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内生真菌 Chaetomium globosum JN711454 的生物评价及其作为药物发现潜在候选物的研究

Biological evaluation of endophytic fungus, Chaetomium globosum JN711454, as potential candidate for improving drug discovery.

机构信息

Department of Chemistry of Natural and Microbial Products, National Research Center, Dokki, Cairo, 12622, Egypt.

出版信息

Cell Biochem Biophys. 2014 Jan;68(1):67-82. doi: 10.1007/s12013-013-9695-4.

DOI:10.1007/s12013-013-9695-4
PMID:23775636
Abstract

The main objective of this research work focused on investigating the biological and chemical aspects of endophytic fungus Chaetomium globosum, for pharmaceutical purposes to improve the drug discovery process. The endophytic C. globosum was isolated from healthy leaves of Egyptian medicinal plant Adiantum capillus-veneris collected from Saint Katherine Protectorate, Sinai, Egypt. The identification of C. globosum was on the basis of classical and molecular taxonomy. Gene encoding for 18S rRNA was partially sequenced, submitted to the GenBank and got the accession number JN711454, to resolve the phylogenetic relations with fungal ancestor using phylogenetic tree. To explore the biosynthetic power of endophytic C. globosum JN711454, the fungus was cultivated over five different media, oatmeal, rice, yeast malt glucose, potato dextrose agar (PDA) and Czapek's dox media, for 3 weeks at 30 °C, followed by extraction with different solvents, ethyl acetate (EA), and methanol. The ethyl acetate extract of C. globosum cultivated on PDA medium was the most potent extract. It showed strong antioxidant activity with EC50 11.5 μg/ml, potent anticancer activity with 55 % toxicity toward HepG-2 cells at 100 μg/ml and 66 % cytotoxicity to FGC4 cells at 250 μg/ml, promising butyrylcholinesterase inhibitory activities (>85 %), and moderate antimicrobial and stopped the attachment of HSV-2 virus to VERO cells. The metabolomic profiling of PDA-EA extract using LC-MS revealed the presence of several metabolites to which the observed bioactivities could be attributed. Here we report for the first time inhibitory activity of endophytic C. globosum JN711454 secondary metabolites to butyrylcholinesterase, one of neuro hydrolase enzymes that play a major role in development of Alzheimer's disease.

摘要

本研究工作的主要目的是研究内生真菌 Chaetomium globosum 的生物学和化学方面,以用于制药目的,从而改进药物发现过程。内生 C. globosum 是从埃及药用植物 Adiantum capillus-veneris 的健康叶片中分离出来的,这些叶片是从埃及圣凯瑟琳保护区的西奈收集的。C. globosum 的鉴定是基于经典和分子分类学。部分测序了编码 18S rRNA 的基因,并提交给 GenBank,获得了注册号 JN711454,以使用系统发育树来解决与真菌祖先的系统发育关系。为了探索内生 C. globosum JN711454 的生物合成能力,将真菌在五种不同的培养基上培养,燕麦、大米、酵母麦芽葡萄糖、土豆葡萄糖琼脂(PDA)和察氏培养基,在 30°C 下培养 3 周,然后用不同的溶剂提取,乙酸乙酯(EA)和甲醇。在 PDA 培养基上培养的 C. globosum 的乙酸乙酯提取物是最有效的提取物。它具有很强的抗氧化活性,EC50 为 11.5μg/ml,对 HepG-2 细胞的毒性为 55%,在 100μg/ml 时对 FGC4 细胞的细胞毒性为 66%,对丁酰胆碱酯酶具有很强的抑制活性(>85%),对微生物有中等抑制作用,并阻止了 HSV-2 病毒附着在 VERO 细胞上。使用 LC-MS 对 PDA-EA 提取物进行代谢组学分析显示,存在几种可能与观察到的生物活性相关的代谢物。在这里,我们首次报道了内生 C. globosum JN711454 次生代谢物对丁酰胆碱酯酶的抑制活性,丁酰胆碱酯酶是神经水解酶之一,在阿尔茨海默病的发展中起主要作用。

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