目的运用网络药理学和体外细胞实验探究附子活性成分治疗血管痉挛的潜在作用机制。方法检索TCMSP数据库、SwissTargetPrediction数据库、Uniprot数据库、Genecards数据库、OMIM数据库、TTD数据库、Disgenet数据库和String数据库等,获得...目的运用网络药理学和体外细胞实验探究附子活性成分治疗血管痉挛的潜在作用机制。方法检索TCMSP数据库、SwissTargetPrediction数据库、Uniprot数据库、Genecards数据库、OMIM数据库、TTD数据库、Disgenet数据库和String数据库等,获得附子活性成分、作用靶点及血管痉挛相应疾病靶点,并将其作用靶点与获得的疾病靶点的交集作为潜在靶点;利用String平台,对潜在靶点构建蛋白—蛋白互作(protein-protein interaction network,PPI)网络;采用R语言软件包对潜在靶点进行基因本体(gene ontology,GO)功能及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析,然后使用Cytoscape软件分析其网络拓扑结构,筛选其核心靶点,并进行分子对接验证。在此基础上,通过MTT法检测附子活性成分对血管紧张素Ⅱ所致血管平滑肌细胞增殖影响的机制。结果附子治疗血管痉挛的主要潜在活性成分有15个;药物—疾病核心靶点有27个;涉及的信号通路主要有脂质和动脉粥样硬化、钙信号通路、刺激神经组织的中的交互、鞘脂类信号通路、流体剪切应力与动脉粥样硬化、松弛素信号通路、血管平滑肌收缩等;关键核心靶点与活性成分次乌头碱、谷甾醇结合具有稳定构象。体外实验结果表明,附子活性成分次乌头碱能抑制血管紧张素Ⅱ所致血管平滑肌细胞增殖,其机制可能与其抑制丝裂原活化蛋白激酶信号转导通路的活化有关。结论附子多种活性成分可能通过作用于丝裂原活化蛋白激酶1、核受体亚家族3 C群成员1、血管紧张素I转化酶、一氧化氮合酶3等靶点治疗血管痉挛,为附子活性成分治疗血管痉挛的作用机制研究提供了科学参考。展开更多
A new aporphine alkaloid, named fuzitine, was isolated from the tuber of Aconitum carmicheali Debx, and its chemical structure was established'on the basis of spectral analysis and chemical reaction.
In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after co...In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after codecocting of the combination of Aconitum carmichaeli Debx.(wu-tou in Chinese, WT) and Bletilla striata(Thunb.) Reichb.f.(bai-ji in Chinese, BJ) incompatible pair. Two different kinds of decoctions, namely WT-BJ mixed decoction: mixed water extract of each individual herbs, and WT-BJ co-decoction: water extract of mixed two constituent herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-QTOF/MS analysis, the datasets of tR-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis(OPLS-DA) to holistically compare the difference between these two kinds of decoction samples. Once a clear classification trend was found in score plot, extended statistical analysis was performed to generate S-plot, in which the variables(tR-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components that correlate to these ions were regarded as the most changed components during co-decocting of the incompatible pair. The identities of the changed components can be identified by comparing the retention times and mass spectra with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literatures. Using the proposed approach, global chemical difference was found between mixed decoction and co-decoction, and hypaconitine, mesaconitine, deoxyaconitine, aconitine, 10-OH-mesaconitine, 10-OH-aconitine and deoxyhypaconitine were identified as the most changed toxic components of the combination of WT-BJ incompatible pair during co-decocting. It is suggested that this newly established approach could be used to practically reveal the possible toxic components changed/increased of the herbal combination taboos, e.g. the Eighteen Incompatible Medications(Shi Ba Fan), in traditional Chinese medicines.展开更多
文摘目的运用网络药理学和体外细胞实验探究附子活性成分治疗血管痉挛的潜在作用机制。方法检索TCMSP数据库、SwissTargetPrediction数据库、Uniprot数据库、Genecards数据库、OMIM数据库、TTD数据库、Disgenet数据库和String数据库等,获得附子活性成分、作用靶点及血管痉挛相应疾病靶点,并将其作用靶点与获得的疾病靶点的交集作为潜在靶点;利用String平台,对潜在靶点构建蛋白—蛋白互作(protein-protein interaction network,PPI)网络;采用R语言软件包对潜在靶点进行基因本体(gene ontology,GO)功能及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析,然后使用Cytoscape软件分析其网络拓扑结构,筛选其核心靶点,并进行分子对接验证。在此基础上,通过MTT法检测附子活性成分对血管紧张素Ⅱ所致血管平滑肌细胞增殖影响的机制。结果附子治疗血管痉挛的主要潜在活性成分有15个;药物—疾病核心靶点有27个;涉及的信号通路主要有脂质和动脉粥样硬化、钙信号通路、刺激神经组织的中的交互、鞘脂类信号通路、流体剪切应力与动脉粥样硬化、松弛素信号通路、血管平滑肌收缩等;关键核心靶点与活性成分次乌头碱、谷甾醇结合具有稳定构象。体外实验结果表明,附子活性成分次乌头碱能抑制血管紧张素Ⅱ所致血管平滑肌细胞增殖,其机制可能与其抑制丝裂原活化蛋白激酶信号转导通路的活化有关。结论附子多种活性成分可能通过作用于丝裂原活化蛋白激酶1、核受体亚家族3 C群成员1、血管紧张素I转化酶、一氧化氮合酶3等靶点治疗血管痉挛,为附子活性成分治疗血管痉挛的作用机制研究提供了科学参考。
文摘A new aporphine alkaloid, named fuzitine, was isolated from the tuber of Aconitum carmicheali Debx, and its chemical structure was established'on the basis of spectral analysis and chemical reaction.
基金supported by the National Basic Research Program of China("973Program)(No2011CB505304)the Youth Scientific Research Project of Anhui Academy of Medical Science(YKY2018003)
文摘In the present study, an ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS) based chemical profiling approach to rapidly evaluate chemical diversity after codecocting of the combination of Aconitum carmichaeli Debx.(wu-tou in Chinese, WT) and Bletilla striata(Thunb.) Reichb.f.(bai-ji in Chinese, BJ) incompatible pair. Two different kinds of decoctions, namely WT-BJ mixed decoction: mixed water extract of each individual herbs, and WT-BJ co-decoction: water extract of mixed two constituent herbs, were prepared. Batches of these two kinds of decoction samples were subjected to UPLC-QTOF/MS analysis, the datasets of tR-m/z pairs, ion intensities and sample codes were processed with supervised orthogonal partial least squared discriminant analysis(OPLS-DA) to holistically compare the difference between these two kinds of decoction samples. Once a clear classification trend was found in score plot, extended statistical analysis was performed to generate S-plot, in which the variables(tR-m/z pair) contributing most to the difference were clearly depicted as points at the two ends of "S", and the components that correlate to these ions were regarded as the most changed components during co-decocting of the incompatible pair. The identities of the changed components can be identified by comparing the retention times and mass spectra with those of reference compounds and/or tentatively assigned by matching empirical molecular formulae with those of the known compounds published in the literatures. Using the proposed approach, global chemical difference was found between mixed decoction and co-decoction, and hypaconitine, mesaconitine, deoxyaconitine, aconitine, 10-OH-mesaconitine, 10-OH-aconitine and deoxyhypaconitine were identified as the most changed toxic components of the combination of WT-BJ incompatible pair during co-decocting. It is suggested that this newly established approach could be used to practically reveal the possible toxic components changed/increased of the herbal combination taboos, e.g. the Eighteen Incompatible Medications(Shi Ba Fan), in traditional Chinese medicines.
