目的:采用网络药理学方法探讨开心散治疗抑郁症和抑郁症合并心肌梗死(myocardial infarction,MI)的主要成分、作用靶点、传导通路和作用机制的异同并对关键靶点进行实验验证。方法:基于公共数据库结合ADME算法筛选开心散有效成分和作用...目的:采用网络药理学方法探讨开心散治疗抑郁症和抑郁症合并心肌梗死(myocardial infarction,MI)的主要成分、作用靶点、传导通路和作用机制的异同并对关键靶点进行实验验证。方法:基于公共数据库结合ADME算法筛选开心散有效成分和作用靶点;通过DisGeNET数据库筛选抑郁症和MI及其共病靶点;利用STRING 11.0数据库构建蛋白-蛋白互作(protein-protein interactions,PPI)网络;通过核心靶基因导入基因功能注释数据库(the database for annotation,visualization and integrated discovery,DAVID)进行京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)靶点相关通路分析;通过Cytoscape构建“药物成分-靶点-通路”网络;分析药物成分血脑屏障通过性,并采用Autodock Vina软件进行分子对接验证;建立抑郁症体外模型,采用蛋白质免疫印迹法(western blot,WB)对关键蛋白表达进行验证。结果:筛选出抑郁症与MI共同靶点28个;开心散主要成分30种,治疗抑郁症潜在靶点36个,其中关键靶点为Akt1、TNF、MAPK8、JUN、PTGS2、IL1B、IFNG、GSK3B、NOS2、DRD1等;开心散治疗抑郁症合并MI的主要成分14种,共同作用关键靶点为Akt1、TNF、IL1B、GSK3B、NOS2、BAX、NR3C2、NOS3、ESR1、ADRB1;开心散治疗抑郁症的成分较治疗抑郁症合并MI共同成分具有更强的血脑屏障通过性;分子对接结果显示山柰酚作用于Akt1靶点,参与调控PI3K/Akt通路;体外实验证实山柰酚可上调抑郁模型细胞Akt1蛋白表达。结论:开心散治疗抑郁症和抑郁症合并MI成分和靶点既有相同之处也有不同之处,体现“异病同治”的特点,且其治疗抑郁症主要侧重于对中枢神经系统和内分泌系统的调节,验证实验证实开心散对于治疗抑郁症和抑郁症合并MI具有有效性。展开更多
目的:探究抗AGGF1中和抗体(RDD-Ab),RDD-Ab + 抗CTLA-4抗体联合治疗对小鼠黑色素瘤的治疗作用。方法:合成AGGF1的GTFQRDDAPASVHSE肽并制备多克隆中和抗体(RDD-Ab)。为了评估RDD-Ab对血管生成活性和黑色素瘤生长的影响,我们实施了小管生...目的:探究抗AGGF1中和抗体(RDD-Ab),RDD-Ab + 抗CTLA-4抗体联合治疗对小鼠黑色素瘤的治疗作用。方法:合成AGGF1的GTFQRDDAPASVHSE肽并制备多克隆中和抗体(RDD-Ab)。为了评估RDD-Ab对血管生成活性和黑色素瘤生长的影响,我们实施了小管生成实验、迁移实验、细胞增殖实验、黑色素瘤细胞皮下移植性模型实验和免疫组化等实验。结果:实验结果显示,制备的RDD-Ab可以识别细胞中天然的AGGF1蛋白和过表达的AGGF1蛋白。RDD-Ab可显著抑制血管内皮细胞小管形成、迁移和增殖。与IgG对照组相比,RDD-Ab治疗显著减缓黑色素瘤生长,RDD-Ab + CTLA-4抗体联合治疗时肿瘤生长速度最慢。免疫组化实验也表明,RDD-Ab显著减少瘤内微血管生成与肿瘤细胞增殖,同时,联合治疗可显著增加瘤内CD4+和CD8+淋巴细胞的浸润。结论:RDD-Ab可在体外抑制血管内皮细胞的血管新生功能,具有高效的黑色素瘤生长抑制作用,RDD-Ab + CTLA-4联合治疗黑色素瘤效果更佳(显著增加瘤内CD4+和CD8+淋巴细胞的浸润),这为未来黑色素瘤的临床干预提供了一种新的潜在治疗方案。Objective: To investigate the therapeutic effect of neutralizing antibody (against AGGF1, RDD-Ab), RDD-Ab + CTLA-4 therapy on mouse melanoma. Methods: Synthesize GTFQRDAPASVHSE peptide of AGGF1 and prepare polyclonal neutralizing antibody (RDD-Ab). In order to evaluate the effects of RDD-Ab on angiogenesis and melanoma growth, we conducted tube formation, migration, cell proliferation, subcutaneous melanoma cell transplantation model and immunohistochemistry (IHC). Results: RDD-Ab can recognize both natural AGGF1 protein and overexpressed AGGF1 protein in cells. The experimental results showed that RDD-Ab significantly inhibits the formation, migration, and proliferation of endothelial cell tubules. Compared with the IgG control group, RDD-Ab significantly slowed down the growth of melanoma. IHC experiments showed that RDD-Ab significantly suppressed tumor angiogenesis and proliferation. The combination therapy of RDD-Ab + CTLA-4 antibody has the slowest tumor growth rate, and the combination therapy increases the infiltration of CD4+ and CD8+ lymphocytes robustly in solid tumors. Conclusion: RDD-Ab can inhibit the angiogenesis function of endothelial cells in vitro and has a highly effective inhibitory effect on melanoma growth has an efficient inhibitory effect on melanoma growth, and the combined treatment with CTLA-4 antibody is more effective. This provides a new potential treatment option for clinical intervention of melanoma in the future.展开更多
