Analysis of Lavandulyl Flavonoids from Sophora flavescens with Antiinflammatory Activity Based on Molecular Network Technology

[Objectives]This study was conducted to screen lavandulyl flavonoids with anti-inflammatory activity from Sophora flavescens.[Methods]35 compounds were screened from traditional Chinese medicine S.flavescens using the nitric oxide(NO)anti-inflammatory activity model.[Results]Five components,8(xanthohumol),13(kurarinol),27(4-methoxysalicylic acid),28(b-resorcic acid)and 30(b-resorcic acid),exhibited significant anti-inflammatory activity,with IC 50 values of 5.99,4.76,6.96,3.41 and 5.22μM,respectively.Especially,8(xanthohumol)and 13(kurarinol)were typical lavandulyl flavonoids in S.flavescens,which were worth further exploration.Furthermore,UPLC-Q-Exactive and GNPS molecular networking technique were used for rapid analysis of lavandulyl flavonoids from S.flavescens.A total of 15 components were identified.[Conclusions]This work lays a theoretical foundation for further separation and analysis of lavandulyl flavonoids with anti-inflammatory activity from S.flavescens.

Mechanisms of Sophora flavescens in the treatment of cervical squamous cell carcinoma based on comprehensive biological analysis,network pharmacology,and experimental verification

Objective:This study used comprehensive bioinformatics analysis and network pharmacology analysis to investigate the potentially relevant mechanisms of Sophora flavescens against cervical squamous cell carcinoma.Methods:Consistently altered genes involved in cervical squamous cell cancerization were analyzed in the GEO database.The chemical ingredients and target genes of Sophora flavescens were explored using the TCMSP database.We obtained the potential therapeutic targets of Sophora flavescens by intersecting the above genesets and validated them in the GEPIA database.The interaction between Sophora flavescens and target genes was predicted by molecular docking.RT-qPCR was used to verify the changes of target genes in HeLa cells treated with Sophora flavescens.Single-gene GSEA functional analysis were performed to determine the molecular mechanisms.Results:Fifteen genes related to the transformation of cervical squamous cell carcinoma were identified,among which AR and ESR1 were confirmed as targets for kaempferol,wighteone,formononetin,and phaseolinon.These compounds are the active ingredients in Sophora flavescens.Low expressions of AR and ESR1 correlate with a poor prognosis,while Sophora flavescens treatment increases the expression of AR and ESR1 in HeLa.GSEA analysis showed that AR and ESR1 mainly participate in the epithelial-mesenchymal transition in cervical squamous cell carcinoma.Conclusion:Sophora flavescens exert anti-tumor effects by targeting AR and ESR1,which may regulate cancer metastasis.

Two New Antibacterial Flavanones from Sophora flavescens

Two new lavandulylated flavanones,（2R, 3R）-8-1avandulyl-2＇-methoxy-5, 7, 4＇-trihydroxyflavanonol （1） and 8-1avandulyl-5,7, 4＇-trihydroxyflavonol （2）,were isolated from the dry roots of S opharaflavescens.Their structures were eiucidated on the basis of spectroscopic data. Compounds 1 and 2 exhibited significant antibacterial activities.

Research progress on flavonoids from the roots of Sophora flavescens Alt.and their biological activities

Sophora flavescens Alt.,a perennial herb of Sophora in Leguminosae family,was widely distributed in the north and south of China.Its root is utilized as a traditional Chinese medicine to clear heat,drain dampness,kill insect and promote urination by virtue of bitter and cold.Its chemical components are complex and diverse,mainly including alkaloids and flavonoids.Modern pharmacological studies have found that prenylflavonoids and quinolicidine alkaloids are the characteristic chemical constituents and the main active ingredients,with anti-inflammatory,anti-tumor,anti-oxidation,and other various biological activities.This study reviewed the chemical constituents of S.flavescens and the pharmacological effects of flavonoids in it in order to provide reference for further development.

