Comprehensive characterization of metabolites and metabolic profiles in plasma has considerable significance in determining the efficacy and safety of traditional Chinese medicine(TCM)in vivo.However,this process is u...Comprehensive characterization of metabolites and metabolic profiles in plasma has considerable significance in determining the efficacy and safety of traditional Chinese medicine(TCM)in vivo.However,this process is usually hindered by the insufficient characteristic fragments of metabolites,ubiquitous matrix interference,and complicated screening and identification procedures for metabolites.In this study,an effective strategy was established to systematically characterize the metabolites,deduce the metabolic pathways,and describe the metabolic profiles of bufadienolides isolated from Venenum Bufonis in vivo.The strategy was divided into five steps.First,the blank and test plasma samples were injected into an ultra-high performance liquid chromatography/linear trap quadrupole-orbitrap-mass spectrometry(MS)system in the full scan mode continuously five times to screen for valid matrix compounds and metabolites.Second,an extension-mass defect filter model was established to obtain the targeted precursor ions of the list of bufadienolide metabolites,which reduced approximately 39%of the interfering ions.Third,an acquisition model was developed and used to trigger more tandem MS(MS/MS)fragments of precursor ions based on the targeted ion list.The acquisition mode enhanced the acquisition capability by approximately four times than that of the regular data-dependent acquisition mode.Fourth,the acquired data were imported into Compound Discoverer software for identification of metabolites with metabolic network prediction.The main in vivo metabolic pathways of bufadienolides were elucidated.A total of 147 metabolites were characterized,and the main biotransformation reactions of bufadienolides were hydroxylation,dihydroxylation,and isomerization.Finally,the main prototype bufadienolides in plasma at different time points were determined using LC-MS/MS,and the metabolic profiles were clearly identified.This strategy could be widely used to elucidate the metabolic profiles of TCM preparations or Chinese patent medicines in vivo and provide critical data for rational drug use.展开更多
A high performance liquid chromatographic method was used for the simultaneous identification and qualitative evaluation of 12 bufadienolides(resibufogenin, cinobufagin, cinobufaginol, arenobufagin, bufalin, bufotalin...A high performance liquid chromatographic method was used for the simultaneous identification and qualitative evaluation of 12 bufadienolides(resibufogenin, cinobufagin, cinobufaginol, arenobufagin, bufalin, bufotalin, gamabufotalin, cinobufotalin, Ψ-bufaranogin, desacetylcinobufagin, telocinobufagin and resibufogenol) in Venenum Bufonis. The chromatographic separation was performed on a Dikma C18 analytical column via gradient elution with an aqueous solution of acetonitrile and 0.3% acetic acid at a flow rate of 0.8 mL/min. The method was validated to be acceptable in consideration of linearity(r2>0.9992) and recovery(ranged from 98.9% to 102.0%). The limits of detection of the bufadienolides were from 0.48 ng for bufalin to 6.00 ng for cinobufotalin. The intra-day and inter-day precisions of the method were evaluated and were less than 3.0%. The method was successfully used to analyze 19 batches of Venenum Bufonis, and the similarity values between batches were calculated by Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version 2004A, Chinese Pharmacopoeia Committee, Beijing). The results show that the contents of bufadienolides in the medicine and the similarity values based on these bufadieno-lides varied significantly from batch to batch. This proposed method could be utilized to qualify and control Venenum Bufonis to ensure its safety and efficacy in application.展开更多
