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Aims@#The current study was aimed to evaluate the antibacterial activity of biogenic synthesized golden nanoparticles from Sophora flavescens Aiton roots aqueous extract against multidrug-resistant (MDR) clinical bacterial isolates.@*Methodology and results@#The green synthesis of gold nanoparticles (AuNPs) was accomplished using S. flavescens roots aqueous extract and examined using many accepted techniques. The antibacterial activity of S. flavescens extract and the aqueous AuNPs at concentrations (7% and 9%) ppm were investigated against two clinical MDR bacteria, including Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Pseudomonas aeruginosa). The findings demonstrate inhibitory activity against the selected MDR bacterial isolates for the aqueous extract of S. flavescens and the aqueous AuNPs noted by the significant decrease in the number of bacteria after treatment with highly significant differences (P≤0.01) compared to the untreated control.@*Conclusion, significance and impact of study@#Sophora flavescens root extracts and their biosynthesized AuNPs with antibacterial activity may find broad applications in fighting MDR pathogenic bacteria and therapeutic manufacturing.
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Anti-Bacterial Agents , Sophora flavescensABSTRACT
OBJECTIVE To establish the fingerprint of Sophora flavescens， and to screen differential components and determine their contents. METHODS HPLC fingerprints of 12 batches of S. flavescens were established by using Similarity Evaluation System of Chromatographic Fingerprints of TCM （2012 edition）； common peaks were identified and their similarities were evaluated. Chemical pattern recognition analysis [cluster analysis （CA），principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）] were performed with SIMCA 14.1 and SPSS 23.0 software， and differential components which influenced the quality of S. flavescens were screen with variable importance in the projection（VIP）＞1 as standard. Meanwhile， the contents of 4 kinds of differential components were determined by the same HPLC method. RESULTS There were 17 common peaks in the fingerprints of 12 batches of S. flavescens，and their similarities were all higher than 0.96. A total of 6 common peaks were identified， i.e. oxymatrine （peak 1）， oxysophocarpine （peak 2）， matrine （peak 10）， trifolirhizin （peak 14）， kurarinone （peak 16） and norkurarinone （peak 17）. Results of CA， PCA and OPLS-DA showed that 12 batches of S. flavescens were divided into 3 categories according to producing area， i.e. S1-S7 （Shangzhou District of Shaanxi Province） were grouped into one category， S8-S10 （Yichuan County of Henan Province） into one category and S11-S12 （Chifeng City of Inner Mongolia） into one category. VIPs of matrine， norkurarinone， kurarinone and oxysophocarpine and the chemical components represented by peak 11 and 9 were all greater than 1. The contents of matrine， norkurarinone， kurarinone and oxysophocarpine in 12 batches of S. flavescens were 2.65-4.93， 1.54-3.44， 9.63-12.94 and 5.08-6.10 mg/g， respectively. CONCLUSIONS HPLC fingerprint of S. flavescens is established successfully in the study， and can be used to screen 6 differential components by combining with chemical pattern recognition analysis， which can provide reference for quality control of S. flavescens.