Exploring the anti-diabetic effects and the underlying mechanisms of ethyl acetate extract from Sophora flavescens by integrating network pharmacology and pharmacological evaluation

Background:Sophora flavescens,a traditional Chinese herb medicine,has been used to prevent and cure type 2 diabetes mellitus(T2DM)both in folk medicine and medical institutions.Modern pharmacological studies have also demonstrated that the flavonoids obtained from Sophora flavescens ethyl acetate extract(SFE)exhibited potential anti-diabetic activity.Our previous study elucidated that SFE exerts anti-T2DM effects by regulating the host-microbial metabolic axis.In the present study,we further explored the pharmacodynamic effect and the potential targets of the anti-T2DM activity of SFE by integrating network pharmacology and pharmacological evaluations.Methods:The diabetic rat model was created by streptozotocin and oral administration with SFE for 8 weeks.Then,the T2DM-related index was estimated to assess the interventional effect of SFE.Network pharmacology was applied to identify the likely targets and the pathways modulated by SFE components.Furthermore,Western blotting was applied to verify the prediction.Results:Pharmacological evaluation in vivo revealed that SFE could markedly improve the blood glucose,serum insulin secretion,insulin resistance,liver glycogen synthesis,and the liver tissue structure in T2DM rats.Through network pharmacology analysis,101 active compounds of SFE and 114 targets belonging to 128 pathways were identified.The insulin,TNF,IL-6,and PI3K/Akt pathway may be the key targets and pathway.Based on the results of network pharmacology analysis,IRS/PI3K/Akt and IKK/NF-κB/TNF pathways were selected for further validation.Subsequently,experimental results of Western blotting confirmed that SFE may exert anti-T2DM effects by modulating the IRS/PI3K/Akt and IKK/NF-κB/TNF pathways.Conclusion:SFE may protect the T2DM rats by relieving the insulin resistance and inflammation through regulating the IRS/PI3K/Akt and IKK/NF-κB/TNF pathways.The results of the present study would improve the comprehension of the pharmacological basis of SFE against T2DM and provide a theoretical basis for the clinical use of Sophora flavescens.

Uncovering the mechanism of Sophora flavescens in the treatment of glioma based on network pharmacology

Glioma is a common primary intracranial tumor with high mortality and postoperative recurrence.Developing efficient therapies with lower toxicity is urgently needed.Sophora flavescens(SF)is a common Chinese medicine used to treat eczema,wet ulcers and itchy skin.Modern pharmacological studies have showed that SF has anti-glioma effects,but the mechanism of action remains unclear.This study aims to reveal the pharmacological mechanism of SF in treating glioma.The active components and related targets of SF were obtained from TCMSP.Genecard and Online Mendelian Inheritance in Man(OMIM)databases were used to explore the therapeutic targets for glioma.By making Venn diagram,we obtained 132 common targets of compounds and diseases.STRING databases and Cytoscape were used to construct diagrams of Protein-Protein Interaction(PPI)networks.Through the construction of PPI network,potential targets with degree value greater than the median were taken as core targets for further analysis.A total of 66 core targets were screened out.The degree values of TP53,HSP90AA1,MAPK1,and AKT1 were higher,indicating that these genes played important roles in this network.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were used to further discover the mechanism of active components in SF.Through enrichment analysis,it was found that the core targets were mainly enriched in PI3K/AKT signaling pathway,cell senescence related signaling pathway and IL-17 signaling pathway.In order to further explore the binding between active components and selected target,molecular docking was carried out.Finally,SwissADME was used to determine whether the compound could cross the blood-brain barrier.Based on network pharmacology,we speculated that matrine and formononetin in SF might inhibit PI3K/AKT signaling pathway and cellular senescence related signaling pathway by targeting AKT1,TP53,MAPK1 and other key targets.In summary,this study preliminarily explored the target and mode of action of SF in the treatment of glioma,laying the foundation for further research on its mechanism.

