According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloid...According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloids in mother roots, fibrous roots, stems, and leaves of Aconitum carmichaelii Debx. were measured by HPLC-MS/MS. And multivariate analysis methods, such as clustering analysis and principal component analysis, were applied to analyze the difference among various parts. In addition, the acute toxicity, analgesia, and anti-inflammatory tests were carried out. The results suggested that the contents of alkaloids in mother roots and fibrous roots were approximate, but those of leaves and stems were different from mother roots and fibrous roots. The results of the acute toxicity testing demonstrated the toxicity of fibrous root was strongest, and mother roots were slightly less toxic than fibrous roots. The stems and leaves were far less toxic than mother and fibrous roots. In addition, the analgesia and inflammatory tests showed the effects of the various tissues had no difference each other. These results provided a basis for developing new complementary and alternative treatments for rheumatoid arthritis patients. Simultaneously, the approach may also turn wastes into treasure and promote the development of circular economy.展开更多
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
Objective:To investigate the effect and the mechanism of Astragalus membranaceus(Huangqi in Chinese,HQ)extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii(Fuzi in Chinese,FZ)in rats with spl...Objective:To investigate the effect and the mechanism of Astragalus membranaceus(Huangqi in Chinese,HQ)extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii(Fuzi in Chinese,FZ)in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier.Methods:Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d.Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids.Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins(TJ,including Claudin-1,Occludin and ZO-1)were measured using Western blot and real-time PCR,respectively.The location and expression of TJ protein was also investigated by the immunofluorescence method.Results:Compared with the normal group,the protein expression of MRP2,BCRP and TJ proteins in the model group were significantly down-regulated.After oral administration of HQ,the alkaloid absorption in intestinal villi was inhibited,MRP2,BCRP and TJ proteins were up-regulated,the green fluorescence staining of Claudin-1,Occludin,and ZO-1 was enhanced,and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity.Conclusion:HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components,leading to a decrease in the absorption of drug-like molecules.展开更多
目的基于网络药理学及分子对接技术研究川乌、草乌相须配伍治疗类风湿性关节炎潜在的有效成分及其作用机制,为川乌、草乌治疗类风湿性关节炎提供参考。方法通过中药系统药理学数据库及分析平台(TCMSP)中以口服生物利用度(oral bioavaila...目的基于网络药理学及分子对接技术研究川乌、草乌相须配伍治疗类风湿性关节炎潜在的有效成分及其作用机制,为川乌、草乌治疗类风湿性关节炎提供参考。方法通过中药系统药理学数据库及分析平台(TCMSP)中以口服生物利用度(oral bioavailability,OB)、类药性(Drug-likeness,DL)参数筛选川乌、草乌的成分,采用PharmMapper数据库寻找川乌、草乌的作用靶点,与疾病数据库GeneCards及OMIM数据库中类风湿性关节炎的靶点取交集。在String平台进行蛋白质-蛋白质相互作用(PPI)分析,Metascape平台进行基因本体(Gene Ontology,GO)分析及京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。通过Cytoscape软件构建“药物-成分-类风湿性关节炎靶点-通路”网络。利用SYBYL软件进行分子对接。结果筛选出10个潜在活性成分(次乌头碱、滇乌头碱、草乌甲素等),23个对应治疗靶点(TGFB2、IL2、MMP3、MMP2等),涉及类风湿性关节炎、白细胞介素-17、MAPK、JAK-STAT等通路及炎症反应、防御反应等生物过程。结论川乌、草乌通过多成分、多靶点、多通路协同治疗类风湿性关节炎,且有多个成分表现出很强的活性,为后续研究提供一定的参考。展开更多
基金supported by the Sichuan Provincial Administration of Traditional Chinese Medicine Research Project(No.2018NQ008)
文摘According to folk usage of Aconitum carmichaelii Debx., the present study was designed to determine the feasibility of the stems and leaves of Aconitum carmichaelii Debx. as a new medicinal resource. Fourteen alkaloids in mother roots, fibrous roots, stems, and leaves of Aconitum carmichaelii Debx. were measured by HPLC-MS/MS. And multivariate analysis methods, such as clustering analysis and principal component analysis, were applied to analyze the difference among various parts. In addition, the acute toxicity, analgesia, and anti-inflammatory tests were carried out. The results suggested that the contents of alkaloids in mother roots and fibrous roots were approximate, but those of leaves and stems were different from mother roots and fibrous roots. The results of the acute toxicity testing demonstrated the toxicity of fibrous root was strongest, and mother roots were slightly less toxic than fibrous roots. The stems and leaves were far less toxic than mother and fibrous roots. In addition, the analgesia and inflammatory tests showed the effects of the various tissues had no difference each other. These results provided a basis for developing new complementary and alternative treatments for rheumatoid arthritis patients. Simultaneously, the approach may also turn wastes into treasure and promote the development of circular economy.