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italic>Sophora flavescens is a traditional Chinese medicine rich in flavonoids and has wide application potential in drug development and clinical practice. In this study, a total of 227 flavonoids were detected among five tissues of S. flavescens during anthesis using widely targeted metabolomics techniques. There were 137 flavonoids shared by five S. flavescens tissues and 18 root-specific flavonoids. There were 156, 155, 156 and 150 differentially accumulated metabolites identified in stem, leaf, flower, and young pod, respectively, compared with root. Forty-seven potentially active flavonoid components in S. flavescens were identified using the PubChem and SwissADME databases. The 58 potential target proteins for these potentially active components were predicted to be important in the treatment of type 2 diabetes mellitus (T2DM) based on the SwissTargetPrediction and GeneCards database. These 58 target proteins were used to construct a protein-protein interaction network through the STRING database, from which we performed GO and KEGG functional enrichment analysis. The mechanisms by which S. flavescens flavonoids may be useful in the treatment of T2DM was further explored in a multi-level and systematic way based on a "component-target-pathway" network. Finally, ten key potentially effective components were identified and found to be mainly distributed in the roots, flowers, and pods, and their content varied significantly between tissues. The results predict that the key targets of S. flavescens flavonoids in the treatment of T2DM are AKT1, ESR1, EGFR, PIK3R1, TNF and PTGS2, and that they play a hypoglycemic role through the regulation of endocrine resistance, AGE-RAGE, the PI3K-Akt signaling pathway, EGFR tyrosine kinase inhibitor resistance and other signaling pathways. This analysis of the tissue distribution and network pharmacology of S. flavescens flavonoids provides a theoretical basis for further studies on S. flavescens metabolites, the rational development and utilization of the S. flavescens aboveground parts, and initiates a comprehensive exploration of the mechanisms by which S. flavescens can be used in the treatment of T2DM.
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Objective To analyze the main active components and potential molecular mechanism of Sophora flavescens against breast cancer based on network pharmacology and molecular docking. Methods The chemical constituents were collected and screened by TCMSP, ETCM database and literature review. The targets of active ingredients were predicted by Swiss Target Prediction database. Breast cancer-related targets were collected by GeneCards, TTD, Drugbank and OMIM. The anti-breast cancer targets of Sophora flavescens were screened by Venny 2.1.0 software. Cytoscape software was used to construct the network diagram of Sophora flavescens-key active ingredients-targets. STRING database was used to analyze the common targets, and PPI network diagram was constructed. GO function enrichment analysis and KEGG pathway enrichment analysis of key target proteins were performed by DAVID database and Hiplot online platform. Schrodinger software was used to calculate the molecular docking between the active ingredients and targets. Molecular biological methods were used to verify the key targets. Results A total of 36 active components with clear structures were screened from Sophora flavescens. 70 anti-breast cancer targets of Sophora flavescens were screened out. 12 core targets including EGFR, AKT1, ESR1, SRC, CYP19A1, AR and ABCB1 participate in endocrine resistance, EGFR tyrosine kinase inhibitors and estrogen signaling pathways in breast cancer. Moreover, the docking score between the core component and the key target AR is the highest. In vitro experiments showed that the extract of Sophora flavescens can inhibit the proliferation of breast cancer cells, induce cell apoptosis and up-regulate AR protein expression. Conclusion It was revealed that Sophora flavescens plays an anti-breast cancer role by regulating complex biological processes through multiple components acting on multiple targets and signaling pathways. The upregulation of AR protein by Sophora flavescens may become a new therapeutic strategy for the treatment of breast cancer.
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ObjectiveTo explore the antidepressant effect of Sophora flavescens seed extract and its molecular mechanism. MethodA mouse depression model was established by intraperitoneal injection of lipopolysaccharide（LPS）， and normal group， model group， fluoxetine group（2.5 mg·kg-1）， and S. flavescens seed low， medium and high dose groups（200， 400， 800 mg·kg-1） were set up for 7 d of consecutive gavage. Then the antidepressant effect of S. flavescens seed extract was evaluated by using open field test， elevated plus maze test and forced swimming test. Pathological morphological changes in the hippocampal tissue was observed by hematoxylin-eosin（HE） staining. Protein expression levels of G1/S-specific cyclin D1（Cyclin D1）， Wnt1， β-catenin and phosphorylated glycogen synthase kinase-3β（p-GSK-3β） in mouse brain tissues were detected by Western blot. Hippocampal cell apoptosis was detected by terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate（dUTP） nick