表木栓醇对人结肠腺癌细胞LS174T中P-糖蛋白基因表达的影响  

Effects of epifriedelanol on gene expression of P-glycoprotein in human colorectal adenocarcinoma cell line LS174T

作　　者：江洁 张小丽 向施佳 姚丽华 钟国平[3] 黄民[3] 黎玉华 JIANG Jie;ZHANG Xiao-li;XIANG Shi-jia;YAO Li-hua;ZHONG Guo-ping;HUANG Min;LI Yu-hua(Department of Pharmacy,Gaoxin Hospital of The First Affiliated Hospital of Nanchang University,Nanchang 330012,Jiangxi Province,China;School of LifeScience,JiangxiScience&Technology Normal University,Nanchang 330013,Jiangxi Province,China;School of Pharmaceutical Sciences,Sun Yat-Sen University,Guangzhou 510006,Guangdong Province,China)

机构地区：[1]南昌大学第一附属医院高新医院药剂科,江西南昌330012 [2]江西科技师范大学生命科学学院,江西南昌330013 [3]中山大学药学院,广东广州510006

出　　处：《中国临床药理学杂志》2025年第1期50-54,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金青年科学基金资助项目(82104285);江西省卫生健康委科技计划基金资助项目(202140141);江西省中医药科技计划基金资助项目(2023B1382)。

摘　　要：目的研究表木栓醇(Epi)对人结肠腺癌细胞LS174T中P-糖蛋白(P-gp)基因表达的影响及机制。方法将LS174T细胞分为对照组和低、中、高剂量实验组。低、中、高剂量实验组分别用5、10、20μmol·L^(-1)Epi处理细胞;对照组用0.1%二甲基亚砜处理细胞。用聚合酶链反应检测Epi对LS174T细胞中P-gPmRNA表达水平的影响,用孕烷X受体(PXR)-多药耐药蛋白1(MDR1/P-gp)双荧光报告基因实验检测Epi对LS174T细胞中MDR1(P-gp)荧光素酶活性的影响,用蛋白质印迹法检测Epi对LS174T细胞中P-gp及核因子-κB(NF-κB)通路相关蛋白表达水平的影响。结果中、高剂量实验组和对照组的P-gPmRNA相对表达水平分别为52.24±5.19、23.00±3.52和100.00±9.00,P-gp蛋白相对表达水平分别为86.37±9.96、74.85±15.92和100.00±12.91,荧光素酶相对活性分别为230.19±41.32、203.10±52.84和279.67±19.20,细胞核内p65(RelA/p65)相对表达水平分别为132.36±23.93、145.96±25.15和100.00±10.88,细胞质中磷酸化NF-κB抑制蛋白激酶α/β(p-IKKα/β)相对表达水平分别为184.00±54.82、290.10±49.59和100.00±15.34,磷酸化NF-κB抑制蛋白α(p-IκBα)相对表达水平分别为125.73±18.77、133.69±20.25和100.00±8.12,IκBα相对表达水平分别为78.36±14.83、70.44±14.57和100.00±22.82。中、高剂量实验组的上述指标与对照组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01,P<0.001)。结论Epi可以抑制人结肠腺癌细胞LS174T中P-gp的基因表达,其机制可能与激活NF-κB通路和抑制PXR途径有关。Objective To investigate the effect of epifriedelanol(Epi)on gene expression of P-glycoprotein(P-gp)in human colorectal adenocarcinoma cell line LS174T and its mechanism.Methods LS174T cells were divided into control group and experimental-L,-M,-H groups.Experimental-L,-M,-H groups were treated with 5,10,20μmol·L^(-1)Epi,respectively.Control group was treated with 0.1% dimethyl sulfoxide.Polymerase chain reaction was used to detect the mRNA expression level of P-gp.The effect of Epi on multidrug resistance protein 1(MDR1/P-gp)luciferase activity was investigated by pregnane X receptor(PXR)-MDR1/P-gp dual luciferase reporter gene assay.In addition,Western Blot was used to detect the protein expression level of P-gp and the nuclear factor-κB(NF-κB)pathway related proteins.Results The relative expression levels of P-gp mRNA in experimental-M,-H groups and control group were 52.24±5.19,23.00±3.52 and 100.00±9.00;the relative expression levels of P-gp protein were 86.37±9.96,74.85±15.92 and 100.00±12.91;the relative activities P-gp luciferase were 230.19±41.32,203.10±52.84 and 279.67±19.20;the relative expression levels of p65(RelA/p65)in nucleus were 132.36±23.93,145.96±25.15 and 100.00±10.88;the relative expression levels of phosphorylation NF-κB inhibits protein kinase α/β(p-IKKα/β)in cytoplasm were 184.00±54.82,290.10±49.59 and 100.00±15.34;the relative expression levels of phosphorylated NF-κB inhibitory protein α(p-IκBα)in cytoplasm were 125.73±18.77,133.69±20.25 and 100.00±8.12;the relative expression levels of IκBα in cytoplasm were 78.36±14.83,70.44±14.57 and 100.00±22.82,respectively.The above indexes of experimental-M and experimental-H groups were compared with control group,and the differences were statistically significant(P<0.05,P<0.01,P<0.001).Conclusion Epi can down-regulate the gene expression of P-gp in human colorectal adenocarcinoma cell line LS174T,and the mechanism may be related to activation of NF-κB and suppression of PXR.

关 键 词：表木栓醇 P-糖蛋白 LS174T细胞 核因子-ΚB 孕烷X受体 

分 类 号：R28[医药卫生—中药学]