目的:探究AGGF1预处理的EPCs对肾IRI的治疗作用。方法:用淋巴细胞分离液(histopaque-1083)离心分离出单个核细胞层,分离的单个核细胞(Bone marrow mononuclear cells, MNCs)进行鉴定(Dil-ac-LDL和FITC-UEA-1染色)后用分离纯化的AGGF1蛋...目的:探究AGGF1预处理的EPCs对肾IRI的治疗作用。方法:用淋巴细胞分离液(histopaque-1083)离心分离出单个核细胞层,分离的单个核细胞(Bone marrow mononuclear cells, MNCs)进行鉴定(Dil-ac-LDL和FITC-UEA-1染色)后用分离纯化的AGGF1蛋白预处理(0.5 μg/mL,洗涤后尾静脉注射) 12 h,之后进行小鼠细胞治疗。小鼠分Sham组、生理盐水组(Saline组,I/R小鼠)、EPCs组(移植EPCs,I/R小鼠)、AGGF1-EPCs组(移植AGGF1蛋白预处理的EPCs,I/R小鼠)共4组,为了评估AGGF1预处理的EPCs植入疗法对肾IRI的影响,我们实施了免疫荧光实验、免疫组化和ELISA等实验。结果:实验结果显示,MNCs来源的EPCs呈现Dil-ac-LDL和FITC-UEA-1阳性。ELISA实验结果显示,AGGF1-EPCs组小鼠的尿素氮水平(BUN),肌酐(Creatine)与急性肾损伤标志物NGAL水平均较Saline组、EPCs组显著下降,同时,免疫组化分析也显示AGGF1-EPCs组小鼠肾脏的F4/80较Saline组、EPCs组显著下降。此外,血清IL-1β,TNF-α的检测结果也显示,炎症因子(IL-1β, TNF-α)在AGGF1-EPCs组小鼠的中表达最低。结论:AGGF1预处理的EPCs植入疗法可以显著降低缺血再灌注引起的急性肾损伤与炎症反应,对IRI具有良好的改善保护作用,这为未来肾IR的临床干预提供了一种新的潜在治疗方案。Objective: To explore the therapeutic effect of AGGF1 primed-EPCs on renal ischemia-reperfusion injury (IRI). Methods: We centrifuge and separate the bone marrow mononuclear cells (MNCs) using histopaque-1083, and identify the MNCs with Dil-ac-LDL and FITC-UEA-1 staining. After pretreatment with purified AGGF1 protein (0.5 μg/mL), EPCs were washed and injected into the C57BL/6J mice via tail vein. The mice were divided into four groups: Sham group, Saline group (I/R mice), EPCs group (transplanted EPCs, I/R mice), and AGGF1-EPCs group (EPCs pretreated with AGGF1 protein, I/R mice). To evaluate the effect of EPCs implantation on renal IRI, we carried out western blot, ELISA, and immunohistochemistry experiments. Results: Our results showed that EPCs derived from MNCs exhibited Dil-ac-LDL and FITC-UEA-1 positive. The ELISA results showed that the levels of urea nitrogen (BUN), creatinine and acute kidney injury marker NGAL in the AGGF1-EPCs group mice were significantly lower than those in the Saline group and EPCs group. At the same time, immunohistochemical analysis also showed that the NGAL levels and F4/80 in the kidneys of AGGF1-EPCs group mice were significantly lower than those in the Saline group and EPCs group. In addition, we also observed that the inflammatory factors IL-1β, TNF-α were significantly reduced in the AGGF1-EPCs group of mice. Conclusion: The implantation therapy of AGGF1 primed-EPCs significantly reduce acute renal injury and inflammatory response caused by ischemia-reperfusion. This provides a new potential treatment for clinical intervention of renal IR.展开更多