end labeling（TUNEL）. ResultThe results of mouse behavioral experiments showed that compared with the normal group， the speed of movement in the open field and the distance of movement in the central area of the open field， and the time spent on the open arms of the elevated plus maze were significantly reduced in the model group（P<0.01）， while immobility time in the forced swimming test was significantly increased（P<0.05）. Compared with the model group， the S. flavescens seed medium and high dose groups had increased speed of movement in the open field test and time spent on the open arms of the elevated plus maze test（P<0.05， P<0.01）， and decreased immobility time in the forced swimming test（P<0.05）， the distance of movement in the central area of the open field test increased in the high dose group（P<0.05）. HE staining results showed that compared with the normal group， the hippocampal neuron structure of mice in the model group was damaged. Compared with the model group， after treatment of S. flavescens seed extract， the pathological state of the mouse hippocampal neuron structure was alleviated， and the neurons increased， were neatly arranged， and the cytoplasm was clear. Western blot results showed that the protein expression levels of Wnt1 and β-catenin in mouse brain tissue were significantly decreased（P<0.01）， while the protein expression levels of Cyclin D1 and p-GSK-3β were significantly increased（P<0.01） after LPS injection. Compared with the model group， protein expression levels of Wnt1 and β-catenin in brain tissue of S. flavescens seed medium and high dose groups were significantly increased（P<0.01）， while the protein expression levels of Cyclin D1 and p-GSK-3β were significantly decreased（P<0.01）. TUNEL staining results showed that the hippocampal cell apoptosis rate in the model group was significantly increased compared with that of the normal group（P<0.01）， while the hippocampal cell apoptosis rate in the S. flavescens seed medium and high dose groups was significantly decreased compared with that of the model group（P<0.01）. ConclusionS. flavescens seed extract can effectively improve the severity of depression in LPS-induced depressed mice， and its molecular mechanism is related to the regulation of neuroinflammation and hippocampal neuronal apoptosis mediated by Wnt/β-catenin signaling pathway.
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Objective To establish the quality standard of compound Yuhong suppository. Methods Angelica dahurica, colophony and Sophora flavescens Alt. were identified by thin layer chromatography（TLC）method. The contents of sulfadiazine and dyclonine hydrochloride were determined by HPLC with diode array detection method. The mobile phase was methanol-0.02 mol/L potassium dihydrogen phosphate (adjusted to pH 3.3 with phosphoric acid) for gradient elution. The detection wavelength was 280 nm for sulfadiazine and dyclonine hydrochloride. Results The three Chinese traditional medicines were identified by TLC with clear spots. The linear ranges of sulfadiazine and dyclonine hydrochloride were good in 12.40-99.20 μg/ml (r=0.999 9) and 2.56-20.48 μg/ml (r=0.999 9). The average recovery was (99.21±0.43) % (n=9) and (99.54±0.68) % (n=9). Conclusion This method is accurate, sensitive, and reproducible. It can be used as a standard method for the quality control of compound Yuhong suppository.
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Introduction @#The roots of Sophora Flavescentis is one of the key ingredient in Norbu 7 traditional medicine, the bioactive compound being quinolizidine alkaloids, matrine and oxymatrine. A high performance liquid chromatography (HPLC) method was used to determine matrine, oxymatrine simultaneously in the traditional medicine. The HPLC method was tested and validated for selective determination of matrine and oxymatrine in the Norbu 7 granule. The proposed method was validated for linearity, precision (system precision, method precision, intermediate or inter- day precision) and accuracy, stability in analytical solution, system suitability and ruggedness.@*Goal@#The goal of this study was to develop validated determination method of alkaloid in Norbu 7 granule for quality control.@*Material and Method@#HPLC analysis was performed on Chromecore amino bonded silica gel as the stationary phase (250 mm : 4.6 mm i.d., 5µm) using mixture of acetonitrile, dehydrated ethanol and 3% phosphoric acid (80:10:10) as the mobile phase, 220 nm as the UV light detection. </br> The research methodology was approved by Research Ethic Review Committee of Mongolian University of Pharmaceutical Science on 16th of November, 2020. @*Results@#The calibration curve of oxymatrine showed good linearity (R2=0.9955) within the established range of 8 – 64 µg/ml. The limit of detection (LOD) and quantification (LOQ) were 10.13 µg/ml and 30.71 µg/ ml respectively. Good results were achieved with repeatability (%RSD < 2.0) and recovery (93.08 – 104.32%).@*Conclusion@#The method was found to be selective, accurate, reproducible and the other components did not interfere with determinations. It was successfully used to analyze the granule traditional medicine with 7 different plant formulation and additives. The HPLC method can be used to evaluate and control quality, stability of Norbu 7 granules.
