Background:Chronic pain is defined as pain that lasts for three months or even more.Over 25%of the global population suffers from chronic pain.Tong-luo Qu-tong(TLQT)Plaster is clinically used to treat arthritis.Howeve...Background:Chronic pain is defined as pain that lasts for three months or even more.Over 25%of the global population suffers from chronic pain.Tong-luo Qu-tong(TLQT)Plaster is clinically used to treat arthritis.However,its ability to treat chronic pain remains largely unknown.Methods:In this study,we explored the molecular mechanism of TLQT plaster in relieving chronic muscle pain by combining network pharmacology and RNA-seq analysis.We also applied the Elisa and the RT-qPCR.Results:We found 447 targets in TLQT and 13,599 targets related to chronic pain disease.And 419 intersecting targets were obtained,which mainly enrich the IL-17 signaling pathway,TNF signaling pathway,and Th17 cell differentiation pathway.Further,we constructed the SD rat model of chronic pain.The results of Von Frey Hair Test showed that the relief of muscle pain TLQT treated group was twice as much as that in the model group.The hot plate test results showed that the time of lifting the foot was 1.3 times as much as that of the model group.Moreover,TLQT effectively reduced the inflammation in rat muscle.With RNA-seq analysis,230 differentially expressed genes were collected.The RT-qPCR results indicated that the mRNA expression level of NCF1,CXCL10,and ICAM1 all promoted in the model group,and then decreased significantly in the TLQT treated group.The ELISA results performed that the level of IL-1βand IL-6 in TLQT group high dose group was reduced by about 1.6-fold,and for TNF-α,it was reduced by about 2.6-fold compared with the model group.Immunohistochemistry assay showed that the expression level of CXCL10 and ICAM1 was both up-regulated in the model group and down-regulated in the TLQT group.Conclusion:TLQT plaster reduces chronic muscle pain by inhibiting the expression of NCF1,CXCL10,and ICAM1 and reducing the level of muscle tissue inflammation.展开更多
目的采用网络药理学与分子对接技术探讨生脉注射液的活性成分和治疗新型冠状病毒肺炎(COVID-19)的潜在作用机制。方法利用TCMSP及BATMAN-TCM数据库筛选生脉注射液的活性化合物,通过TCMSP及Targetnet在线数据库预测作用靶点,通过Cytoscap...目的采用网络药理学与分子对接技术探讨生脉注射液的活性成分和治疗新型冠状病毒肺炎(COVID-19)的潜在作用机制。方法利用TCMSP及BATMAN-TCM数据库筛选生脉注射液的活性化合物,通过TCMSP及Targetnet在线数据库预测作用靶点,通过Cytoscape3.7.1构建活性成分-作用靶点网络图;在GeneCards及OMIM数据库中以"coronavirus pneumonia"为关键词搜索冠状病毒肺炎相关疾病靶点,与生脉注射液化合物靶点进行交集筛选出共同靶点作为研究靶点,将共同靶点导入STRING数据库获取数据后在Cytoscape 3.7.1软件中构建蛋白质-蛋白质相互作用网络图;利用R语言进行GO(gene ontology)功能、KEGG(Kyoto encyclopedia of genes and genomes)通路富集分析,预测其作用机制,并构建"成分-靶点-通路"网络图;通过DiscoveryStudio 2.5软件对关键靶点进行分子对接分析。结果生脉注射液筛选得到22个活性化合物,分别为邻苯二甲酸二辛酯、β-谷甾醇、当归酰基戈米辛O、戈米辛A、戈米辛R、五味子丙素、内南五味子酯乙、长南酸、南五味子内酯、香蒲木脂素B、新杜松烷酸A、新杜松烷酸B、新杜松烷酸C、新南五味子木脂宁、五味子内酯A、五味子内酯E、五味子酸、尿苷、薯蓣皂苷元、鸟嘌呤核苷、N-反式阿魏酰酪胺、豆甾醇。相应作用靶点224个,与COVID-19的共同靶点16个,分别为CASP3、CASP8、PTGS2、BCL2、BAX、PRKCA、PTGS1、PIK3CG、F10、NOS3、DPP4、NOS2、TLR9、ACE、ICAM1、PRKCE,关键靶点涉及CASP3、PTGS2、NOS2、NOS3、ICAM1。GO功能富集分析得到生物过程(BP)条目771个,细胞组成(CC)条目11个,分子功能(MF)条目79个。KEGG通路富集分析筛选得到67条(P<0.05)信号通路,主要涉及糖尿病并发症AGE-RAGE信号通路、凋亡通路、P53信号通路、小细胞肺癌通路等。分子对接结果显示与关键靶点对接较好的成分有五味子内酯E、豆甾醇、N-反式阿魏酰酪胺。结论生脉注射液中的活性化合物五味子内酯E、豆甾醇、N-反式阿魏酰酪胺等能作用于CASP3、PTGS2、NOS2、NOS3等靶点调节多条信号通路发挥抗炎、免疫调节、抗休克、增加血氧饱和度等作用,从而可能发挥对COVID-19的治疗作用。展开更多
