该研究通过网络药理学的方法,对葵花护肝片活性成分作用靶点预测,探讨其多成分-多靶点-多通路肝脏保护作用模式。首先,采用中药系统药理学技术平台(TCMSP)及TCM Database@Taiwan资料库,以口服利用度(OB),类药性(DL)和药物半衰期(HL)筛...该研究通过网络药理学的方法,对葵花护肝片活性成分作用靶点预测,探讨其多成分-多靶点-多通路肝脏保护作用模式。首先,采用中药系统药理学技术平台(TCMSP)及TCM Database@Taiwan资料库,以口服利用度(OB),类药性(DL)和药物半衰期(HL)筛选葵花护肝片中活性成分。然后,依据反向药效团匹配(PharmMapper)方法预测葵花护肝片活性成分靶点;通过CooLGeN数据库文本挖掘及GeneCards数据库中与慢性肝炎及早期肝硬化相关靶基因比对筛选,借助生物信息学注释数据库(Metascape)分析靶点基因功能及信号通路,采用Cytoscape软件构建葵花护肝片化合物-靶点,化合物-靶点-通路。通过STRING数据库结合Cytoscape软件绘制蛋白相互作用(PPI)网络并进行网络拓扑学分析;最后,通过Systems Dock Web Site软件将葵花护肝片活性成分与枢纽靶点进行分子对接验证。实验筛选得到护肝片26个化合物和509个靶点;通过核心靶点蛋白互作网络分析确定了葵花护肝片活性成分与白蛋白(ALB)、胰岛素样生长因子1(IGF1)、基质金属蛋白酶-9 (MMP-9)、基质金属蛋白酶-2 (MMP-2)、非受体酪氨酸激酶原癌基因(SRC)及雌激素受体1(ESR1)等6个靶点及肿瘤-信号转导-炎症-药物代谢等相关生物过程和代谢通路紧密相关。结果表明,护肝片改善慢性肝炎及早期肝硬化作用体现了中药多成分-多靶点-多途径的作用特点,该研究为深入阐释葵花护肝片临床研究及进一步开发提供了思路。展开更多
Objective： To observe the effect of norcantharidin（NCTD） on collagen-induced arthritis（CIA） rats. Methods： Sixty Sprague-Dawley（SD） rats were randomly divided into 6 groups（n=10）： normal group, CIA model gr...Objective： To observe the effect of norcantharidin（NCTD） on collagen-induced arthritis（CIA） rats. Methods： Sixty Sprague-Dawley（SD） rats were randomly divided into 6 groups（n=10）： normal group, CIA model group（model group）, NCTD low-dose group [1.35 mg/（kg·d）], NCTD middle-dose group [2.7 mg/（kg·d）], NCTD high-dose group [5.4 mg/（kg·d）] and methotrexate（MTX） group [1.8 mg/（kg/w）]. Anesthetized rats were sacrificed by luxation of cervical vertebra after 4 weeks of administration. The arthritis scores were evaluated twice a week. The pathological changes in the ankle joints of rats were observed by hematoxylin-eosin（H＆E） staining. The serum levels of interleukin（IL） 1β, IL-6, tumor necrosis factor（TNF）-α, vascular endothelial growth factor（VEGF）, IL-17 and transform growth factor（TGF） β were detected by enzyme linked immunosorbent assay（ELISA）. The mRNA expression of retinoid-related orphan nuclear receptor γ t（ROR γ t） and forkhead box P3（Foxp3） in peripheral blood lymphocytes were confirmed by real-time polymerase chain reaction. Results： MTX and high-dose NCTD not only decreased the arthritis scores but also alleviated the pathological changes in CIA rats＇ ankle joints compared with the model group（P〈0.05 or P〈0.01）. All doses of NCTD significantly inhibited the serum levels of IL-6, IL-17 and TNF-α in CIA rats（P〈0.05）. Only middle-and high-dose of NCTD prominently decreased serum IL-1β and TGF-β levels of CIA rats（P〈0.05）. However, NCTD has no effect on vascular endothelial growth factor（VEGF） level in CIA rats. The Foxp3 mRNA expression in all NCTD groups were increased significantly than in the model group（P〈0.05）. The mRNA expression of RORγt in NCTD high-dose group was decreased apparently in comparison with the model group（P〈0.05）. Conclusion： NCTD showed therapeutic effect on CIA rats by inhibition of cytokines and regulation of Th17/Treg cells.展开更多