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Sophorae Flavescentis Radix,derived from the root of Sophora flavescens in the Leguminosae family,has been widely used in the medicine,agriculture,animal husbandry,and daily chemical industry. A pharmacophore model-based method for rapid discovery of tyrosinase inhibitors( TIs) from S. flavescens was established by molecular docking under Lipinski rules,and verified by enzyme assays. Briefly,the chemical constituent database of S. flavescens( CDSF) was established based on the previous papers. Theoptimal pharmacophore model( OPM) was constructed by DS 2019 on the basis of known active TIs. Eighty-three hits predominated by flavonoids having higher fitting scores with OPM than the positive control were screened out,and subjected to molecular docking based on the three-dimensional structure of tyrosinase crystal protein. The potential TIs such as kurarinone and nor-kurarinone were rapidly discovered from the compounds with higher docking scores than the positive control under the Lipinski rules. The results were verified by the in vitro enzyme assays. The inhibition activities of tyrosinase from non-medicinal parts of S. flavescens were also tested to explore the relationship between the inhibition activity and chemical compositions. This study is expected to provide data support for the comprehensive application and development of S. flavescens and also a new method for the rapid discovery of active substances or functional constituents in the complex systems.
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Animals , Flavonoids , Molecular Docking Simulation , Monophenol Monooxygenase , Plant Extracts/pharmacology , Plant Roots , SophoraABSTRACT
Objective: To study the molecular mechanisms of antioxidant effect and anti-inflammatory of water extract of Sophora flavescens (WSF) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Methods: The optimum concentration of WSF was evaluated by CCK-8 assay. The inflammatory model was established with LPS by stimulating RAW264.7 cells in vitro. Then all cells were divided into control group, model group, WSF group and WSF control group. The levels of ROS and NO were analyzed with flow cytometry. Subsequently, the expression of iNOS, COX-2, Nrf2, and HO-1 was detected with qRT-PCR and Western blotting. Finally, the pro-inflammatory cytokines IL-6, TNF-α and anti-inflammatory cytokine IL-10 were detected by ELISA. Results: The CCK-8 assay revealed that 0.01 mg/mL WSF did not affect the cell viability. Compared with control group, the LPS-induced inflammatory response could significantly increase the production of NO and ROS, and the IL-6 and TNF-α were also significantly increased (P < 0.05, 0.01, and 0.001). Furthermore, the expression of iNOS and COX-2 were significantly increased (P < 0.01, 0.001), but the expression of Nrf2 and HO-1 were inhibited (P < 0.05). However, compared with model group, the WSF group not only significantly decreased the levels of NO, ROS, IL-6, and TNF-α, but also decreased the expression of iNOS and COX-2 (P < 0.05, 0.01, and 0.001). In contrast, the the level of IL-10 and the expression of Nrf2 and HO-1 were significantly increased (P < 0.05, 0.01, and 0.001). Conclusion: These results suggested that SF exerted protective effect against LPS-induced inflammatory and oxidative responses in RAW 264.7 cells by the activation of the Nrf2/HO-1 pathway.