文摘Background:Chronic pain is defined as pain that lasts for three months or even more.Over 25%of the global population suffers from chronic pain.Tong-luo Qu-tong(TLQT)Plaster is clinically used to treat arthritis.However,its ability to treat chronic pain remains largely unknown.Methods:In this study,we explored the molecular mechanism of TLQT plaster in relieving chronic muscle pain by combining network pharmacology and RNA-seq analysis.We also applied the Elisa and the RT-qPCR.Results:We found 447 targets in TLQT and 13,599 targets related to chronic pain disease.And 419 intersecting targets were obtained,which mainly enrich the IL-17 signaling pathway,TNF signaling pathway,and Th17 cell differentiation pathway.Further,we constructed the SD rat model of chronic pain.The results of Von Frey Hair Test showed that the relief of muscle pain TLQT treated group was twice as much as that in the model group.The hot plate test results showed that the time of lifting the foot was 1.3 times as much as that of the model group.Moreover,TLQT effectively reduced the inflammation in rat muscle.With RNA-seq analysis,230 differentially expressed genes were collected.The RT-qPCR results indicated that the mRNA expression level of NCF1,CXCL10,and ICAM1 all promoted in the model group,and then decreased significantly in the TLQT treated group.The ELISA results performed that the level of IL-1βand IL-6 in TLQT group high dose group was reduced by about 1.6-fold,and for TNF-α,it was reduced by about 2.6-fold compared with the model group.Immunohistochemistry assay showed that the expression level of CXCL10 and ICAM1 was both up-regulated in the model group and down-regulated in the TLQT group.Conclusion:TLQT plaster reduces chronic muscle pain by inhibiting the expression of NCF1,CXCL10,and ICAM1 and reducing the level of muscle tissue inflammation.
文摘目的采用短小棒状杆菌制剂对小白鼠ICAM1/LFA1基因表达的研究,从而进一步研究短小棒状杆菌制剂对免疫调节和抗肿瘤机理深入研究提出依据。方法本试验采用realtime PCR方法,以GAPDH和-βactin分别作为各组织样品的内源控制基因。结果短小棒状杆菌制剂短小棒状杆菌制剂小白鼠腹腔注射之后,ICAM1基因在脾脏组织中上调,5 h出现高峰,短小棒状杆菌制剂小白鼠腹腔注射之后3-5 h LFA-1基因持续性上调。不同组织的LAF-1基因检测结果显示,小肠和脾脏均表现为高表达,尤以脾脏突出。结论短小棒状杆菌制剂的免疫机理和治疗肿瘤的作用与早期激动免疫细胞的第一信号有直接相关。
文摘目的采用网络药理学与分子对接技术探讨生脉注射液的活性成分和治疗新型冠状病毒肺炎(COVID-19)的潜在作用机制。方法利用TCMSP及BATMAN-TCM数据库筛选生脉注射液的活性化合物,通过TCMSP及Targetnet在线数据库预测作用靶点,通过Cytoscape3.7.1构建活性成分-作用靶点网络图;在GeneCards及OMIM数据库中以"coronavirus pneumonia"为关键词搜索冠状病毒肺炎相关疾病靶点,与生脉注射液化合物靶点进行交集筛选出共同靶点作为研究靶点,将共同靶点导入STRING数据库获取数据后在Cytoscape 3.7.1软件中构建蛋白质-蛋白质相互作用网络图;利用R语言进行GO(gene ontology)功能、KEGG(Kyoto encyclopedia of genes and genomes)通路富集分析,预测其作用机制,并构建"成分-靶点-通路"网络图;通过DiscoveryStudio 2.5软件对关键靶点进行分子对接分析。结果生脉注射液筛选得到22个活性化合物,分别为邻苯二甲酸二辛酯、β-谷甾醇、当归酰基戈米辛O、戈米辛A、戈米辛R、五味子丙素、内南五味子酯乙、长南酸、南五味子内酯、香蒲木脂素B、新杜松烷酸A、新杜松烷酸B、新杜松烷酸C、新南五味子木脂宁、五味子内酯A、五味子内酯E、五味子酸、尿苷、薯蓣皂苷元、鸟嘌呤核苷、N-反式阿魏酰酪胺、豆甾醇。相应作用靶点224个,与COVID-19的共同靶点16个,分别为CASP3、CASP8、PTGS2、BCL2、BAX、PRKCA、PTGS1、PIK3CG、F10、NOS3、DPP4、NOS2、TLR9、ACE、ICAM1、PRKCE,关键靶点涉及CASP3、PTGS2、NOS2、NOS3、ICAM1。GO功能富集分析得到生物过程(BP)条目771个,细胞组成(CC)条目11个,分子功能(MF)条目79个。KEGG通路富集分析筛选得到67条(P<0.05)信号通路,主要涉及糖尿病并发症AGE-RAGE信号通路、凋亡通路、P53信号通路、小细胞肺癌通路等。分子对接结果显示与关键靶点对接较好的成分有五味子内酯E、豆甾醇、N-反式阿魏酰酪胺。结论生脉注射液中的活性化合物五味子内酯E、豆甾醇、N-反式阿魏酰酪胺等能作用于CASP3、PTGS2、NOS2、NOS3等靶点调节多条信号通路发挥抗炎、免疫调节、抗休克、增加血氧饱和度等作用,从而可能发挥对COVID-19的治疗作用。
