Kitiyanant Vorawat, Lotrakul Pongtharin, Kanchanabanca Chompoonik, Padungros Panuwat, Punnapayak Hunsa, Prasongsuk Sehanat, Chanvorachote Pithi
Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
Plant Biomass Utilization Research Unit, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.
In Vivo. 2019 Mar-Apr;33(2):425-432. doi: 10.21873/invivo.11490.
BACKGROUND/AIM: Oxidative stress caused by the production of excessive cellular reactive oxygen species (ROS) and high levels of nitric oxide contribute to several human pathologies. This study aimed to examine the anti-oxidant effects of fusigen, a compound produced from Aureobasidium melanogenum.
Extracts of A. melanogenum were selected as a source for the isolation of fusigen. The anti-oxidant, nitric oxide suppression, as well as the free radical scavenging activities of fusigen were tested in BEAS-2B human bronchial epithelial cell line (BEAS-2B cells) and human dermal papilla cells (DP cells) using specific fluorescence dyes and flow cytometry analysis. Cell viability was determined by the MTT assay.
Fusigen did not exert cytotoxicity in the human normal BEAS-2B and DP cells at concentrations up to 100 μM. Fusigen decreased basal levels of cellular ROS, as well as the levels of ROS induced by hydrogen peroxide and ferrous ion enrichment. ROS decreasing effect was confirmed in DP cells. In addition, fusigen treatment suppressed intracellular NO levels in both BEAS-2B and DP cells.
The optimal process of production of purified fusigen from A. melanogenum was determined. Fusigen exhibited a low cytotoxic effect and the potential to suppress ROS and NO. These results demonstrated that fusigen may be used for the treatment or prevention of human diseases.
背景/目的：细胞内活性氧（ROS）生成过多以及一氧化氮水平升高所导致的氧化应激会引发多种人类疾病。本研究旨在考察由黑曲霉产生的一种化合物fusigen的抗氧化作用。
选用黑曲霉提取物作为分离fusigen的来源。利用特定荧光染料和流式细胞术分析，在BEAS-2B人支气管上皮细胞系（BEAS-2B细胞）和人真皮乳头细胞（DP细胞）中检测fusigen的抗氧化、一氧化氮抑制以及自由基清除活性。通过MTT法测定细胞活力。
在浓度高达100μM时，fusigen对人正常BEAS-2B细胞和DP细胞未表现出细胞毒性。Fusigen降低了细胞内ROS的基础水平，以及由过氧化氢和亚铁离子富集诱导产生的ROS水平。在DP细胞中证实了ROS降低作用。此外，fusigen处理抑制了BEAS-2B细胞和DP细胞内的NO水平。
确定了从黑曲霉中生产纯化fusigen的最佳工艺。Fusigen表现出低细胞毒性作用以及抑制ROS和NO的潜力。这些结果表明，fusigen可用于治疗或预防人类疾病。