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The coronavirus disease 2019 (COVID-19), originated in Wuhan city, Hubei Province, China in December 2019, and then quickly spread to most provinces and regions in China and even spread to many countries abroad. COVID-19 is characterized by wide epidemic, strong infectivity, rapid onset and critical condition. In the face of this epidemic, all parts of the country quickly set off a peak in the fight against COVID-19, but no effective drug for COVID-19 has been developed in the short term. Recently, many hospitals have combined traditional Chinese medicine with western medicine in treatment, and the clinical effect is remarkable, which proves the antiviral effect of traditional Chinese medicine. A large number of pharmacological and clinical studies have proved that the Chinese materia medica S. flavescens has significant antiviral effect. In this paper, the mechanism of anti-coronavirus effect of S. flavescens is expounded from multiple pathways, such as type I interferon, NF-κB signal pathway, ERK signal pathway, PI3K/Akt signal pathway and matrine alkaloids, etc. It is intended to provide reference for clinical treatment of coronavirus infection pneumonia and research and development of related drugs of S. flavescens.
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Sophora flavescens is a traditional herb medicine in China. Matrine and oxymatrine are the primary components in S. flavescens with an outstanding anti-cancer potentials. This review aims to summarize the mechanism of tumor-inhibition of matrine and oxymatrine by screening and analyzing the recent literatures. It was found that the molecular mechanisms of tumor-inhibition of matrine and oxymatrine include the regulation of cell cycle progression, induction of cell apoptosis, anti-metastatic and anti-invasion activities, induction of autophagy of tumor cells, reversion of the multidrug resistance in cancer cells, and regulation of metabolic level in cancer cells. Further research on the molecular mechanisms of matrine and oxymatrine could reveal their cellular signaling network and gene regulation mechanism in the anticancer behaviors, so as to find more accurate tumor therapeutic targets, which have important implications not only for new drugs development in clinical application of cancers, but also for the modernizations and internationalization of Chinese medicine.
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Objective: To establish a UPLC-MS/MS method for simultaneous determination of sophoraflavanone G (SFG), kurarinone (Kur), and isoxanthohumol (Iso) in rat plasma using rutin as the internal standard (IS), and to study the pharmacokinetic parameters of total flavonoids (TF) from Sophora flavescens and TF self-microemulsion drug delivery system (TF-SMEDDS) in rats. Methods: Analysis was carried out on an ACQUITY UPLC® HSS T3 C18 column using acetonitrile-0.1% formic acid in water as the mobile phase at a flow rate of 0.2 mL/min. The plasma samples were prepared by liquid-liquid extraction with ethyl acetate. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring with an electro-spray ionization source in negative ionization mode. All statistical analysis was performed using DAS 2.0 software package. Results: The calibration curves of Kur, SFG, and Iso exhibited good linearity (r2 > 0.995 9) over the range of (0.02-1 600.00), (0.015-1 200.000), and (0.01-800.00) ng/mL, respectively. Precision, accuracy, average extraction recovery, and matrix effects were all in line with biological sample analysis requirements. The pharmacokinetic parameters of Kur, SFG, and Iso from TF were as follows: t1/2z (7.04 ±2.46), (4.54 ± 2.13), (4.73 ± 1.67) h, and AUC0-∞ (3 469.57 ± 555.37), (2 524.48 ± 425.83), (1 006.47 ± 85.46) ng∙h/mL. The pharmacokinetic parameters of Kur, SFG, and Iso from TF-SMEDDS were as follows: t1/2z (13.10 ± 2.67), (11.47 ± 4.17), and (12.67 ± 4.97) h, and AUC0-∞ (13 002.49 ± 2 498.09), (8 070.80 ± 2 264.62), (3 918.85 ± 429.76) ng∙h/mL. The relative bioavailabilities of Kur, SFG, and Iso in TF-SMEDDS were approximately 374.76%, 319.70%, and 389.37%, respectively. Conclusion: The UPLC-MS/MS method can be used to study the pharmacokinetics of three components in S. flavescens in rats. The bioavailability of TF-SMEDDS in rats was significantly increased.