文摘目的:采用网络药理学方法探讨开心散治疗抑郁症和抑郁症合并心肌梗死(myocardial infarction,MI)的主要成分、作用靶点、传导通路和作用机制的异同并对关键靶点进行实验验证。方法:基于公共数据库结合ADME算法筛选开心散有效成分和作用靶点;通过DisGeNET数据库筛选抑郁症和MI及其共病靶点;利用STRING 11.0数据库构建蛋白-蛋白互作(protein-protein interactions,PPI)网络;通过核心靶基因导入基因功能注释数据库(the database for annotation,visualization and integrated discovery,DAVID)进行京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)靶点相关通路分析;通过Cytoscape构建“药物成分-靶点-通路”网络;分析药物成分血脑屏障通过性,并采用Autodock Vina软件进行分子对接验证;建立抑郁症体外模型,采用蛋白质免疫印迹法(western blot,WB)对关键蛋白表达进行验证。结果:筛选出抑郁症与MI共同靶点28个;开心散主要成分30种,治疗抑郁症潜在靶点36个,其中关键靶点为Akt1、TNF、MAPK8、JUN、PTGS2、IL1B、IFNG、GSK3B、NOS2、DRD1等;开心散治疗抑郁症合并MI的主要成分14种,共同作用关键靶点为Akt1、TNF、IL1B、GSK3B、NOS2、BAX、NR3C2、NOS3、ESR1、ADRB1;开心散治疗抑郁症的成分较治疗抑郁症合并MI共同成分具有更强的血脑屏障通过性;分子对接结果显示山柰酚作用于Akt1靶点,参与调控PI3K/Akt通路;体外实验证实山柰酚可上调抑郁模型细胞Akt1蛋白表达。结论:开心散治疗抑郁症和抑郁症合并MI成分和靶点既有相同之处也有不同之处,体现“异病同治”的特点,且其治疗抑郁症主要侧重于对中枢神经系统和内分泌系统的调节,验证实验证实开心散对于治疗抑郁症和抑郁症合并MI具有有效性。
文摘目的:探究抗AGGF1中和抗体(RDD-Ab),RDD-Ab + 抗CTLA-4抗体联合治疗对小鼠黑色素瘤的治疗作用。方法:合成AGGF1的GTFQRDDAPASVHSE肽并制备多克隆中和抗体(RDD-Ab)。为了评估RDD-Ab对血管生成活性和黑色素瘤生长的影响,我们实施了小管生成实验、迁移实验、细胞增殖实验、黑色素瘤细胞皮下移植性模型实验和免疫组化等实验。结果:实验结果显示,制备的RDD-Ab可以识别细胞中天然的AGGF1蛋白和过表达的AGGF1蛋白。RDD-Ab可显著抑制血管内皮细胞小管形成、迁移和增殖。与IgG对照组相比,RDD-Ab治疗显著减缓黑色素瘤生长,RDD-Ab + CTLA-4抗体联合治疗时肿瘤生长速度最慢。免疫组化实验也表明,RDD-Ab显著减少瘤内微血管生成与肿瘤细胞增殖,同时,联合治疗可显著增加瘤内CD4+和CD8+淋巴细胞的浸润。结论:RDD-Ab可在体外抑制血管内皮细胞的血管新生功能,具有高效的黑色素瘤生长抑制作用,RDD-Ab + CTLA-4联合治疗黑色素瘤效果更佳(显著增加瘤内CD4+和CD8+淋巴细胞的浸润),这为未来黑色素瘤的临床干预提供了一种新的潜在治疗方案。Objective: To investigate the therapeutic effect of neutralizing antibody (against AGGF1, RDD-Ab), RDD-Ab + CTLA-4 therapy on mouse melanoma. Methods: Synthesize GTFQRDAPASVHSE peptide of AGGF1 and prepare polyclonal neutralizing antibody (RDD-Ab). In order to evaluate the effects of RDD-Ab on angiogenesis and melanoma growth, we conducted tube formation, migration, cell proliferation, subcutaneous melanoma cell transplantation model and immunohistochemistry (IHC). Results: RDD-Ab can recognize both natural AGGF1 protein and overexpressed AGGF1 protein in cells. The experimental results showed that RDD-Ab significantly inhibits the formation, migration, and proliferation of endothelial cell tubules. Compared with the IgG control group, RDD-Ab significantly slowed down the growth of melanoma. IHC experiments showed that RDD-Ab significantly suppressed tumor angiogenesis and proliferation. The combination therapy of RDD-Ab + CTLA-4 antibody has the slowest tumor growth rate, and the combination therapy increases the infiltration of CD4+ and CD8+ lymphocytes robustly in solid tumors. Conclusion: RDD-Ab can inhibit the angiogenesis function of endothelial cells in vitro and has a highly effective inhibitory effect on melanoma growth has an efficient inhibitory effect on melanoma growth, and the combined treatment with CTLA-4 antibody is more effective. This provides a new potential treatment option for clinical intervention of melanoma in the future.
