新型抗真菌剂TTS-12对白念珠菌麦角甾醇生物合成相关ERG基因表达的作用研究

Study on Regulation of ERG Genes Expression in Candida albicans by a New Anti-Fungi Agent TTS-12

目的研究刺蒺藜(Tribulus terrestris L.)中分离的皂苷类新型抗真菌剂TTS-12对白念珠菌基因表达谱的影响。方法采用微量液基稀释法研究了TTS-12体外MIC80值,并确定合适的作用时间和药物浓度,制备基因芯片,研究了TTS-12对白色念珠菌麦角甾醇生物合成直接相关ERG基因表达的变化,RT-PCR技术验证了芯片结果。结果 TTS-12浓度8μg.mL-1,35℃,250 r.min-1,YNB培养基与SC5314共培养3 h,提取空白菌与药物作用菌mRNA,与白念珠菌芯片杂交;TTS-12作用后,麦角甾醇生物合成直接相关的ACS1、ACS2、ERG1、ERG2、ERG6、ERG7、ERG11、ERG25、ERG26及ERG27基因下调。结论 TTS-12通过与细胞膜上甾醇直接结合,抑制ERG基因表达发挥抗真菌作用。

OBJECTIVE To analyze the regulation effects of TTS-12, a new class of saponins anti-fungal agent which is isolated from Tribulus terrestris L, on the expression of Candida albicans ERG genes. METHODS The incubation time and concentration of TTS-12 were established using broth mierodilution method. DNA microarray analysis was used to investigate the expression changes of TTS-12 target gene ERG related to the ergosterol biosynthesis in Candida albicans. Real-time reverse transcription-PCR was used to confirm microarray results. RESULTS Genes ACS1, ACS2, ERG1, ERG2, ERC,6, ERG7, ERGll, ERG25, ERG26 and ERG27, which directly involved in ergosterol synthesis pathway were depressed. CONCLUSION TTS-12 may function through directly bind- ing to the sterol on the cell membrane and inhibiting the ERG genes expression.