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Objective: To detect the effect of alkaloids of Sophora flavescens on biofilm formation in Staphylococcus epidermidis in vitro, and to observe whether total alkaloids of S. flavescens can affect the tolerance of Staphylococcus epidermidis to lactic acid and hydrogen peroxide, then explore the possible anti-microbial mechanism of alkaloids in Sophora flavescens. Methods: The biofilm of Staphylococcus epidermidis was prepared in vitro. We evaluated the effect of alkaloids in S. flavescens on the biofilm-forming ability of Staphylococcus epidermidis via 96-well cell culture microtiter plates with crystal violet staining as well as Congo red experiment. The biofilm formation of Staphylococcus aureus was observed using scanning electron microscope. Then, we measured the change of tolerance to oxidative stress and sensibility to antibiotics for Staphylococcus epidermidis being treated by alkaloids in S. flavescens Ait. qRT-PCR analysis was performed to show the relative transcript level of serp2195 and gpxA-2 upon Staphylococcus epidermidis strain exposed to alkaloids in S. flavescens at different concentrations. Results: Alkaloids in S. flavescens significantly prevented biofilm formation in Staphylococcus epidermidis (P < 0.001) at the concentration of 5.0 mg/mL during crystal violet staining and Congo red experiment, and thus weakened tolerance to oxidative stress and enhanced sensibility to lactate for Staphylococcus epidermidis. Alkaloids in S. flavescens could downregulate the transcript level of serp2195 and gpxA-2. Conclusion: Alkaloids in S. flavescens has a significant inhibitory effect on biofilm formation of Staphylococcus epidermidis, which weakens the tolerance of bacteria to oxidative stress. The mechanism may be realized by downregulating the transcript level of serp2195 and gpxA-2, and alkaloids in S. flavescens could reduce Staphylococcus epidermidis tolerance to lactic acid. To sum up, Alkaloids in S. flavescens can be applied to the gynecological infection caused by biofilm-producing bacteria.
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OBJECTIVE: To study the effects of Kushen gel on the growth of six common vaginal lactobacilli in vitro.METHODS: This study was conducted at Beijing Tsinghua Changgung Hospital from June 2018 to March 2019.Six different vaginal lactobacilli isolated from the healthy women of reproductive age in China were purified and cultured in vitro. The effect of different concentrations of Sophora flavescens alkaloid,which was the main active ingredient of Kushen gel,on the growth of various clinical lactobacillus isolates was observed by the agar dilution method.RESULTS: The minimum inhibitory concentration(MIC)of Sophora flavescens alkaloid to Lactobacillus rhamnosus was 40 mg/mL. The MICs for Lactobacillus crispatus,Lactobacillus jensenii,Lactobacillus gasseri and Lactobacillus reuteri were all 20 mg/mL. The MIC for Lactobacillus vaginalis was 10 mg/mL.When the concentration of Sophora flavescens alkaloid was lower than the MIC value,it had a certain effect on promoting proliferation of both Lactobacillus gasseri and Lactobacillus reuteri,but had no effect on Lactobacillus crispatus or Lactobacillus vaginalis.CONCLUSION: Under the clinical dosage,Kushen gel does not inhibit the growth of common vaginal lactobacilli,which is conducive to the recovery of the vaginal microecosystem.
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In order to explore the chloroplast genome characteristics of Sophora flavescens and the phylogenetic relationship of the genus, this study used high-throughput sequencing technology to sequence and functionally annotate the chloroplast genome of Sophora flavescens. The results showed that the full length 154 165 bp of Sophora flavescens chloroplast genome showed a typical four-stage structure. The chloroplast genome contains 123 genes, including 77 protein-coding genes, 38 tRNA genes and 8 rRNA genes. Sequence analysis revealed 104 SSR loci, most of which consisted of A and T. In addition, the chloroplast genome codon preference is weak, and the coding region is biased towards the use of A and T bases. A comparative analysis of two different regions of Sophora flavescens chloroplast genome revealed four differential genes. Based on the maximum likelihood method (ML) for phylogenetic analysis of Sophora flavescens and 16 other leguminous, it was found that the relationship between Sophora flavescens and the genus Sophora alopecuroides is the closest. This study provides an important theoretical basis for the genetic variation, breeding and phylogenetic analysis of Sophora flavescens, and has certain reference value.
