Objective: To clarify the inhibitory mechanism of Haizao Yuhu Decoction on BRAFV600E mutation-driven papillary thyroid carcinoma cells. Methods: Prepare seaweed Yuhu Decoction medicated serum, select human normal thyr...Objective: To clarify the inhibitory mechanism of Haizao Yuhu Decoction on BRAFV600E mutation-driven papillary thyroid carcinoma cells. Methods: Prepare seaweed Yuhu Decoction medicated serum, select human normal thyroid cells Nthy-ori3-1 as the normal control group, and PLX4032 as the positive control drug. The experiment was divided into normal control group, model control group, PLX4032 group, Haizao Yuhu Decoction group (referred to as HYD group), Haizao Yuhu Decoction group + PLX4032 (referred to as HYD+PLX4032 group), and high iodine water group, at 8h and 24h respectively. At 72h, the cell proliferation of each group was detected by MTT method;at 24h, the expression of ERK and p-ERK protein in each group was detected by Western blot. Results: The longer the time, the more obvious the inhibitory effect of Haizao Yuhu Decoction-containing serum on the proliferation activity of BCPAP cells: Compared with the normal control group, the BCPAP cell proliferation activity of the model control group was significantly enhanced at 24h and 72h (P <0.05);At 24h, the PLX4032 group, HYD group, and HYD+PLX4032 group all showed a tendency to inhibit the proliferation of BCPAP cells, but there was no statistical difference;at 72h, compared with the model control group, the PLX4032 group The cell proliferation activity of the HYD group and HYD+PLX4032 group was significantly inhibited (P <0.05), and there was no difference between the high iodine water group and the model control group (P>0.05). In inhibiting the proliferation of BCPAP cells, the medicated serum of Shanghai Zaoyuhu Decoction has a synergistic effect with PLX4032 (F=10.87, P=0.005). Western blot results showed that there was no difference in the expression of ERK1/2 protein between the groups, but there were significant differences in the expression of p-ERK1/2 protein between the groups: Compared with the normal control group, the expression of p-ERK1/2 protein in the model group increased significantly (P < 0.05);Compared with the model control group, the expression of p-ERK1/2 protein in the PLX4032 group, the HYD group, and the HYD+PLX4032 group decreased significantly (P <0.05), and there was no difference between the high iodine water group and the model control group (P> 0.05). Conclusion: The medicated serum of Haizao Yuhu Decoction has an inhibitory effect on the proliferation of BCPAP cells, and its mechanism may be inhibiting the proliferation of BCPAP cells by inhibiting ERK1/2 protein phosphorylation, a protein post-translational modification process;the medicated serum of Haizao Yuhu Decoction may have Help enhance the effect of PLX4032 curative effect.展开更多
目的:基于网络药理学和分子对接技术探究海藻、昆布治疗Graves病的作用机制。方法:借助中药系统药理学数据库与分析平台(TCMSP)获得中药海藻、昆布的活性成分、靶点,运用蛋白质数据库Uniprot将靶点的基因名称规范化。分别在GeneCards、O...目的:基于网络药理学和分子对接技术探究海藻、昆布治疗Graves病的作用机制。方法:借助中药系统药理学数据库与分析平台(TCMSP)获得中药海藻、昆布的活性成分、靶点,运用蛋白质数据库Uniprot将靶点的基因名称规范化。分别在GeneCards、OMIM、DrugBank等数据库上检索Graves疾病靶点,使用R语言绘制海藻、昆布与Graves病交集靶点的韦恩图,并通过Cytoscape 3.8.2软件绘制“海藻、昆布-Graves病”活性成分-靶点图以及蛋白质-蛋白质相互作用网络图。利用DAVID数据库对药物与疾病交集基因进行基因本体论(GO)功能分析、京都基因与基因组百科全书(KEGG)通路富集分析。在PubChem数据库中下载小分子配体结构,在PDB数据库中下载蛋白受体结构,运用Autodock vina进行分子对接。结果:筛选出海藻、昆布与Graves病相关的活性成分11个、蛋白123个。GO功能富集分析显示,有生物过程585个、细胞组分63个、分子功能125个。KEGG通路富集分析显示,有相关通路149个。分子对接展现了具有较好相互作用的核心成分与核心蛋白的对接效果,核心成分有槲皮素和花生四烯酸,核心蛋白有Jun原癌基因(Jun Proto-Oncogene,JUN)、丝裂原活化蛋白激酶(MitogenActivated Protein Kinase,MAPK)1、肿瘤蛋白p53(Tumor Protein p53,TP53)。海藻、昆布可能通过调节晚期糖基化终末产物-晚期糖基化终末产物受体(AGE-RAGE)、肿瘤坏死因子(Tumor Necrosis Factor,TNF)等信号通路发挥治疗Graves病的作用。结论:海藻、昆布有效成分可能是槲皮素、花生四烯酸,它们分别通过JUN、MAPK1、TP53靶点蛋白,以及TNF-α、AGERAGE通路治疗Graves病。展开更多