Network-pharmacology and molecular docking-based investigation of mechanism of Sophora flavescens on cancer and inflammation

Objective:In order to explore the systematical regulatory mechanism of Kushen(Sophora flavescens,SF)on inflammation and cancer,we analyzed inter-molecular interactions between herbal ingredients of SF and human inflammation and cancer through network-pharmacology and molecular docking-based approaches.Methods:Firstly,ingredients and potential targets were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,GeneCards database,Therapeutic Targets Database and Online Mendelian Inheritance in Man database.Then,protein-protein interaction network and medicine-ingredient-target-disease network were established and analyzed via STRING and Cytoscape.Surflex-dock was performed by SybylX-2.0.Finally,functional enrichment and pathway enrichment were achieved by Gene Ontology database and Kyoto Encyclopedia of Genes and Genomes database.Results:The results showed that 113 components of SF and 53 potential targets were related in the study.SF exerts anti-inflammatory and anti-cancer mechanism through key targets located in nucleus,such as JUN,MYC,RELA,NCOA,PPARG which may trigger the NF-κB pathway,the Bcl-2/Bax pathway and other pathways to effect DNA transcriptional activity.Conclusions:The study predicted the mechanism of SF on cancer and inflammation.According to the results,we suggest that the ingredients of SF effect on DNA bingding and transcription in nuclear receptors-like JUN,MYC,RELA,NCOA,PPARG.the receptors trigger several pathways including NF-κB pathway,the Bcl-2/Bax pathway and others.Eventually,it regulats inflammatory factors and cell proliferation,senescence and apoptosis.

Comparison on the Origin and Development of Sophora flavescens in China and Japan

Sophora flavescens , which was first recorded in Shen Nong s Herbal Classic , has the functions of clearing away heat, drying dampness, killing parasites and promoting urination. In its long-term application, traditional Chinese medicine has gradually deepened its understanding of the origin and efficacy of S. flavescens . In order to explore changes in the origin and efficacy of traditional Chinese herb S. flavescens in the history of China and Japan, in this paper, the origin and development of S. flavescens in China and Japan were compared from three aspects: the origin, medicinal efficacy, and processing methods of S. flavescens in China and Japan, hoping to provide clear understanding of the medicinal plant S. flavescens .

Synergistic Action between Jasmonic Acid and Nitric Oxide in Inducing Matrine Accumulation of Sophora flavescens Suspension Cells

Secondary metabolites not only play important ecological roles in plants but also are important pharmaceutical and source compounds for derivative synthesis. Production of plant secondary metabolites is believed to be controlled by the endogenous signal network of plants. However, the molecular basis is still largely unknown. Here we show that matrine production of Sophora flavescens Ait. cells treated with low levels of jasmonic acid （JA） and nitric oxide （NO） is significantly increased although treatment with low concentrations of JA or NO alone has no effects on matrine production, showing that JA and NO may act synergistically in triggering matrine production. Moreover, treatment with NO triggers lipoxygenase （LOX） activity and enhances JA levels of the cells, showing that NO may activate the endogenous JA biosynthesis of S. flavescens cells. External application of JA induces nitric oxide synthase-like activities and stimulates NO generation of S. flavescens cells, which suggests that JA may trigger NO generation of the cells. Thus, the results reveal a mutually amplifying reaction between JA and NO in S. flavescens cells. Furthermore, JA and NO inhibitors suppress not only the mutually amplifying reaction between JA and NO but also the synergistic effects of NO and JA on matrine production. Therefore, the data demonstrate that the synergistic action of JA and NO in inducing matrine production might be due to the mutually amplifying reaction between JA and NO in the cells.

Five Novel Pterocarpan Derivatives from Sophora flavescens

Main observation and conclusion Five novel pterocarpan derivatives(1—5)and three known pterocarpans were isolated from the roots of Sophora flavescens Ait.Their structures were elucidated based on extensive spectroscopic data(UV,IR,1D and 2D NMR,and HR-ESI-MS).Notably,compounds 1—5 were the first reported natural products,which possess unique pterocarpan-glucose-long chain aliphatic acid structures.The kinds of aliphatic acids were determined by comparison with authentic aliphatic acids using HPLC-ELSD analysis after acid hydrolysis.The absolute configurations of 1—5 were confirmed by analysis of their ECD spectra.All compounds were evaluated against lipopolysaccharide(LPS)-induced TNF-αproduction in RAW264.7 cells,and compound 1 showed significant inhibitory activity with inhibition rate of 86.22%,IC_(50)=0.79μmol/L(dexamethasone as positive control,inhibition rate of 71.38%).