文摘目的:探究AGGF1预处理的EPCs对肾IRI的治疗作用。方法:用淋巴细胞分离液(histopaque-1083)离心分离出单个核细胞层,分离的单个核细胞(Bone marrow mononuclear cells, MNCs)进行鉴定(Dil-ac-LDL和FITC-UEA-1染色)后用分离纯化的AGGF1蛋白预处理(0.5 μg/mL,洗涤后尾静脉注射) 12 h,之后进行小鼠细胞治疗。小鼠分Sham组、生理盐水组(Saline组,I/R小鼠)、EPCs组(移植EPCs,I/R小鼠)、AGGF1-EPCs组(移植AGGF1蛋白预处理的EPCs,I/R小鼠)共4组,为了评估AGGF1预处理的EPCs植入疗法对肾IRI的影响,我们实施了免疫荧光实验、免疫组化和ELISA等实验。结果:实验结果显示,MNCs来源的EPCs呈现Dil-ac-LDL和FITC-UEA-1阳性。ELISA实验结果显示,AGGF1-EPCs组小鼠的尿素氮水平(BUN),肌酐(Creatine)与急性肾损伤标志物NGAL水平均较Saline组、EPCs组显著下降,同时,免疫组化分析也显示AGGF1-EPCs组小鼠肾脏的F4/80较Saline组、EPCs组显著下降。此外,血清IL-1β,TNF-α的检测结果也显示,炎症因子(IL-1β, TNF-α)在AGGF1-EPCs组小鼠的中表达最低。结论:AGGF1预处理的EPCs植入疗法可以显著降低缺血再灌注引起的急性肾损伤与炎症反应,对IRI具有良好的改善保护作用,这为未来肾IR的临床干预提供了一种新的潜在治疗方案。Objective: To explore the therapeutic effect of AGGF1 primed-EPCs on renal ischemia-reperfusion injury (IRI). Methods: We centrifuge and separate the bone marrow mononuclear cells (MNCs) using histopaque-1083, and identify the MNCs with Dil-ac-LDL and FITC-UEA-1 staining. After pretreatment with purified AGGF1 protein (0.5 μg/mL), EPCs were washed and injected into the C57BL/6J mice via tail vein. The mice were divided into four groups: Sham group, Saline group (I/R mice), EPCs group (transplanted EPCs, I/R mice), and AGGF1-EPCs group (EPCs pretreated with AGGF1 protein, I/R mice). To evaluate the effect of EPCs implantation on renal IRI, we carried out western blot, ELISA, and immunohistochemistry experiments. Results: Our results showed that EPCs derived from MNCs exhibited Dil-ac-LDL and FITC-UEA-1 positive. The ELISA results showed that the levels of urea nitrogen (BUN), creatinine and acute kidney injury marker NGAL in the AGGF1-EPCs group mice were significantly lower than those in the Saline group and EPCs group. At the same time, immunohistochemical analysis also showed that the NGAL levels and F4/80 in the kidneys of AGGF1-EPCs group mice were significantly lower than those in the Saline group and EPCs group. In addition, we also observed that the inflammatory factors IL-1β, TNF-α were significantly reduced in the AGGF1-EPCs group of mice. Conclusion: The implantation therapy of AGGF1 primed-EPCs significantly reduce acute renal injury and inflammatory response caused by ischemia-reperfusion. This provides a new potential treatment for clinical intervention of renal IR.
