The chemical attractant against T.virax from pollen grains

花粉粒中针对 T.virax 的化学引诱剂

• 批准号: 04044139
• 负责人: OGIHARA Yukio
• 金额: $ 3.65万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04044139/
• 关键词: Trypanosoma; T.vivax(IL1392); T.bruci gambiense(Wellcome Strain; Anti-Trypanosoma activity; Gallic acid; Cytotoxic activity; T.vivax(IL1392); Fuchsia hybrida; 花粉; 走化反応

Trypanosoma is well known to cause african sleeping sickness in cow and human. The objective of our research focused on the development of anti-trypanosoma drug. We therefore tried to clarify the mechanism of the transformation from fission type to non-fission type by extracting, isolating and determining the structure of chemoattractants for trypasnosoma from pollen. Furthermore, we searched for compounds with anti-trypanosoma activity (cytotoxic activity or growth inhibitory activity) in medicinal plants.1. We first assayed chemoattractive activity of pollen collected from eight plants using trypanosoma vivax. Among them, the crushed pollens from fukusia, hiougi, insiahamayu, chousenasagao showed chemoattractive activity. The water extract of fukusia showed especially strong activity and the methanol extract also showed the activity though weak activity. The water extract further represented erythrocyte agglutination activity. Those activities of water extract of fukusia pollen were found to be in the fraction with molecular weight from 5000 to 1000. We are now undertaking to isolate compounds with those activities.2. We found that gallic acid, 3,4,5-hydroxy-benzoic acid, showed strong anti-trypanosoma activity in vitro assay system using trypanosoma brucei bricei IL Tat 1.4 clone. Its activity was cytotoxic activity but not growth inhibitory activity. Gallic acid at a concentration of 10mug/ml killed the protozoan within 2 hours, at 6mug/ml within 6 hours. This strong cytotoxic activity depended on the concentration of protozoan in culture. That is to say, trypanosoma less than concentration of 10^6/ml was killed by 10 mug/ml of gallic acid. Morphological study indicated that the death of trypanosoma was a kind of apoptosis, although DNA fragmentation characteristic of apoptotic death was not detected. We are further investigating the mechanism by which gallic acid shows cytotoxic activity.

锥虫是众所周知的导致非洲昏睡病牛和人类。本研究的目的是开发抗锥虫药物。因此，我们试图从花粉中提取、分离和测定锥虫化学引诱物的结构，以阐明锥虫由分裂型向非分裂型转化的机制。此外，我们还在药用植物中寻找具有抗锥虫活性（细胞毒活性或生长抑制活性）的化合物。我们首先用间日锥虫测定了从八种植物中收集的花粉的化学吸引活性。其中，fukusia、hiougi、insiahamayu、chousenasagao等植物的花粉具有化学引诱活性。其中，福古草的水提物活性最强，甲醇提取物虽然活性较弱，但也有活性。水提物进一步表现出红细胞凝集活性。这些活性物质主要存在于分子量为5000 ~ 1000的组分中。我们现在正着手分离具有这些活性的化合物。利用布氏锥虫IL达特1.4克隆体外抗锥虫活性测定系统，发现没食子酸即3，4，5-羟基苯甲酸具有较强的抗锥虫活性。其活性为细胞毒活性，但不具有生长抑制活性。没食子酸浓度为10 μ g/ml时在2小时内杀死原虫，浓度为6 μ g/ml时在6小时内杀死原虫。这种强烈的细胞毒活性依赖于培养物中原生动物的浓度。也就是说，浓度低于10^6/ml的锥虫被10 μ g/ml的没食子酸杀死。形态学研究表明，锥虫的死亡是一种细胞凋亡，但未检测到凋亡性DNA断裂。我们正在进一步研究没食子酸显示细胞毒活性的机制。