Ethanol extract of Venenum Bufonis has shown many valuable bioactivities, but little is known about its effect on biofilm formation of Staphylococcus aureus. In this study, biofilm formation of S. aureus NCTC8325 with...Ethanol extract of Venenum Bufonis has shown many valuable bioactivities, but little is known about its effect on biofilm formation of Staphylococcus aureus. In this study, biofilm formation of S. aureus NCTC8325 with or without ethanol extract of Venenum Bufonis was tested using microtitre plate assay and Confocal Laser Scanning Microscope (CLSM) system. Results show that the biofilm formation of S. aureus with ethanol extract of Venenum Bufonis was significantly lower than that of the control group without ethanol extract of Venenum Bufonis. Meanwhile, the reduction degree was correlated to the concentration of ethanol extract of Venenum Bufonis positively. With CLSM system we can observe that looser and less biomass of the biofilm structure of the experimental group appeared than that of the control group. These results suggested that ethanol extract of Venenum Bufonis has inhibitory effect on biofilm formation of S. aureus.展开更多
Venenum Bufonis is one of the traditional medical herbs,containing a variety of active ingredients such as peptides,alkaloids,lactones and sterols.In recent years,the significant anti-tumor effect of Venenum Bufonis a...Venenum Bufonis is one of the traditional medical herbs,containing a variety of active ingredients such as peptides,alkaloids,lactones and sterols.In recent years,the significant anti-tumor effect of Venenum Bufonis and its active components,which are mainly composed of lipid scorpion venom,scorpion venom and scorpion venom,has become a research hotspot.Venenum Bufonis can inhibit the occurrence and development of tumor cells by inducing apoptosis of tumor cells,promoting tumor cell differentiation,inhibiting tumor microangiogenesis and reversing tumor cell resistance.Based on the chemical composition of Venenum Bufonis,this paper summarizes the mechanism of inhibiting tumors of the active ingredients in Venenum Bufonis.展开更多
检索了中国知网及Web of Science数据库近10年发表的蟾酥相关的中、英文文献,应用文献计量学和CiteSpace 5.8.R3可视化软件对作者、研究机构、关键词及文献来源等进行分析,并结合信息提取方法对蟾酥研究的现状及发展趋势进行分析。经过...检索了中国知网及Web of Science数据库近10年发表的蟾酥相关的中、英文文献,应用文献计量学和CiteSpace 5.8.R3可视化软件对作者、研究机构、关键词及文献来源等进行分析,并结合信息提取方法对蟾酥研究的现状及发展趋势进行分析。经过筛选最终纳入447篇中文文献和316篇英文文献。叶文才是该领域发表文献最多的作者;山东中医药大学、暨南大学分别是发表蟾酥相关中、英文文献量最多的机构;中文领域机构间合作有待提升,英文领域机构间的合作较为普遍。分析中、英文关键词,发现蟾酥相关研究主要集中在有效成分、安全性、质量标准等方面,抗肿瘤机制和毒效成分研究是目前及未来的热点。展开更多
基金supported by the National Natural Science Foundation of China (Grant Nos.: 81530095 and 81673591)Strategic Priority Research Program of the Chinese Academy of Sciences (Grant No.: XDA12020348)+1 种基金National Standardization of Traditional Chinese Medicine Project (Grant No.: ZYBZH-K-LN-01)Science and Technology Commission Foundation of Shanghai (Grant No.: 15DZ0502800)
文摘Comprehensive characterization of metabolites and metabolic profiles in plasma has considerable significance in determining the efficacy and safety of traditional Chinese medicine(TCM)in vivo.However,this process is usually hindered by the insufficient characteristic fragments of metabolites,ubiquitous matrix interference,and complicated screening and identification procedures for metabolites.In this study,an effective strategy was established to systematically characterize the metabolites,deduce the metabolic pathways,and describe the metabolic profiles of bufadienolides isolated from Venenum Bufonis in vivo.The strategy was divided into five steps.First,the blank and test plasma samples were injected into an ultra-high performance liquid chromatography/linear trap quadrupole-orbitrap-mass spectrometry(MS)system in the full scan mode continuously five times to screen for valid matrix compounds and metabolites.Second,an extension-mass defect filter model was established to obtain the targeted precursor ions of the list of bufadienolide metabolites,which reduced approximately 39%of the interfering ions.Third,an acquisition model was developed and used to trigger more tandem MS(MS/MS)fragments of precursor ions based on the targeted ion list.The acquisition mode enhanced the acquisition capability by approximately four times than that of the regular data-dependent acquisition mode.Fourth,the acquired data were imported into Compound Discoverer software for identification of metabolites with metabolic network prediction.The main in vivo metabolic pathways of bufadienolides were elucidated.A total of 147 metabolites were characterized,and the main biotransformation reactions of bufadienolides were hydroxylation,dihydroxylation,and isomerization.Finally,the main prototype bufadienolides in plasma at different time points were determined using LC-MS/MS,and the metabolic profiles were clearly identified.This strategy could be widely used to elucidate the metabolic profiles of TCM preparations or Chinese patent medicines in vivo and provide critical data for rational drug use.
