酒川芎联合吉非替尼或厄洛替尼治疗EGFR突变型非小细胞肺癌脑移植瘤的作用机制  

Mechanism of Wine-processed Chuanxiong Rhizoma Combined with Gefitinib or Erlotinib in Treating EGFR Mutant NSCLC Metastatic Encephaloma

作　　者：荣翔宇 赵倩 张振 周星琦 李姗姗 李娴 RONG Xiangyu;ZHAO Qian;ZHANG Zhen;ZHOU Xingqi;LI Shanshan;LI Xian(School of Pharmacy,Bengbu Medical University/Anhui Biochemical and Pharmaceutical Engineering Technology Research Center,Bengbu Anhui China 233030)

机构地区：[1]蚌埠医科大学药学院/安徽省生化药物工程技术研究中心,安徽蚌埠233030

出　　处：《中医学报》2024年第7期1534-1542,共9页Acta Chinese Medicine

基　　金：安徽省重点研究与开发计划项目(202104g01020017);安徽省生化工程中心科研平台开放课题项目(2023SYKFD05)。

摘　　要：目的:探讨酒川芎(wine-processed Chuanxiong Rhizoma,WCR)联合吉非替尼或厄洛替尼治疗表皮生长因子受体(epidermal growth factor receptor,EGFR)突变型非小细胞肺癌(non-small cell lung cancer,NSCLC)脑移植瘤的作用机制。方法:建立ABCB1-MDCK细胞和PC9细胞共培养体系,采用流式细胞术检测WCR(2 g·L^(-1))联合吉非替尼或厄洛替尼(10μmol·L^(-1)、20μmol·L^(-1))作用于PC9细胞的凋亡情况。通过ABCB1-MDCK细胞单层转运模型评估WCR(0.5 g·L^(-1)、1 g·L^(-1)、2 g·L^(-1))联合吉非替尼或厄洛替尼(8μmol·L^(-1))的跨膜转运情况,并计算表观渗透系数(Papp)。体外培养ABCB1-MDCK细胞,采用透射电镜法、免疫荧光法(immunofluorescence,IF)和蛋白质免疫印迹法(Western blot,WB)检测超微结构和紧密连接蛋白-1(zonula occludens^(-1),ZO-1)、P-糖蛋白(P-glycoprotein,P-gp)表达水平。建立裸鼠NSCLC脑移植瘤模型,检测WCR联合吉非替尼或厄洛替尼对裸鼠脑内肿瘤生长、生存时间和药物含量的影响。结果:与单用吉非替尼或厄洛替尼比较,WCR联合吉非替尼或厄洛替尼可增加PC9细胞的凋亡率(P<0.05)。WCR联合吉非替尼或厄洛替尼的Papp(AP→BL)较吉非替尼或厄洛替尼增加(P<0.05),且与WCR浓度呈正相关。透射电镜结果显示,WCR可破坏ABCB1-MDCK细胞的紧密连接结构。IF和WB结果显示,WCR可降低ABCB1-MDCK细胞的ZO-1、P-gp表达水平(P<0.01)。WCR联合吉非替尼或厄洛替尼可抑制脑内肿瘤生长、延长裸鼠生存时间并增加吉非替尼或厄洛替尼含量。结论:WCR联合吉非替尼或厄洛替尼可能通过促进细胞凋亡、改善药物转运以及下调ZO-1、P-gp表达水平,从而发挥治疗EGFR突变型NSCLC脑移植瘤的作用。Objective:Explore the mechanism of wine-processed Chuanxiong Rhizoma(WCR)combined with Gefitinib or Erlotinib in treating epidermal growth factor receptor(EGFR)mutant ions in non-small cell lung cancer non-small cell lung cancer(NSCLC)metastatic encephaloma.Methods:The co-culture system of ABCB1-MDCK cells and PC9 cells was established,and the apoptosis of PC9 cells treated with WCR(2 g·L^(-1))combined with Gefitinib or Erlotinib was detected with flow cytometry.The transmembrane transport of WCR(0.5 g·L^(-1),1 g·L^(-1),2 g·L^(-1))combined with Gefitinib or Erlotinib(8μmol·L^(-1))was evaluated with ABCB1-MDCK cell monolayer transport model,and the apparent permeability coefficient(Papp)was calculated.ABCB1-MDCK cells were cultured in vitro.Transmission electron microscopy,immunofluorescence(IF)and western blot(WB)were used to detect the ultrastructure of ABCB1-MDCK cells and zonula occludens^(-1)(ZO-1)protein and P-glycoprotein(P-gp)expression levels.A nude mouse model of NSCLC metastatic encephaloma was established.The effects of WCR combined with Gefitinib or Erlotinib on tumor growth,survival time and drug content in nude mice were detected.Results:Compared with Gefitinib or Erlotinib alone,WCR combined with Gefitinib or Erlotinib significantly increased the apoptosis rate of PC9 cells significantly(P<0.05),and Papp(A→B)of WCR combined with Gefitinib or Erlotinib was significantly higher(P<0.05),which was positively correlated with the concentration of WCR.It was showed in transmission electron microscopy that WCR could destroy the tight junction structure of ABCB1-MDCK cells.IF and WB results suggested that WCR could reduce the expression levels of ZO-1 and P-gp in ABCB1-MDCK cells significantly(P<0.01).WCR combined with Gefitinib or Erlotinib can inhibit the growth of brain tumors,prolong the survival time of nude mice and increase the content of Gefitinib or Erlotinib.Conclusion:WCR combined with Gefitinib or Erlotinib may play a role in the treatment of EGFR-mutant NSCLC metastatic encephaloma by p

关 键 词：非小细胞肺癌 脑移植瘤 酒川芎 吉非替尼 厄洛替尼 EGFR 血脑屏障 ZO-1 小鼠 

分 类 号：R285.5[医药卫生—中药学]