The First Affiliated Hospital and Institute of Mycology of Jinan University, Guangzhou, Guangdong, China.
Pharm Biol. 2013 Jun;51(6):749-52. doi: 10.3109/13880209.2013.764537. Epub 2013 Mar 25.
Our previous study demonstrated that tetrandrine (TET) could reverse the resistance of Candida albicans to fluconazole.
The aim of this study was to investigate the molecular mechanism underlying this action.
Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) was performed to compare the expression levels of the drug resistance genes CDR1, CDR2, MDR1, FLU1 and ERG11 in fluconazole-sensitive CA-3 and resistant CA-16 cells that were either treated with FLC and/or TET or left as untreated controls. In addition, intracellular ATP levels were measured using an ATP assay kit, and the expression level of the energy metabolism gene ADH1 was measured by real-time RT-PCR.
Compared with FLC/TET-free conditions, FLC + TET treatment strains showed statistically different (p < 0.05) expression of CDR1 and CDR2 (increased in the FLC-sensitive strains, while decreased in the FLC-resistant strains), MDR1 (increased in the FLC-resistant strains), FLU1 and ERG11 (increased in the FLC-sensitive strains), ADH1 (decreased in both the FLC-sensitive and the FLC-resistant strains). And also, the FLC + TET treatment decreased the intracellular ATP levels in both the FLC-sensitive and the FLC-resistant strains (p < 0.05).
These results suggest that changes in the expression levels of the drug resistance genes CDR1 and CDR2, the cellular ATP supply and the expression level of the energy metabolism gene ADH1 contribute to the TET-mediated reversal of the fluconazole resistance of C. albicans.
我们之前的研究表明汉防己甲素（TET）可以逆转白色念珠菌对氟康唑的耐药性。
本研究旨在探讨其作用的分子机制。
采用实时逆转录聚合酶链反应（real-time RT-PCR）比较氟康唑敏感 CA-3 株和耐药 CA-16 株在分别用 FLC 和/或 TET 处理或未处理的情况下，耐药基因 CDR1、CDR2、MDR1、FLU1 和 ERG11 的表达水平。此外，用 ATP 测定试剂盒测量细胞内 ATP 水平，并用 real-time RT-PCR 测量能量代谢基因 ADH1 的表达水平。
与 FLC/TET 无处理条件相比，FLC+TET 处理菌株的 CDR1 和 CDR2（在 FLC 敏感菌株中增加，而在 FLC 耐药菌株中减少）、MDR1（在 FLC 耐药菌株中增加）、FLU1 和 ERG11（在 FLC 敏感菌株中增加）、ADH1（在 FLC 敏感和 FLC 耐药菌株中均减少）的表达存在统计学差异（p<0.05）。并且，FLC+TET 处理降低了 FLC 敏感和 FLC 耐药菌株的细胞内 ATP 水平（p<0.05）。
这些结果表明，耐药基因 CDR1 和 CDR2 的表达水平变化、细胞内 ATP 供应以及能量代谢基因 ADH1 的表达水平变化可能导致 TET 介导的白色念珠菌氟康唑耐药性逆转。
