基于分子对接的合欢皂苷J_(8)抑制血管内皮细胞增殖作用靶点及其凋亡相关细胞信号通路研究

Study on the target of julibroside J_(8) inhibiting the proliferation of vascular endothelial cells and its apoptosis related cell signal pathways based on molecular docking technology

为探讨合欢皂苷J_(8)(julibroside J_(8),J8)抑制肿瘤血管内皮细胞增殖作用靶点及其相关细胞凋亡的信号通路。本文采用HPLC法检测内皮细胞经J8作用后,J8的在细胞内外含量变化;Vina软件将J8与VEGF、FAS、DR3、DR4、DR5、TFR-1进行分子对接分析;Western blot法检测在加药前后,内皮细胞中VEGF、p-JNK、Bax、EnDOG、Caspase-3、Caspase-8以及Caspase-9蛋白表达水平的变化。分子对接结果表明VEGF、FAS对应的靶点蛋白与J8结合性能较好且多位点结合。HUVEC细胞在加入J8作用24 h后,VEGF、p-JNK等蛋白表达明显下调,并可显著上调诱导凋亡相关蛋白Bax和EnDOG的表达,而且对Caspase-3、Caspase-8以及Caspase-9的表达水平无显著的影响。J8可能是通过与血管内皮细胞膜表面的VEGF结合,抑制血管内皮细胞增殖,通过削弱VEGF/JNK通路活性从而引起内皮细胞凋亡。

To investigate the target of julibroside J_(8)(J8)inhibiting the proliferation of tumor vascular endothelial cells and its related signal pathway of apoptosis.In this paper,HPLC method was used to detect the content change of J8 in and out of endothelial cells after being treated with J8;Vina software docked J8 with VEGF,FAS,DR3,DR4,DR5,TFR-1 for analysis;Western blot method was used to detect the changes in the expression levels of VEGF,p-JNK,Bax,ENDOG,Caspase-3,Caspase-8,and Caspase-9 proteins in endothelial cells before and after drug addition.The results of molecular docking showed that the target proteins corresponding to VEGF and FAS had good binding ability to J8 and could bind to multiple sites.After HUVEC cells were treated with J8 for 24 h,the expressions of VEGF,p-JNK and other proteins were significantly decreased,and the expressions of apoptosis inducing proteins Bax and EnDOG were significantly up-regulated,and there was no significant effect on the expression levels of Caspase-3,Caspase-8 and Caspase-9.J8 may inhibit the proliferation of vascular endothelial cells by binding with VEGF on the surface of vascular endothelial cell membrane,and cause endothelial cell apoptosis by weakening the activity of VEGF/JNK pathway.