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目的用分子对接技术探讨苍耳亭在上皮间质转化(EMT)过程中的作用靶点,并通过Western Blotting实验检测其对肝癌HepG2细胞相应靶点蛋白表达的影响。方法选取在EMT过程中的关键因子蓬乱蛋白(Dhs)、波形蛋白(Vimentin)、Snail、血管内皮生长因子受体3(VEGFR3)为作用靶点,采用分子对接虚拟技术评价苍耳亭与其空间结合能力,并与相应内源性物质烟酰胺腺嘌呤二核苷酸、醋酸离子、黄素腺嘌呤二核苷酸、N-乙酰葡糖胺对比;培养HepG2细胞,给予1、5、20μmol/L浓度的苍耳亭,利用Western Blotting实验检测其对Dhs、Vimentin、Snail、VEGFR3蛋白表达的影响。结果分子对接结果显示,苍耳亭与EMT过程中作用靶点均具有一定亲和力,其中与Dhs、Snail、VEGFR3的亲和力高于内源性物质,与Vimentin的亲和能力不及内源性物质;Western Blotting实验结果显示,苍耳亭显著下调Vimentin、Snail、VEGFR3蛋白的表达,显著上调E-cadherin蛋白表达(P<0.05、0.01、0.001)。结论苍耳亭对肝癌侵袭转移关键因子E-cadherin、Vimentin、Snail、VEGFR3有明显影响,可能是其潜在靶点;分子虚拟对接和Western blotting实验结果具有一定的相似性,能预先提示潜在靶因子。
Objective To explore the role of Xuanmiting in the process of epithelial-mesenchymal transition (EMT) by molecular docking and to detect the effect of Xanthium on the target protein of HepG2 cells by Western Blotting. Methods Dhs, Vimentin, Snail and vascular endothelial growth factor receptor 3 (VEGFR3) were selected as the target of EMT. The molecular docking virtual technique was used to evaluate the relationship between Xuan ear pavilion and its space Ability, and with the corresponding endogenous nicotinamide adenine dinucleotide, acetate ion, flavin adenine dinucleotide, N-acetylglucosamine contrast; cultured HepG2 cells were given 1,5,20 μmol / L Concentration of Xanthium Pavilion, using Western Blotting test to detect Dhs, Vimentin, Snail, VEGFR3 protein expression. The results of molecular docking showed that Xanthium Pavilion has some affinity with the target of EMT. The affinity with Dhs, Snail and VEGFR3 is higher than that of endogenous substances, and the affinity with Vimentin is less than that of endogenous substances. The results of Blotting showed that the expression of Vimentin, Snail and VEGFR3 protein was significantly decreased in Xuantuting group, and the expression of E-cadherin protein was significantly increased (P <0.05,0.01,0.001). Conclusion Xuan earmule has obvious influence on the key factors of invasion and metastasis of HCC, such as E-cadherin, Vimentin, Snail and VEGFR3, which may be potential target. The results of molecular virtual docking and Western blotting have some similarities, which can preliminarily suggest potential target factor .