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。
基金supported by a grant from the Key Laboratory of Modern Preparation of TCM of Ministry of Education,Jiangxi University of TCM and Affiliated Hospital,Jiujiang UniversityScience Foundation of Health and Family Planning Commission of Jiangxi Province (Grant No.20181140)+2 种基金National Natural Science Foundation of China (Grant No.81660757)Project of Jiangxi Education Department (Grant No.170732Grant No.201819)
文摘Objective:To investigate the effect and the mechanism of Astragalus membranaceus(Huangqi in Chinese,HQ)extract on the intestinal absorption of six alkaloids of Aconitum carmichaelii(Fuzi in Chinese,FZ)in rats with spleen deficiency and provide novel insights into the application of HQ on modulating intestinal barrier.Methods:Four-week-old male Sprague-Dawley rats were fed with Xiaochengqi Decoction to induce the spleen deficiency model for 40 d.Single-pass intestinal perfusion model were used to study the effects of HQ extract on the absorption of alkaloids.Protein expression and mRNA levels of MRP2 and BCRP and tight junction proteins(TJ,including Claudin-1,Occludin and ZO-1)were measured using Western blot and real-time PCR,respectively.The location and expression of TJ protein was also investigated by the immunofluorescence method.Results:Compared with the normal group,the protein expression of MRP2,BCRP and TJ proteins in the model group were significantly down-regulated.After oral administration of HQ,the alkaloid absorption in intestinal villi was inhibited,MRP2,BCRP and TJ proteins were up-regulated,the green fluorescence staining of Claudin-1,Occludin,and ZO-1 was enhanced,and a thick layer of mucus was deposited on the surface of the epithelium of the intestinal cavity.Conclusion:HQ as an intestinal barrier modulator improves the physiological changes of the intestinal environment of spleen deficiency to reduce the absorption of toxic components,leading to a decrease in the absorption of drug-like molecules.
文摘目的基于网络药理学及分子对接技术研究川乌、草乌相须配伍治疗类风湿性关节炎潜在的有效成分及其作用机制,为川乌、草乌治疗类风湿性关节炎提供参考。方法通过中药系统药理学数据库及分析平台(TCMSP)中以口服生物利用度(oral bioavailability,OB)、类药性(Drug-likeness,DL)参数筛选川乌、草乌的成分,采用PharmMapper数据库寻找川乌、草乌的作用靶点,与疾病数据库GeneCards及OMIM数据库中类风湿性关节炎的靶点取交集。在String平台进行蛋白质-蛋白质相互作用(PPI)分析,Metascape平台进行基因本体(Gene Ontology,GO)分析及京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析。通过Cytoscape软件构建“药物-成分-类风湿性关节炎靶点-通路”网络。利用SYBYL软件进行分子对接。结果筛选出10个潜在活性成分(次乌头碱、滇乌头碱、草乌甲素等),23个对应治疗靶点(TGFB2、IL2、MMP3、MMP2等),涉及类风湿性关节炎、白细胞介素-17、MAPK、JAK-STAT等通路及炎症反应、防御反应等生物过程。结论川乌、草乌通过多成分、多靶点、多通路协同治疗类风湿性关节炎,且有多个成分表现出很强的活性,为后续研究提供一定的参考。
