基于P糖蛋白研究仙茅对顺铂耐药非小细胞肺癌细胞的增敏机制  

Sensitivity-enhancing Mechanism of Curculiginis Rhizoma in Cisplatin-resistant Non-small Cell Lung Cancer Cells Based on P-glycoprotein

作　　者：郝雨 朱宝琛 薛春苗[1,2] 华国栋 高若瑜 黄鑫 刘文卉 王萌 HAO Yu;ZHU Baochen;XUE Chunmiao;HUA Guodong;GAO Ruoyu;HUANG Xin;LIU Wenhui;WANG Meng(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]北京中医药大学东直门医院,北京100700

出　　处：《世界中医药》2022年第24期3462-3466,3471,共6页World Chinese Medicine

基　　金：国家自然科学基金青年科学基金项目(82004125);北京中医药大学东直门医院2020科技创新专项(DZMKJCX-2020-004)。

摘　　要：目的:探究中药仙茅水煎液及其主要入血成分苔黑酚葡萄糖苷(OG)对非小细胞肺癌(NSCLC)顺铂耐药细胞(A549/cis)耐药性的影响及其潜在机制。方法:用AutoDock Vina分子对接软件评估P糖蛋白(P-gp)与OG的结合能力;CCK-8法检测顺铂、仙茅水煎液、OG及其联用对A549/cis细胞活力的抑制率;蛋白质免疫印迹法检测A549/cis中P-gp的表达水平。结果:P-gp与OG模拟分子对接的结合自由能为-7.6 kcal/mol(1 cal=4.184 J),二者具有良好的结合活性;顺铂对A549/cis的半抑制浓度(IC50)约为5.5μg/mL,仙茅水煎液与OG在梯度浓度下对A549/cis活力均无明显影响;顺铂低剂量(1.5μg/mL)与仙茅水煎液高、低剂量(1000μg/mL、500μg/mL)或OG高剂量(5×10^(-5)mol/L)联用组、顺铂中剂量(4μg/mL)与仙茅水煎液高、低剂量(1000μg/mL、500μg/mL)或OG高、低剂量(5×10^(-5)mol/L、1×10^(-5)mol/L)联用组较顺铂单用组细胞抑制率均显著升高(P<0.05或P<0.01);顺铂(1.5μg/mL)与仙茅水煎液(1000μg/mL)或OG(5×10^(-5)mol/L)联用组A549/cis细胞的P-gp表达水平均低于顺铂单用组,其中顺铂与仙茅水煎液联用组降低显著(P<0.05)。结论:中药仙茅水煎液及OG均可以降低A549/cis细胞的耐药性,其增敏机制可能与抑制P-gp表达有关。Objective:To explore the effects and underlying mechanism of Curculiginis Rhizoma decoction and orcinol glucoside(OG),the main plasma component,on cisplatin-resistant cells(A549/cis)in non-small cell lung cancer(NSCLC).Methods:The binding affinity of OG to P-glycoprotein(P-gp)was evaluated by AutoDock Vina.CCK-8 assay was used to detect the cell viability inhibition rate of cisplatin,Curculiginis Rhizoma decoction,OG,and their combination on A549/cis cells.The P-gp expression level in A549/cis cells was detected by Western blot.Results:The binding energy of simulated molecular docking of OG to P-gp was-7.6 kcal/mol(1 cal=4.184 J),indicating that they had good binding activity.The half-maximal inhibitory concentration(IC50)of cisplatin on A549/cis cells was about 5.5μg/mL.Curculiginis Rhizoma decoction or OG in the gradient concentrations showed no significant effect on the viability of A549/cis cells.The cell inhibition rates in drug combination groups[low-dose cisplatin(1.5μg/mL)combined with high-and low-dose Curculiginis Rhizoma decoction(1000 and 500μg/mL)or high-dose OG(5×10^(-5)mol/L),and medium-dose cisplatin(4μg/mL)combined with high-and low-dose Curculiginis Rhizoma decoction(1000 and 500μg/mL)or high-and low-dose OG(5×10^(-5)and 1×10^(-5)mol/L)]were higher than that in the cisplatin group(P<0.05 or P<0.01).The P-gp expression levels of A549/cis cells in groups of cisplatin(1.5μg/mL)combined with Curculiginis Rhizoma decoction(1000μg/mL)or OG(5×10^(-5)mol/L)were both lower than that of the cisplatin group,and P-gp expression level in the group of cisplatin combined with Curculiginis Rhizoma decoction lowered significantly(P<0.05).Conclusion:Curculiginis Rhizoma decoction and OG can both reduce the drug resistance of A549/cis cells,and the sensitivity-enhancing mechanism may be related to the inhibition of P-gp expression.

关 键 词：仙茅 苔黑酚葡萄糖苷 非小细胞肺癌 肺癌耐药细胞 P糖蛋白 顺铂 多药耐药 ATP结合盒 

分 类 号：R285.5[医药卫生—中药学]