Center for Nanoscience and Nanotechnology, Panjab University, Chandigarh 160014, India.
Department of Microbial Biotechnology, Panjab University (South campus) Sector-25, Chandigarh 160014, India.
J Mycol Med. 2018 Sep;28(3):443-451. doi: 10.1016/j.mycmed.2018.04.008. Epub 2018 May 24.
In the present study, we have evaluated the antifungal activity of the seed, root and leaf of Paeonia emodi (commonly known as Himalayan peony) in four common solvents (acetone, chloroform, methanol and water) against six fungal strains. The methanolic seed extract (MSE) showed promising antifungal activity against Candida albicans (6.25mg/mL), Candida glabrata (3.12mg/mL) and Candida parapsilosis (12.50mg/mL) among all the fungal strains tested. Combination of the MSE with the well-known commercial antifungal drugs amphotericin B (Amp B), nystatin (NYS) and fluconazole (FLC) resulted in the killing of C. glabrata at non-inhibitory concentrations, i.e., 0.35μg/mL for Amp B, 0.55μg/mL for NYS and 1.19μg/mL for FLC. Notably, MSE caused cell wall damage of C. glabrata cells, as confirmed by confocal microscopy, flowcytometry and scanning electron microscopy (SEM). The MSE was fractionated by thin layer chromatography (TLC). TLC-bioautography was used to determine the active compounds present in the MSE. Column chromatography was used to separate the potential active compounds from the MSE. Furthermore, gas chromatography-mass spectrometry (GC-MS) andfourier-transform infrared spectroscopy (FTIR) were used to identify the phytocomponents of the MSE. These experiments revealed 13-docosenamide/9-octadecenamide/trans-13-docosenamide (89.70%) as being the predominant compound using a chloroform/methanol solvent system for the separation. Interestingly, the MSE also exhibited less significant cytotoxicity at the minimum inhibitory concentration (MIC) against mammalian cells (HeLa and HEK293). This study suggests that the MSE of P. emodi can be used for the treatment of C. glabrata infection.
在本研究中，我们评估了青藏高原獐牙菜（俗称喜马拉雅牡丹）的种子、根和叶在四种常见溶剂（丙酮、氯仿、甲醇和水）中的抗真菌活性，针对六种真菌菌株。在所有测试的真菌菌株中，甲醇种子提取物（MSE）对白色念珠菌（Candida albicans）（6.25mg/mL）、光滑念珠菌（Candida glabrata）（3.12mg/mL）和近平滑念珠菌（Candida parapsilosis）（12.50mg/mL）表现出有希望的抗真菌活性。MSE 与知名的商业抗真菌药物两性霉素 B（Amp B）、制霉菌素（NYS）和氟康唑（FLC）联合使用，可在非抑制浓度下杀死 C. glabrata，即 Amp B 为 0.35μg/mL，NYS 为 0.55μg/mL，FLC 为 1.19μg/mL。值得注意的是，MSE 导致 C. glabrata 细胞的细胞壁损伤，这一点通过共聚焦显微镜、流式细胞术和扫描电子显微镜（SEM）得到证实。MSE 通过薄层层析（TLC）进行分离。TLC 生物自显影用于确定 MSE 中存在的活性化合物。柱层析用于从 MSE 中分离潜在的活性化合物。此外，气相色谱-质谱联用（GC-MS）和傅里叶变换红外光谱（FTIR）用于鉴定 MSE 的植物成分。这些实验使用氯仿/甲醇溶剂系统分离，发现 13-二十二酰胺/9-十八烯酰胺/反式-13-二十二酰胺（89.70%）是主要化合物。有趣的是，MSE 在抗真菌最低抑制浓度（MIC）下对哺乳动物细胞（HeLa 和 HEK293）的细胞毒性也较小。本研究表明，青藏高原獐牙菜的 MSE 可用于治疗 C. glabrata 感染。