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Objective To analyze the clinical efficacy of Yanshu compound sophora flavescens injection (Yanshu injection) combined with paclitaxel and cisplatin (TP) regimen in the treatment of advanced non-small cell lung cancer (NSCLC). Methods One hundred and sixty-two patients with NSCLC admitted to the Department of Oncology of Danyang Hospital of Traditional Chinese Medicine from May 2014 to June 2018 were enrolled, all of their definite diagnosis was based on pathological or cytological examinations, and they were divided into two groups by the administration type, 81 cases in each group. The western medicine treatment group (western group) was treated with TP regimen alone, 3 weeks constituting 1 therapeutic course, and 3 consecutive courses were treated; based on the treatment of western drug TP regimen, the integrated traditional Chinese and western medicine treatment group (combined group) was additionally and simultaneously given Yanshu injection for consecutive 2 weeks. After treatment, the clinical efficacy and incidence of adverse reactions of the two groups were observed. Results The total effective rate in the combined group was significantly higher than that in western group [60.49% (49/81) vs. 40.74% (33/81), P <0.05]; the incidences of adverse events: alopecia, thrombocytopenia, neurotoxicity, leukopenia, nausea and vomiting, and gastrointestinal reaction in the combined group were obviously lower than those in western group [alopecia: 24.69% (20/81) vs. 60.49% (49/81), thrombocytopenia: 23.45% (19/81) vs. 40.74% (33/81), neurotoxicity: 14.81% (12/81) vs. 34.57% (28/81), leukopenia: 17.28% (14/81) vs. 62.96% (51/81), nausea and vomiting: 14.81% (12/81) vs. 60.49% (49/81), gastrointestinal reaction: 27.16% (22/81) vs. 62.96% (51/81), all P < 0.05]. Conclusion Yanshu injection combined with TP regimen is obviously effective in the treatment of advanced NSCLC, thus it is worthy to be applied clinically.
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To characterize the structure and function of ribosomal protein S13 (RPS13), we identified fulllength open reading frames (ORFs) of three RPS13 genes (RPS13-1, RPS13-2, and RPS13-3) of the Chinese medicinal plant, Sophora flavescens. The target genes were amplified by reverse transcription-olymerase chain reaction (RT-PCR), ligated into the pET22b(+) vector, and then transformed into Escherichia coli BL21 competent cells for protein expression. The physicochemical properties, protein motif, evolution, and structural organization of the three RPS13 genes were analyzed using bioinformatics tools. The full-length ORFs (453 bp) of the three RPS13 genes of S. flavescens were cloned, and each encodes a protein of 151 amino acids in length, and their expression was detected by Western blotting. Bioinformatics analysis showed that RPS13s are stable proteins that are closely related to the 40S RPS13s of Vigna radiate var. radiate. Their three-dimensional structures included three -helices at the C-terminal and four -helices at the N-terminal, and the two clusters of helices were connected by a long random coil, which may help maintain the dynamic bridging interactions between the large and small subunits of the ribosome. The full-length ORFs of three RPS13 genes of S. flavescens were successfully cloned and expressed in vitro. The study of the physicochemical properties, evolution, and secondary and three-dimensional structures of the three proteins will provide the theoretical basis for further studies on the function of RPS13s in plants.