Simultaneous determination of berberine,matrine and oxymatrine in traditional Chinese medicines by using nonaqueous capillary electrophoresis

A rapid method for the simultaneous determination of berberine(BBR),matrine(MT)and oxymatrine(OMT)by nonaqueous capillary electrophoresis(NACE)was developed.Optimum separation of the analytes was obtained on a 50cm×50μm i.d.fused-silica capillary using a non-aqueous buffer system of 70mM ammonium acetate,7.0% acetic acid and 10% acetonitrile at 25kV and 20℃.The relative standard deviations(R.S.D.)of the migration times and peak areas of the three active components were 0.06%-0.20% and 0.12%-3.41% for berberine,0.11%-0.60% and 0.74%-1.63% for matrine,0.15% and 0.45% for oxymatrine,respectively.Detection limits of berberine,matrine and oxymtrine were 0.18μg/mL,4.08μg/mL and 4.16μg/mL,respectively.In the tested concentration range,good linear relationships(0.9992 for berberine,0.9988 for matrine and 0.9988 for oxymatrine)were observed.The linear calibration ranges were 0.45-360.0μg/mL for berberine,8.16-408.0μg/mL for matrine and 20.8-416.0μg/mL for oxymatrine.This method has been successfully applied to the phytochemical analysis of alkaloids extracts from two commonly used traditional Chinese herbal drugs:Sophora flavescens Ait.(Kushen)and Cortex phellodendri chinensis(Huangbai)and their medicinal preparations.

Simultaneous Determination of Two Types of Active Components with Different Physicochemical Properties in Kushen by HPLC-UV

A simple, reliable and accurate high performance liquid chromatography(HPLC) coupled with a UV detector at varied wavelength was developed for the quantitation of two types of anti-carcinogenic compounds, namely, Kushen alkaloids(KS-As) and Kushen flavonoids(KS-Fs), in Kushen. Their remarkable difference in physicochemical properties was circumvented by integrated optimization of column and mobile phase. The separation was achieved on an alkaline-resisting C18 column via gradient elution with acetonitrile(ACN) and 0.025%(volume ratio) diethylamine(DEA) in water as mobile phase. The flow rate was 0.7 mL/min, and the column temperature was maintained at 35 °C. UV detection wavelength was set at 225 nm to monitor KS-As, and then changed to 335 nm for KS-Fs at 26th min, which was kept for 30 min before returning to the original wavelength. All the calibration curves show good linearity(r20.999) within test ranges. The intra- and inter-day precision values of these components were less than 3.82%, and the average recovery rates obtained were in a range of 97.24%―102.64% for all the samples with RSD below 3.94%. This assay was successfully applied to determining the major active components in commercial herbal samples of Kushen. Moreover, the possible relationship between sample and the geographical region was visualized based on the contents of the 6 compounds in the light of principal component analysis(PCA). The results suggest that the proposed method facilitates the quality control of Kushen.

苦参硬实性及硬实相关基因SfHs1-1的克隆与表达分析

硬实性是种子休眠类型之一,为研究苦参(Sophora flavescens Ait)种子硬实性并探究其分子机制,测定苦参种子的硬实率及发芽率,观察种皮解剖结构,并从苦参中克隆得到SfHs1-1基因(GenBank:MK840-983),进行生物信息学分析。结果表明,苦参具有高硬实率,观察种皮解剖结构发现在发育过程中种皮不断增厚,且栅栏组织所占比例越来越大。克隆获得苦参SfHs1-1基因共1 495 bp,其开放阅读框序列长1 386 bp,编码461个氨基酸;预测Sf Hs1-1蛋白分子量为52 267.15 kD,等电点为9.21,是一个稳定的亲水性蛋白,含有信号肽和多个磷酸化位点,且具有一个保守结构域,属于典型的碱性磷酸酶D家族;系统进化分析表明Sf Hs1-1蛋白与豆科狭叶羽扇豆(Lupinus angustifolius)的PhoD亲缘关系最近。对苦参SfHs1-1基因的表达分析可知,该基因在茎、叶、种皮等各个组织中均有表达,在开花初期表达量基本没有差异,随着种子的生长发育,在第4周种皮中的表达量显著高于其他组织。本研究为苦参硬实性形成机制的探索及改造提供了一定的分子基础。