基金Supported by the National Basic Researeh Program of China(No.2007CB507400)the National High Technology Research and Development Program(No.2006AA02Z338)+1 种基金Program for Changjiang Scholars and Innovative Research Team in Universities of China(PCSIRT)Partly Supported by the Scientific Foundation of Shanghai City,China(Nos.05DZ19733,06DZ19717,06DZ19005)
文摘A high performance liquid chromatographic method was used for the simultaneous identification and qualitative evaluation of 12 bufadienolides(resibufogenin, cinobufagin, cinobufaginol, arenobufagin, bufalin, bufotalin, gamabufotalin, cinobufotalin, Ψ-bufaranogin, desacetylcinobufagin, telocinobufagin and resibufogenol) in Venenum Bufonis. The chromatographic separation was performed on a Dikma C18 analytical column via gradient elution with an aqueous solution of acetonitrile and 0.3% acetic acid at a flow rate of 0.8 mL/min. The method was validated to be acceptable in consideration of linearity(r2>0.9992) and recovery(ranged from 98.9% to 102.0%). The limits of detection of the bufadienolides were from 0.48 ng for bufalin to 6.00 ng for cinobufotalin. The intra-day and inter-day precisions of the method were evaluated and were less than 3.0%. The method was successfully used to analyze 19 batches of Venenum Bufonis, and the similarity values between batches were calculated by Similarity Evaluation System for Chromatographic Fingerprint of TCM(Version 2004A, Chinese Pharmacopoeia Committee, Beijing). The results show that the contents of bufadienolides in the medicine and the similarity values based on these bufadieno-lides varied significantly from batch to batch. This proposed method could be utilized to qualify and control Venenum Bufonis to ensure its safety and efficacy in application.
文摘Ethanol extract of Venenum Bufonis has shown many valuable bioactivities, but little is known about its effect on biofilm formation of Staphylococcus aureus. In this study, biofilm formation of S. aureus NCTC8325 with or without ethanol extract of Venenum Bufonis was tested using microtitre plate assay and Confocal Laser Scanning Microscope (CLSM) system. Results show that the biofilm formation of S. aureus with ethanol extract of Venenum Bufonis was significantly lower than that of the control group without ethanol extract of Venenum Bufonis. Meanwhile, the reduction degree was correlated to the concentration of ethanol extract of Venenum Bufonis positively. With CLSM system we can observe that looser and less biomass of the biofilm structure of the experimental group appeared than that of the control group. These results suggested that ethanol extract of Venenum Bufonis has inhibitory effect on biofilm formation of S. aureus.
文摘Venenum Bufonis is one of the traditional medical herbs,containing a variety of active ingredients such as peptides,alkaloids,lactones and sterols.In recent years,the significant anti-tumor effect of Venenum Bufonis and its active components,which are mainly composed of lipid scorpion venom,scorpion venom and scorpion venom,has become a research hotspot.Venenum Bufonis can inhibit the occurrence and development of tumor cells by inducing apoptosis of tumor cells,promoting tumor cell differentiation,inhibiting tumor microangiogenesis and reversing tumor cell resistance.Based on the chemical composition of Venenum Bufonis,this paper summarizes the mechanism of inhibiting tumors of the active ingredients in Venenum Bufonis.
文摘检索了中国知网及Web of Science数据库近10年发表的蟾酥相关的中、英文文献,应用文献计量学和CiteSpace 5.8.R3可视化软件对作者、研究机构、关键词及文献来源等进行分析,并结合信息提取方法对蟾酥研究的现状及发展趋势进行分析。经过筛选最终纳入447篇中文文献和316篇英文文献。叶文才是该领域发表文献最多的作者;山东中医药大学、暨南大学分别是发表蟾酥相关中、英文文献量最多的机构;中文领域机构间合作有待提升,英文领域机构间的合作较为普遍。分析中、英文关键词,发现蟾酥相关研究主要集中在有效成分、安全性、质量标准等方面,抗肿瘤机制和毒效成分研究是目前及未来的热点。
