In Vitro and In Vivo Activities of Pterostilbene against Candida albicans Biofilms
In Vitro and In Vivo Activities of Pterostilbene against Candida albicans Biofilms
复制标题
紫檀芪对抗白色念珠菌生物膜的体外和体内活性
DOI：
10.1128/aac.01583-13
复制
发表时间：
2014-04-01
影响因子：
4.9
通讯作者：
Jiang, Yuan-Ying
中科院分区：
文献类型：
--
作者：
Li, De-Dong;Zhao, Lan-Xue;Jiang, Yuan-Ying
摘要紫檀芪（PTE）是一种来源于多种天然植物的芪类植物抗毒素。在这项研究中，我们评估了PTE对白色念珠菌生物膜的活性，并探讨了潜在的机制。在2，3-双-（2-甲氧基-4-硝基-5-磺基苯基）-2H-四唑-5-甲酰苯胺（XTT）还原试验、生物膜生物量测量、激光共聚焦扫描显微镜和扫描电子显微镜中，我们发现≤16 μg/ml的PTE对C.对白色念珠菌生物被膜有明显的抑制作用，而对嗜酸性念珠菌无抑制作用。提示PTE具有独特的细胞膜效应。结果发现，PTE可抑制生物膜的形成，破坏成熟生物膜的维持。在4 μg/ml时，PTE降低细胞表面疏水性（CSH），抑制菌丝形成。基因表达谱芯片和实时荧光定量RT-PCR显示，暴露于C。16 μg/ml PTE改变了在形态转变、麦角固醇生物合成、氧化还原酶活性、细胞表面和蛋白质解折叠过程（热休克蛋白）中起作用的基因的表达。PTE处理后，微丝相关基因，尤其是Ras/cAMP通路调控的基因，包括ECE 1、ALS 3、HWP 1、HGC 1和RAS 1本身表达下调，表明PTE的微丝膜效应与Ras/cAMP通路有关。然后，我们发现，加入外源性cAMP逆转PTE诱导的丝状生长缺陷。最后，通过大鼠中心静脉导管感染模型，证实了PTE在体内抗C.白色念珠菌生物膜总体而言，PTE对C.白念珠菌生物膜在体外和体内，这些活动与Ras/cAMP途径。
ABSTRACT Pterostilbene (PTE) is a stilbene-derived phytoalexin that originates from several natural plant sources. In this study, we evaluated the activity of PTE against Candida albicans biofilms and explored the underlying mechanisms. In 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction assays, biofilm biomass measurement, confocal laser scanning microscopy, and scanning electron microscopy, we found that ≤16 μg/ml PTE had a significant effect against C. albicans biofilms in vitro, while it had no fungicidal effect on planktonic C. albicans cells, which suggested a unique antibiofilm effect of PTE. Then we found that PTE could inhibit biofilm formation and destroy the maintenance of mature biofilms. At 4 μg/ml, PTE decreased cellular surface hydrophobicity (CSH) and suppressed hyphal formation. Gene expression microarrays and real-time reverse transcription-PCR showed that exposure of C. albicans to 16 μg/ml PTE altered the expression of genes that function in morphological transition, ergosterol biosynthesis, oxidoreductase activity, and cell surface and protein unfolding processes (heat shock proteins). Filamentation-related genes, especially those regulated by the Ras/cyclic AMP (cAMP) pathway, including ECE1, ALS3, HWP1, HGC1, and RAS1 itself, were downregulated upon PTE treatment, indicating that the antibiofilm effect of PTE was related to the Ras/cAMP pathway. Then, we found that the addition of exogenous cAMP reverted the PTE-induced filamentous growth defect. Finally, with a rat central venous catheter infection model, we confirmed the in vivo activity of PTE against C. albicans biofilms. Collectively, PTE had strong activities against C. albicans biofilms both in vitro and in vivo, and these activities were associated with the Ras/cAMP pathway.