异类叶升麻苷和桔梗皂苷D的体外抗肺癌研究

Study on the inhibitory effect of the isoverbascoside and platycodin D on anti-lung cancer in vitro

目的:探讨异类叶升麻苷与桔梗皂苷D单用及配伍对A549细胞增殖、迁移和侵袭的影响及作用机制。方法:采用CCK-8、细胞划痕、Transwell试验分别检测细胞增殖、迁移和侵袭,Western Blot检测EphA2/AKT/mTOR通路蛋白,ELISA检测肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)含量。结果:异类叶升麻苷和桔梗皂苷D均能抑制细胞增殖,且IC50分别为141、170μg/mL。与对照组比较,桔梗组、配伍组愈合率显著下降(P<0.01),给药各组穿膜细胞数均显著减少(P<0.01),其中配伍组最为明显。与对照组比较,桔梗组、配伍组显著下调EphA2、p-EphA2蛋白表达(P<0.01),桔梗组显著下调p-AKT、mTOR蛋白表达(P<0.01)。与对照组比较,桔梗组IL-6、TNF-α显著升高(P<0.01,P<0.05),而异类叶组TNF-α和配伍组IL-6显著下调(P<0.01)。结论:异类叶升麻苷、桔梗皂苷D通过调控EphA2/AKT/mTOR通路和调节TNF-α、IL-6抑制A549细胞增殖、迁移和侵袭。且二者配伍显著调控Eph A2因子,并能减轻桔梗诱导IL-6和TNF-α的上调,推测配伍组可能具有增效减毒的作用。

Objective:To explore the effects and mechanism of the two components of isoverbascoside and platycodin D(PD)on the proliferation,migration and invasion of A549 cells.Methods:Proliferation,migration and invasion of cells were detected by CCK-8,cell scratch and Transwell assay,EphA2/AKT/mTOR pathway protein expression was detected by Western Blot,and TNF-αand IL-6 content was detected by ELISA.Results:Isoverbascoside and PD could inhibit cell proliferation with respectively IC50 of 141 and 170μg/mL.Compared with the control group,the healing rate of the PD group and the mixed group decreased significantly(P<0.01),the number of transmembrane cells in each group was significantly decreased(P<0.01),and the mixed group was the most significant.Compared with the control group,the EphA2 and p-EphA2 proteins were significantly down-regulated in PD group and mixed group(P<0.01),p-Akt and mTOR protein was significantly down-regulated in PD group(P<0.01).Compared with the control group,TNF-αand IL-6 increased in PD group(P<0.01,P<0.05),while TNF-αin the isoverbascoside group and IL-6 in the mixed group were significantly down-regulated(P<0.01).Conclusion:Isoacteoside and PD can inhibit the migration and invasion of A549 cells by regulating EphA2/AKT/mTOR pathway and regulating TNF-αand IL-6.In addition,the mixed group significantly regulates EphA2 factor,and can reduce the increase of IL-6 and TNF-αof PD.It is speculated that the mixed group may have the effect of increasing efficiency and reducing toxicity.