通过网络药理学方法探讨逍遥散对动脉粥样硬化和抑郁症的"异病同治"的作用机制。通过TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform)、SymMap数据库检索逍遥散中8味中药相关的所...通过网络药理学方法探讨逍遥散对动脉粥样硬化和抑郁症的"异病同治"的作用机制。通过TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform)、SymMap数据库检索逍遥散中8味中药相关的所有化学成分和作用靶点,经初步筛选后构建"中药-化合物-靶点"作用网络,通过DisGeNET、CTD(Comparative Toxicogenomics Database)和TTD(Therapeutic Target Database)数据库获取动脉粥样硬化和抑郁症的相关基因,将药物作用靶点和疾病基因靶点整合获得共有靶点,使用STRING 11.0和Cytoscape进行共有靶点的蛋白质间相互作用分析筛选关键共有靶点,应用BioGPS获取其在器官组织的分布信息,并使用Metascape对关键共有靶点进行基因本体(gene ontology,GO)分析、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析。结果显示,检索到逍遥散化合物共1355个,据口服生物利用度≥30%和类药性指数≥0.18标准筛选出161个活性化合物,获得274个药物作用靶点,构建"中药-化合物-靶点"作用网络;检索到动脉粥样硬化疾病基因靶点共1004个,抑郁症578个,通过与药物作用靶点取交集后获得37个共有靶点;使用STRING和Cytoscape进行共有靶点的蛋白质间相互作用分析筛选关键共有靶点18个,BioGPS显示关键共有靶点主要分布在心脏、杏仁核、松果体、肝脏、平滑肌中;Metascape进行GO功能富集分析得到生物过程929个、细胞组成25个、分子功能23个,KEGG富集分析获得108条信号通路,靠前排序的信号通路如AGE-RAGE,HIF-1,FoxO,Th17细胞分化和IL-17信号通路,主要与神经内分泌、代谢、免疫炎症以及氧化应激相关。该研究表明,逍遥散"异病同治"动脉粥样硬化和抑郁症的主要机制涉及神经内分泌、代谢、免疫炎症以及氧化应激相关信号通路,为进一步试验验证、潜在药理学机制及临床拓展应用提供参考依据。展开更多
Objective: To test the hypothesis that the inhibition of endoplasmic reticulum(ER) stress-induced apoptosis in oxidized low-density lipoproteins(ox-LDL)-induced human aortic-vascular smooth muscle cells(HA-VSMCs) was ...Objective: To test the hypothesis that the inhibition of endoplasmic reticulum(ER) stress-induced apoptosis in oxidized low-density lipoproteins(ox-LDL)-induced human aortic-vascular smooth muscle cells(HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase(PERK)-eukaryotic translation initiation factor 2α(e IF2α)-activating transcription factor 4(ATF4)-CCAAT/enhancer binding protein homologous protein(CHOP) signaling pathway by Pollen Typhae total flavone(PTF). Methods: Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group(70 μg/m L high ox-LDL), an HPTF group(70 μg/m L high ox-LDL+500 μg/m L PTF), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), and a LPTF group(70 μg/m L high ox-LDL+100 μg/m L PTF) in the first part;and a normal control group, an ox-LDL group(70 μg/mL high ox-LDL), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), a sh RNA group(transducted with PERK shRNA lentiviral particles), a scramble shRNA group(transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group(250 μg/mL PTF+70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group(70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their m RNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was applied to test cel viability, and the level of apoptosis was monitored by flow cytometry. Results: The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and sh RNA groups. Moreover, the ox-LDL group had increased protein and m RNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK, e IF2α, ATF4 and CHOP, which were attenuated in PTF treatment groups and sh RNA groups. Conclusions: The apoptosis induced by ox-LDL had a strong relation to ER stress. The protective effect of PTF on ER stressinduced apoptosis was associated with inhibition of the PERK-eIF2α-ATF4-CHOP pathway, which might be a potential therapeutic strategy for enhancing the stability of atherosclerotic plaques.展开更多
