Mechanism of action of tetrandrine, a natural inhibitor of Candida albicans drug efflux pumps.
Synergistic effects have previously been observed for a natural compound, tetrandrine (TET), with fluconazole (FLC) in vitro and in the treatment of Candida albicans-infected mice. To investigate the mechanisms of these synergistic effects, 16 strains of C. albicans from the same parent but with different FLC sensitivities were examined using flow cytometry and fluorescent spectrophotometry. Rhodamine 123 (Rh123)-positive cells and intracellular Rh123 fluorescence intensity were determined in accumulation/efflux experiments involving no or a noncytotoxic dose of TET. Total RNA extracted from each strain was used to compare the expressions of drug efflux pump genes in FLC-susceptible, -susceptible dose-dependent, and -resistant strains before and 24 h after TET administration. Accumulation experiments determined that mean percentages of Rh123-positive cells were 26.65% (TET-free) and 70.99% (TET 30 microg/ml), and mean respective intracellular Rh123 fluorescence intensities were 11.34 and 18.00. Efflux experiments showed that percentages of Rh123-positive cells were 1.79% (TET free) and 42.57% (TET 30 microg/ml), respectively, and respective mean intracellular Rh123 fluorescence intensities were 0.74 and 2.19. Differences in MDR1, FLU1, CDR1, and CDR2 expression levels in the absence of TET were statistically significant (p<0.05) between FLC-susceptible, -susceptible dose-dependent, and -resistant strains. Compared with TET-free conditions, 24 h TET-treated strains showed statistically different (p<0.05) expression of MDR1 (FLC-resistant strain), FLU1 (FLC-susceptible dose-dependent and -resistant strains), and CDR1 and CDR2 (FLC-susceptible, -susceptible dose-dependent, and -resistant strains). Thus TET can inhibit the C. albicans drug efflux system and reduce drug efflux. Its mechanism of action is related to the inhibition of expression of the drug efflux pump genes MDR1, FLU1, CDR1, and CDR2.
Zhang
Hong
H
Department of Dermatology, The First Affiliated Hospital, Jinan University, Guangzhou, China. tzhangh@jnu.edu.cn
Gao
Aili
A
Li
Fengxia
F
Zhang
Gehua
G
Ho
Hon In
HI
Liao
Wanqing
W
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000235:genetics; D018528:ATP-Binding Cassette Transporters / Q000037:antagonists & inhibitors* / Q000235:genetics; D000818:Animals; D000935:Antifungal Agents / Q000378:metabolism* / Q000494:pharmacology*; D044182:Benzylisoquinolines / Q000494:pharmacology*; D002176:Candida albicans / Q000187:drug effects* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D018432:Drug Resistance, Multiple / Q000235:genetics; D004357:Drug Synergism; D015725:Fluconazole / Q000378:metabolism* / Q000494:pharmacology*; D005656:Fungal Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D026901:Membrane Transport Proteins / Q000235:genetics; D051379:Mice; D012331:RNA, Fungal / Q000378:metabolism
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents; D044182:Benzylisoquinolines; C094693:CDR1 protein, Candida albicans; C417782:FLU1 protein, Candida albicans; D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D012331:RNA, Fungal; C009438:tetrandrine; D015725:Fluconazole