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首页> 外文期刊>Biochemistry and molecular biology education >Laboratory Exercises; Using Paraquat to Generate Anion Free Radicals and Hydrogen Peroxide in In Vitro: Antioxidant Effect of Vitamin E; A PROCEDURE TO TEACH THEORETICAL AND EXPERIMENTAL PRINCIPLES OF REACTIVE OXYGEN SPECIES BIOCHEMISTRY
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Laboratory Exercises; Using Paraquat to Generate Anion Free Radicals and Hydrogen Peroxide in In Vitro: Antioxidant Effect of Vitamin E; A PROCEDURE TO TEACH THEORETICAL AND EXPERIMENTAL PRINCIPLES OF REACTIVE OXYGEN SPECIES BIOCHEMISTRY

机译:实验室练习;使用百草枯体外产生阴离子自由基和过氧化氢:维生素E的抗氧化作用;活性氧生物化学理论和实验原理的教学方法

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摘要

The theoretical basis of reactive oxygen species and their impact on health issues are relatively easy to understand by biomedical students. The detection of reactive oxygen species requires expensive equipment, the procedures are time consuming and costly, and the results are hard to interpret. Moreover, cause-and-effect relationships in the living system are not so evident. In this report, we adapted a two-step procedure to detect anion superoxide radicals and hydrogen peroxide generation in lymphocytes exposed to paraquat by using nitroblue tetrazolium salt and dihydrorhodamine, respectively. Also, a two-step assay was performed to evaluate lymphocyte viability and nuclei morphologic changes on paraquat exposure for 1 and 24 hours incubation time by using trypan blue exclusion assay and acridine orange/ ethidium bromide staining technique, respectively. Vitamin E was used as antioxidant to inhibit the deleterious effects of paraquat on cells. Students learned how to (i) design and perform experiments in the laboratory, (ii) read critical scientific literature, and (iii) discuss and contrast relevant information about reactive oxygen species as causative agents of cell death phenomenon.
机译:生物医学专业的学生相对容易理解活性氧的理论基础及其对健康问题的影响。活性氧种类的检测需要昂贵的设备,该过程既耗时又昂贵,并且结果难以解释。而且,生命系统中的因果关系并不是那么明显。在本报告中,我们采用了两步法,分别通过使用硝基蓝四唑盐和二氢罗丹明检测暴露于百草枯的淋巴细胞中的阴离子超氧自由基和过氧化氢的产生。此外,通过使用锥虫蓝排除法和a啶橙/溴化乙锭染色技术,分别进行了两步分析以评估百草枯暴露1和24小时后淋巴细胞的活力和细胞核形态变化。维生素E被用作抗氧化剂来抑制百草枯对细胞的有害作用。学生学习了如何(i)在实验室中设计和进行实验,(ii)阅读重要的科学文献,以及(iii)讨论并对比了有关活性氧作为细胞死亡现象病因的相关信息。

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