文摘该研究通过网络药理学的方法,对葵花护肝片活性成分作用靶点预测,探讨其多成分-多靶点-多通路肝脏保护作用模式。首先,采用中药系统药理学技术平台(TCMSP)及TCM Database@Taiwan资料库,以口服利用度(OB),类药性(DL)和药物半衰期(HL)筛选葵花护肝片中活性成分。然后,依据反向药效团匹配(PharmMapper)方法预测葵花护肝片活性成分靶点;通过CooLGeN数据库文本挖掘及GeneCards数据库中与慢性肝炎及早期肝硬化相关靶基因比对筛选,借助生物信息学注释数据库(Metascape)分析靶点基因功能及信号通路,采用Cytoscape软件构建葵花护肝片化合物-靶点,化合物-靶点-通路。通过STRING数据库结合Cytoscape软件绘制蛋白相互作用(PPI)网络并进行网络拓扑学分析;最后,通过Systems Dock Web Site软件将葵花护肝片活性成分与枢纽靶点进行分子对接验证。实验筛选得到护肝片26个化合物和509个靶点;通过核心靶点蛋白互作网络分析确定了葵花护肝片活性成分与白蛋白(ALB)、胰岛素样生长因子1(IGF1)、基质金属蛋白酶-9 (MMP-9)、基质金属蛋白酶-2 (MMP-2)、非受体酪氨酸激酶原癌基因(SRC)及雌激素受体1(ESR1)等6个靶点及肿瘤-信号转导-炎症-药物代谢等相关生物过程和代谢通路紧密相关。结果表明,护肝片改善慢性肝炎及早期肝硬化作用体现了中药多成分-多靶点-多途径的作用特点,该研究为深入阐释葵花护肝片临床研究及进一步开发提供了思路。
基金Supported by the National Natural Science Foundation of China(No.81273837)
文摘Objective： To observe the effect of norcantharidin（NCTD） on collagen-induced arthritis（CIA） rats. Methods： Sixty Sprague-Dawley（SD） rats were randomly divided into 6 groups（n=10）： normal group, CIA model group（model group）, NCTD low-dose group [1.35 mg/（kg·d）], NCTD middle-dose group [2.7 mg/（kg·d）], NCTD high-dose group [5.4 mg/（kg·d）] and methotrexate（MTX） group [1.8 mg/（kg/w）]. Anesthetized rats were sacrificed by luxation of cervical vertebra after 4 weeks of administration. The arthritis scores were evaluated twice a week. The pathological changes in the ankle joints of rats were observed by hematoxylin-eosin（H＆E） staining. The serum levels of interleukin（IL） 1β, IL-6, tumor necrosis factor（TNF）-α, vascular endothelial growth factor（VEGF）, IL-17 and transform growth factor（TGF） β were detected by enzyme linked immunosorbent assay（ELISA）. The mRNA expression of retinoid-related orphan nuclear receptor γ t（ROR γ t） and forkhead box P3（Foxp3） in peripheral blood lymphocytes were confirmed by real-time polymerase chain reaction. Results： MTX and high-dose NCTD not only decreased the arthritis scores but also alleviated the pathological changes in CIA rats＇ ankle joints compared with the model group（P〈0.05 or P〈0.01）. All doses of NCTD significantly inhibited the serum levels of IL-6, IL-17 and TNF-α in CIA rats（P〈0.05）. Only middle-and high-dose of NCTD prominently decreased serum IL-1β and TGF-β levels of CIA rats（P〈0.05）. However, NCTD has no effect on vascular endothelial growth factor（VEGF） level in CIA rats. The Foxp3 mRNA expression in all NCTD groups were increased significantly than in the model group（P〈0.05）. The mRNA expression of RORγt in NCTD high-dose group was decreased apparently in comparison with the model group（P〈0.05）. Conclusion： NCTD showed therapeutic effect on CIA rats by inhibition of cytokines and regulation of Th17/Treg cells.