基金Fund Project:General Program of Natural Science Foundation of Liaoning Province(No.2015020390)General Program of National Natural Science Foundation of China(No.81874441)。
文摘Objective: To clarify the inhibitory mechanism of Haizao Yuhu Decoction on BRAFV600E mutation-driven papillary thyroid carcinoma cells. Methods: Prepare seaweed Yuhu Decoction medicated serum, select human normal thyroid cells Nthy-ori3-1 as the normal control group, and PLX4032 as the positive control drug. The experiment was divided into normal control group, model control group, PLX4032 group, Haizao Yuhu Decoction group (referred to as HYD group), Haizao Yuhu Decoction group + PLX4032 (referred to as HYD+PLX4032 group), and high iodine water group, at 8h and 24h respectively. At 72h, the cell proliferation of each group was detected by MTT method;at 24h, the expression of ERK and p-ERK protein in each group was detected by Western blot. Results: The longer the time, the more obvious the inhibitory effect of Haizao Yuhu Decoction-containing serum on the proliferation activity of BCPAP cells: Compared with the normal control group, the BCPAP cell proliferation activity of the model control group was significantly enhanced at 24h and 72h (P <0.05);At 24h, the PLX4032 group, HYD group, and HYD+PLX4032 group all showed a tendency to inhibit the proliferation of BCPAP cells, but there was no statistical difference;at 72h, compared with the model control group, the PLX4032 group The cell proliferation activity of the HYD group and HYD+PLX4032 group was significantly inhibited (P <0.05), and there was no difference between the high iodine water group and the model control group (P>0.05). In inhibiting the proliferation of BCPAP cells, the medicated serum of Shanghai Zaoyuhu Decoction has a synergistic effect with PLX4032 (F=10.87, P=0.005). Western blot results showed that there was no difference in the expression of ERK1/2 protein between the groups, but there were significant differences in the expression of p-ERK1/2 protein between the groups: Compared with the normal control group, the expression of p-ERK1/2 protein in the model group increased significantly (P < 0.05);Compared with the model control group, the expression of p-ERK1/2 protein in the PLX4032 group, the HYD group, and the HYD+PLX4032 group decreased significantly (P <0.05), and there was no difference between the high iodine water group and the model control group (P> 0.05). Conclusion: The medicated serum of Haizao Yuhu Decoction has an inhibitory effect on the proliferation of BCPAP cells, and its mechanism may be inhibiting the proliferation of BCPAP cells by inhibiting ERK1/2 protein phosphorylation, a protein post-translational modification process;the medicated serum of Haizao Yuhu Decoction may have Help enhance the effect of PLX4032 curative effect.
文摘目的:基于网络药理学和分子对接技术探究海藻、昆布治疗Graves病的作用机制。方法:借助中药系统药理学数据库与分析平台(TCMSP)获得中药海藻、昆布的活性成分、靶点,运用蛋白质数据库Uniprot将靶点的基因名称规范化。分别在GeneCards、OMIM、DrugBank等数据库上检索Graves疾病靶点,使用R语言绘制海藻、昆布与Graves病交集靶点的韦恩图,并通过Cytoscape 3.8.2软件绘制“海藻、昆布-Graves病”活性成分-靶点图以及蛋白质-蛋白质相互作用网络图。利用DAVID数据库对药物与疾病交集基因进行基因本体论(GO)功能分析、京都基因与基因组百科全书(KEGG)通路富集分析。在PubChem数据库中下载小分子配体结构,在PDB数据库中下载蛋白受体结构,运用Autodock vina进行分子对接。结果:筛选出海藻、昆布与Graves病相关的活性成分11个、蛋白123个。GO功能富集分析显示,有生物过程585个、细胞组分63个、分子功能125个。KEGG通路富集分析显示,有相关通路149个。分子对接展现了具有较好相互作用的核心成分与核心蛋白的对接效果,核心成分有槲皮素和花生四烯酸,核心蛋白有Jun原癌基因(Jun Proto-Oncogene,JUN)、丝裂原活化蛋白激酶(MitogenActivated Protein Kinase,MAPK)1、肿瘤蛋白p53(Tumor Protein p53,TP53)。海藻、昆布可能通过调节晚期糖基化终末产物-晚期糖基化终末产物受体(AGE-RAGE)、肿瘤坏死因子(Tumor Necrosis Factor,TNF)等信号通路发挥治疗Graves病的作用。结论:海藻、昆布有效成分可能是槲皮素、花生四烯酸,它们分别通过JUN、MAPK1、TP53靶点蛋白,以及TNF-α、AGERAGE通路治疗Graves病。