Objetivo: Para caracterizar a estrutura e a função da proteína ribossomal S13 (RPS13), identificamos fases de leitura abertas (ORFs) completas de três genes RPS13 (RPS13-1, RPS13-2 e RPS13-3) da planta medicinal chinesa, Sophora flavescens. Métodos: Os genes alvo foram amplificados por reação em cadeia da polimerase por transcrição reversa (RT-PCR), ligados ao vetor pET22b(+), e então transformados em células competentes de Escherichia coli BL21 para expressão protéica. As propriedades físico-químicas, o motivo protéico, a evolução e a organização estrutural dos três genes RPS13 foram analisados utilizando ferramentas de bioinformática. Resultados: ORFs completos (453 pb) dos três genes RPS13 de S. flavescens foram clonados, e cada um codifica uma proteína de 151 aminoácidos de comprimento, e sua expressão foi detectada por western blotting. A análise de bioinformática mostrou que as RPS13s são proteínas estáveis que estão intimamente relacionadas com as 40S RPS13s de Vigna radiata var. radiate. Suas estruturas tridimensionais incluíam três -hélices no C-terminal e quatro -hélices no N-terminal, e os dois aglomerados de hélices eram conectados por uma longa bobina aleatória, o que pode ajudar a manter as interações de ponte dinâmicas entre o subunidades grandes e pequenas do ribossomo. Conclusões: As ORFs completas de três genes RPS13 de S. flavescens foram clonadas e expressas com sucesso in vitro. O estudo das propriedades físico-químicas, evolução e estruturas secundárias e tridimensionais das três proteínas fornecerão a base teórica para estudos adicionais sobre a função das RPS13s em plantas.
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Computational Biology , Sophora , Reverse Transcription , Escherichia coli , GenesABSTRACT
Objective To analyze the material basis and molecular mechanisms of Danggui Beimu Kushen (DBK) Pills in treating prostatic diseases based on the method of integrated pharmacology. Method The platform of Integrative Pharmacology of Traditional Chinese Medicine (TCM-IP, www.tcmip.cn) was utilized to predict the main active ingredients and functional targets of DBK Pills in treating prostatic disease, key targets were screened for enrichment analysis of pathways, and the network of “herb-core component-key target-main pathway” was constructed, and the possible mechanisms of DBK Pills in treating prostatic diseases were explored. Results A total of 532 candidate key targets for the treatment of prostatic diseases by DBK Pills were predicted, and 1 840 terms of gene function and 194 signal pathways were analyzed by gene ontology (GO) and KEGG, respectively. The network analysis of “herb-core component-key target-main pathway” showed that 65 core components were predicted, including 29 ingredients from Angelica sinensis, 11 from Fritillaria thunbergii and 26 from Sophora flavescens. Those predicted components acted on the key targets of prostatic diseases, such as transcription factor binding, negative regulation of apoptosis, et al, through the estrogen, apoptosis, chemokines and other signal pathways, and thus played a role in the regulation of cell cycle, apoptosis and proliferation imbalance, which might be the molecular mechanisms of DBK Pills for the treatment of prostatic disease. Conclusion DBK Pills regulate the development of BPH, prostate cancer and other diseases through multiple pathways with multi-component interacting with multiple targets.
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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
Objective:To develop an HPLC method for determining matrine , oxymatrine and oxysophocarpine in wine-processed Sophora flavescens.Methods:An HPLC method was used with a Dalian Elite HYP-ODS column (250 mm ×4.6 mm, 5 μm).The mobile phase consisted of methanol-phosphate buffer (12:88) with the flow rate of 1.0 ml· min-1 .The detection wavelength was 205 nm, the column temperature was 25℃and the injection volume was 20 μl.Results:The calibration curve was linear within the range of 0.105-0.420 μg for matrine,0.760-3.040μg for oxysophocarpine and 1.615-6.460μg for oxymatrine.The average recovery of ma-trine, oxysophocarpine and oxymatrine was 99.70％(RSD=1.92％), 99.41％( RSD=1.93％) and 98.60％(RSD=1.79％)(n=6), respectively.Conclusion:The method is accurate and reliable with good reproducibility , promising specificity and high sensitivi-ty, which can be used for the quality control of wine-processed Sophora flavescens.