文摘通过网络药理学方法探讨逍遥散对动脉粥样硬化和抑郁症的"异病同治"的作用机制。通过TCMSP(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform)、SymMap数据库检索逍遥散中8味中药相关的所有化学成分和作用靶点,经初步筛选后构建"中药-化合物-靶点"作用网络,通过DisGeNET、CTD(Comparative Toxicogenomics Database)和TTD(Therapeutic Target Database)数据库获取动脉粥样硬化和抑郁症的相关基因,将药物作用靶点和疾病基因靶点整合获得共有靶点,使用STRING 11.0和Cytoscape进行共有靶点的蛋白质间相互作用分析筛选关键共有靶点,应用BioGPS获取其在器官组织的分布信息,并使用Metascape对关键共有靶点进行基因本体(gene ontology,GO)分析、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)分析。结果显示,检索到逍遥散化合物共1355个,据口服生物利用度≥30%和类药性指数≥0.18标准筛选出161个活性化合物,获得274个药物作用靶点,构建"中药-化合物-靶点"作用网络;检索到动脉粥样硬化疾病基因靶点共1004个,抑郁症578个,通过与药物作用靶点取交集后获得37个共有靶点;使用STRING和Cytoscape进行共有靶点的蛋白质间相互作用分析筛选关键共有靶点18个,BioGPS显示关键共有靶点主要分布在心脏、杏仁核、松果体、肝脏、平滑肌中;Metascape进行GO功能富集分析得到生物过程929个、细胞组成25个、分子功能23个,KEGG富集分析获得108条信号通路,靠前排序的信号通路如AGE-RAGE,HIF-1,FoxO,Th17细胞分化和IL-17信号通路,主要与神经内分泌、代谢、免疫炎症以及氧化应激相关。该研究表明,逍遥散"异病同治"动脉粥样硬化和抑郁症的主要机制涉及神经内分泌、代谢、免疫炎症以及氧化应激相关信号通路,为进一步试验验证、潜在药理学机制及临床拓展应用提供参考依据。
基金Supported by the National Natural Science Foundation of China(No.81573922)the Traditional Chinese Medicine Scientific Research Project of Guangdong Province,China(No.20151076)the Sanming Project of Medicine in Shenzhen,China(No.SZSM201612033)
文摘Objective: To test the hypothesis that the inhibition of endoplasmic reticulum(ER) stress-induced apoptosis in oxidized low-density lipoproteins(ox-LDL)-induced human aortic-vascular smooth muscle cells(HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase(PERK)-eukaryotic translation initiation factor 2α(e IF2α)-activating transcription factor 4(ATF4)-CCAAT/enhancer binding protein homologous protein(CHOP) signaling pathway by Pollen Typhae total flavone(PTF). Methods: Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group(70 μg/m L high ox-LDL), an HPTF group(70 μg/m L high ox-LDL+500 μg/m L PTF), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), and a LPTF group(70 μg/m L high ox-LDL+100 μg/m L PTF) in the first part;and a normal control group, an ox-LDL group(70 μg/mL high ox-LDL), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), a sh RNA group(transducted with PERK shRNA lentiviral particles), a scramble shRNA group(transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group(250 μg/mL PTF+70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group(70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their m RNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was applied to test cel viability, and the level of apoptosis was monitored by flow cytometry. Results: The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and sh RNA groups. Moreover, the ox-LDL group had increased protein and m RNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK, e IF2α, ATF4 and CHOP, which were attenuated in PTF treatment groups and sh RNA groups. Conclusions: The apoptosis induced by ox-LDL had a strong relation to ER stress. The protective effect of PTF on ER stressinduced apoptosis was associated with inhibition of the PERK-eIF2α-ATF4-CHOP pathway, which might be a potential therapeutic strategy for enhancing the stability of atherosclerotic plaques.
