桔梗皂苷D经CYP的代谢活化在抑制A549细胞活性中的作用研究  

Effect of platycodon D on inhibition of A549 cell activity by CYP metabolism activation

作　　者：宋丹妮 陈苏勤 管文燕[2] 周徐陈 张正光[3] 郭园园[3] SONG Danni;CHEN Suqin;GUAN Wenyan;ZHOU Xuchen;ZHANG Zhengguang;GUO Yuanyuan(School of Nursing,Nanjing Medical University,Nanjing 211100,Jiangsu,China;Nanjing Gulou Hospital,Nanjing 210008,Jiangsu,China;School of Medicine&Holistic Integrative Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China)

机构地区：[1]南京医科大学护理学院,江苏南京211100 [2]南京鼓楼医院,江苏南京210008 [3]南京中医药大学医学院·整合医学学院,江苏南京210023

出　　处：《现代中西医结合杂志》2022年第19期2647-2651,2662,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金青年项目(82004081,82004118);南京中医药大学自然科学基金项目(NZY82004081)。

摘　　要：目的 探究细胞色素P450酶(CYP450s)对桔梗皂苷D的代谢活化及对人非小细胞肺癌细胞株(A549细胞)的活性影响。方法 构建稳定表达CYP1A1、CYP1A2、CYP2B6、CYP2E1的人非小细胞肺癌A549细胞系,分别加入不同浓度(0,1,2,4μmol/L)的桔梗皂苷D孵育24 h后,采用CCK-8法检测各A549细胞系活性;将桔梗皂苷D与CYP1A1、CYP1A2、CYP2B6、CYP2E1重组代谢酶进行体外孵育120 min,超高效液相色谱-质谱(UPLC-MS)法检测药物浓度变化,并使用分子对接模型对桔梗皂苷D与CYP450s的结合位点和结合力进行预测分析。结果 成功构建稳定表达CYP1A1、CYP1A2、CYP2B6、CYP2E1的A549细胞系,Western blot及PCR均能检测出目的条带。随着桔梗皂苷D浓度的增加,与另外3组细胞系相比,高表达CYP1A1组A549细胞活性被明显抑制(P均<0.05);与另外3组代谢酶相比,桔梗皂苷D与CYP1A1体外孵育120 min后含量显著降低(P均<0.05);分子对接模型显示,桔梗皂苷D与CYP1A1有较强的结合力,氢键、盐桥及疏水相互作用为主要的相互作用力。结论 桔梗皂苷D能被CYP1A1代谢活化,并增强对肺癌细胞的活性抑制作用。Objective It is to explore the effect of cytochrome P450 enzyme(CYP450s)on the metabolic activation of platycodon D(PD)and the activity of human non-small cell lung cancer cell line(A549).Methods Human non-small cell lung cancer cell line(A549)with stable expression of CYP1A1,CYP1A2,CYP2B6 and CYP2E1 was constructed,and were respectively incubated for 24 hours with 0μmol/L PD,1μmol/L PD,2μmol/L PD,4μmol/L PD,and the activation of every A549 cell line were detected by CCK-8 method.PD was incubated with recombinant metabolic enzymes CYP1A1,CYP1A2,CYP2B6 and CYP2E1 for 120min in vitro.The change of drug concentration was detected by UPLC-MS,and the binding site and binding force of PD and CYP450 enzyme were predicted and analyzed by molecular docking model.Results A549 cell lines with stable expression of CYP1A1,CYP1A2,CYP2B6 and CYP2E1 were successfully constructed,and the target bands could be detected by Western blot and PCR.With the increase of PD concentration,the activity of A549 cells in CYP1A1 high expression group was inhibited compared with the other three groups(P<0.05);Compared with the other three groups,the contents of PD and CYP1A1 were significantly decreased after in vitro incubation for 120 min(P<0.05);Molecular docking model showed that PD and CYP1A1 had strong binding force,and hydrogen bond,salt bridge and hydrophobic interaction were the main interaction forces.Conclusion PD can be metabolized and activated by CYP1A1,and its inhibitory effect on lung cancer cells can be enhanced.

关 键 词：桔梗皂苷D 细胞色素P450酶 肺癌 A549细胞 

分 类 号：R-32[医药卫生]