6S,9R)-roseoside(13) by their physicochemical properties and spectroscopic data.Compound 1 was a new naturalproduct, and compounds 2,5,6,9,10,12 and 13 were obtained from the Sophora genus for the first time.Compound 1 possessed moderate cytotoxic activity against A549 human tumor cell [IC₅₀（23.05 ± 0.46）μmol•L⁻¹].
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Twelve quinolizidine alkaloids were isolated from Sophora tonkinensis by means of silica gel, preparative MPLC, and preparative HPLC. On analysis of NMR spectroscopic data, their structures were established as 3-(4-hydroxyphenyl)-4-(3-methoxy-4-hydroxyphenyl)-3,4-dehydroquinolizidine(1), lanatine A(2), cermizines C(3), senepodines G(4), senepodines H(5), jussiaeiines A(6), jussiaeiines B(7),(+)-5α-hydroxyoxysophocarpine(8),(-)-12β-hydroxyoxysophocarpine(9),(-)-clathrotropine(10),(-)-cytisine(11), and (-)-N-methylcytisine(12), respectively. Compounds 1-7 were first isolated from Sophora L. plant. In the in vitro assays,the isolated compounds 1, 3, 6-10 exhibited potent activity against CVB3 with IC₅₀ of 6.40, 3.25, 4.66, 3.21, 0.12, 0.23 and 1.60, and with selective index values(SI=TC₅₀/IC₅₀)of 12.0, 5.6, 13.0, 15.1, 50.1, 26.2, and 23.6, respectively. Compounds 1, 3, and 7 exhibited activity against staphylococcus aureus(ATCC 29213)with MICvalues of 8.0, 3.5, 6.0 g•L⁻¹, respectively. Compounds 1, 3, 7, and 12 exhibited activity against staphylococcus aureus(ATCC 33591)with MIC values of 18.0, 7.5, 8.0, 12.0 g•L⁻¹, respectively. Compounds 2, 6, 7 exhibited activity against Escherichia coli(ATCC 25922) with MIC values of 1.0, 3.2, 0.8 g•L⁻¹.
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AIM@#To develop analytical methods for the identification and determination of the flavonoids in Sophora tonkinensis for comprehensive quality evaluation.@*METHODS@#An HPLC-DAD-ESI-MS method was used for the separation and characterization of the flavonoids in S. tonkinensis, and a liquid chromatographic method was employed to simultaneously determine five major active flavonoids in this crude drug.@*RESULTS@#Seventeen flavonoids were identified, among which, seven were unambiguously identified as trifolirhizin, quercetin, formononetin, macckiain, kurarinone, sophoranone, and sophoranochromene by comparing their retention times, and UV and MS spectra with those of the authentic compounds, and the other ten flavonoids were tentatively identified by comparing their UV and MS/MS spectra with those of literature data. Furthermore, five major active flavonoids, including trifolirhizin, quercetin, maackiain, sophoranone, and sophoranochromene were determined in S. tonkinensis. All calibration curves expressed good linearity (r > 0.999 8) within the test ranges, and the recovery from this method was 96.40%-104.43%. The developed HPLC method was successfully applied to quantitatively determine the five flavonoids in seventeen samples of S. tonkinensis.@*CONCLUSION@#The developed method rapidly characterized the bioactive flavonoids of S. tonkinensis, and could be readily utilized to enhance the quality assurance approaches for this traditional Chinese medicine.
Subject(s)
Chromatography, High Pressure Liquid , Methods , Flavonoids , Chemistry , Molecular Structure , Plant Extracts , Chemistry , Sophora , Chemistry , Spectrometry, Mass, Electrospray Ionization , MethodsABSTRACT
OBJECTIVE: To study the optimal extraction technology of matrine and oxymatrine from Radix Sophorae Tonkinensis. METHODS: The total content of matrine and oxymatrine as the inspection index was determined using high performance liquid chromatography (HPLC). The extraction efficiency of warm immersion extraction, ultrasonic extraction, reflux extraction and accelerated solvent extraction (ASE) was compared. RESULTS: Accelerated solvent extraction was superior to the other three methods. The extraction conditions of accelerated solvent extraction were optimized through single-factor test and orthogonal experimental design. The sequence of influential significance was as follows: extraction times > ethanol concentration > extraction temperature > extraction time. The optimal technology of accelerated solvent extraction was that the ethanol concentration was 40%, extraction temperature was 90°C, the time of extraction was 13 min and the times of extraction was twice. The total content of matrine and oxymatrine from Radix sophorae tonkinensis was (15.5277 ± 0.4316) mg · g-1. CONCLUSION: The experiment has optimized the extraction method and parameters of alkaloid from Radix sophorae tonkinensis, which provides a new method for the extraction of alkaloid from Radix sophorae tonkinensis. Copyright 2012 by the Chinese Pharmaceutical Association.