title: Improvement of Transmembrane Transport Mechanism Study of Imperatorin on P-Glycoprotein-Mediated Drug Transport.
abstract: P-glycoprotein (P-gp) affects the transport of many drugs; including puerarin and vincristine. Our previous study demonstrated that imperatorin increased the intestinal absorption of puerarin and vincristine by inhibiting P-gp-mediated drug efflux. However; the underlying mechanism was not known. The present study investigated the mechanism by which imperatorin promotes P-gp-mediated drug transport. We used molecular docking to predict the binding force between imperatorin and P-gp and the effect of imperatorin on P-gp activity. P-gp efflux activity and P-gp ATPase activity were measured using a rhodamine 123 (Rh-123) accumulation assay and a Pgp-Glo™ assay; respectively. The fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH) was used to assess cellular membrane fluidity in MDCK-MDR1 cells. Western blotting was used to analyze the effect of imperatorin on P-gp expression; and P-gp mRNA levels were assessed by qRT-PCR. Molecular docking results demonstrated that the binding force between imperatorin and P-gp was much weaker than the force between P-gp and verapamil (a P-gp substrate). Imperatorin activated P-gp ATPase activity; which had a role in the inhibition of P-gp activity. Imperatorin promoted Rh-123 accumulation in MDCK-MDR1 cells and decreased cellular membrane fluidity. Western blotting demonstrated that imperatorin inhibited P-gp expression; and qRT-PCR revealed that imperatorin down-regulated P-gp (MDR1) gene expression. Imperatorin decreased P-gp-mediated drug efflux by inhibiting P-gp activity and the expression of P-gp mRNA and protein. Our results suggest that imperatorin could down-regulate P-gp expression to overcome multidrug resistance in tumors.
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. lyzlyg@163.com.
authors.forename: Zheng-Gen
authors.initials: ZG
authors.lastname: Liao
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. tangtao12306@sina.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Tang
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. guanxj52611905@sina.com.
authors.forename: Xue-Jing
authors.initials: XJ
authors.lastname: Guan
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. sober96@foxmail.com.
authors.forename: Wei
authors.initials: W
authors.lastname: Dong
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. evens_zhang@163.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. weiweihaoyunqi@163.com.
authors.forename: Guo-Wei
authors.initials: GW
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. doscat@163.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.affiliation: Key Laboratory of Modern Preparation of TCM, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, China. marryrose1013@163.com.
authors.forename: Xin-Li
authors.initials: XL
authors.lastname: Liang
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D029969:Angelica / Q000737:chemistry; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D004161:Diphenylhexatriene / Q000737:chemistry; D004285:Dogs; D011564:Furocoumarins / Q000494:pharmacology*; D007529:Isoflavones / Q000378:metabolism*; D061985:Madin Darby Canine Kidney Cells; D062105:Molecular Docking Simulation; D028321:Plant Preparations / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis; D014750:Vincristine / Q000378:metabolism*
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D011564:Furocoumarins; D007529:Isoflavones; D028321:Plant Preparations; D012333:RNA, Messenger; D004161:Diphenylhexatriene; D014750:Vincristine; C031534:imperatorin; C033607:puerarin
keywords: P-glycoprotein (P-gp); imperatorin; mechanism; transmembrane transport