Name: production and detection of reactive oxygen species (ros) in cancers
Uploaded: 2011-11-21T13:00:00
Description: Watch this Scientific Journal Video about Production and Detection of Reactive Oxygen Species ROS in Cancers at JoVE.com

这项工作是支持由国立卫生研究院（CA142664）。

1。 ROS的检测，使用羧基- H 2 DCFDA 羧基- H 2 DCFDA非荧光，但是，当这种试剂氧化的活性氧的存在，成为绿色荧光。 <ol><li>立即使用前，准备在无菌二甲基亚砜（DMSO）或100％的乙醇的新鲜原液的羧基- H 2 DCFDA 。避免你的染料多个解冻/冻结周期。 </li><li>洗净细胞，与HEPES缓冲盐溶液（HBSS）或磷酸盐缓冲液（PBS）删除原介质的痕迹。 </li><li>负载与染料的?...

<ol>
	<li>Guzik, T. J., Korbut, R., Adamek-Guzik, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nitric+oxide+and+superoxide+in+inflammation+and+immune+regulation.">Nitric oxide and superoxide in inflammation and immune regulation.</a> J Physiol Pharmacol. 54, 469-487 (2003).</li><li>Perl, A., Gergely, P., Banki, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+dysfunction+in+T+cells+of+patients+with+systemic+lupus+erythematosus.">Mitochondrial dysfunction in T cells of patients with systemic lupus erythematosus.</a> Int Rev Immunol. 23, 293-313 (2004).</li><li>Valko, M., Leibfritz, D., Moncol, J., Cronin, M. T., Mazur, M., Telser, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free+radicals+and+antioxidants+in+normal+physiological+functions+and+human+disease.">Free radicals and antioxidants in normal physiological functions and human disease.</a> Int J Biochem Cell Biol. 39, 44-84 (2007).</li><li>Droge, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free+radicals+in+the+physiological+control+of+cell+function.">Free radicals in the physiological control of cell function.</a> Physiol Rev. 82, 47-95 (2002).</li><li>Nakamura, K., Yube, K., Miyatake, A., Cambier, J. C., Hirashima, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+CD4+D3-D4+membrane+proximal+extracellular+domain+for+the+inhibitory+effect+of+oxidative+stress+on+activation-induced+CD4+down-regulation+and+its+possible+role+for+T+cell+activation.">Involvement of CD4 D3-D4 membrane proximal extracellular domain for the inhibitory effect of oxidative stress on activation-induced CD4 down-regulation and its possible role for T cell activation.</a> Mol Immunol. 39, 909-921 (2003).</li><li>Los, M., Droge, W., Stricker, K., Baeuerle, P. A., Schulze-Osthoff, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrogen+peroxide+as+a+potent+activator+of+T+lymphocyte+functions.">Hydrogen peroxide as a potent activator of T lymphocyte functions.</a> Eur J Immunol. 25, 159-165 (1995).</li><li>Deem, T. L., Cook-Mills, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+cell+adhesion+molecule+1+(VCAM-1)+activation+of+endothelial+cell+matrix+metalloproteinases:+role+of+reactive+oxygen+species.">Vascular cell adhesion molecule 1 (VCAM-1) activation of endothelial cell matrix metalloproteinases: role of reactive oxygen species.</a> Blood. 104, 2385-2393 (2004).</li><li>Pacher, P., Beckman, J. S., Liaudet, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nitric+oxide+and+peroxynitrite+in+health+and+disease.">Nitric oxide and peroxynitrite in health and disease.</a> Physiol Rev. 87, 315-424 (2007).</li><li>Griendling, K. K., Sorescu, D., Lassegue, B., Ushio-Fukai, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+protein+kinase+activity+and+gene+expression+by+reactive+oxygen+species+and+their+role+in+vascular+physiology+and+pathophysiology.">Modulation of protein kinase activity and gene expression by reactive oxygen species and their role in vascular physiology and pathophysiology.</a> Arterioscler Thromb Vasc Biol. 20, 2175-2183 (2000).</li><li>Loh, K., Deng, H., Fukushima, A., Cai, X., Boivin, B., Galic, S., Bruce, C., Shields, B. J., Skiba, B., Ooms, L. M., Stepto, N., Wu, B., Mitchell, C. A., Tonks, N. K., Watt, M. J., Febbraio, M. A., Crack, P. J., Andrikopoulos, S., Tiganis, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reactive+oxygen+species+enhance+insulin+sensitivity.">Reactive oxygen species enhance insulin sensitivity.</a> Cell Metab. 10, 260-272 (2009).</li><li>Goldstein, B. J., Mahadev, K., Wu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Redox+paradox:+insulin+action+is+facilitated+by+insulin-stimulated+reactive+oxygen+species+with+multiple+potential+signaling+targets.">Redox paradox: insulin action is facilitated by insulin-stimulated reactive oxygen species with multiple potential signaling targets.</a> Diabetes. 54, 311-321 (2005).</li></ol>

一些病理情况，如炎症性疾病，癌症，缺血/再灌注，如放疗或化疗治疗（即顺铂）诱导ROS的生产过剩。因此，检测和测量ROS水平是很重要的，在很多的基础，临床前和临床研究。但是，活性氧有很短的半衰期，可能是复杂的检测。在这里，我们建议经常使用，并免费在哺乳动物细胞中的激进生产检测被广泛接受的简单测试。 使用染料的检测协议允许实验者使用各种广泛使用?...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> 5 - （和6） - 羧基-2&#39;，7&#39; -二氯荧光素二乙酸酯（羧基DCFDA） </td><td>分子探针</td><td> C369 </td><td>控制</td></tr><tr><td>羧基- H 2 DCFDA </td><td>分子探针</td><td> C400 </td><td></td></tr><tr><td>磺胺</td><td>西格玛</td><td> S9251 - 100G </td><td></td></tr><tr><td> N - 1 - napthylethylenediamine盐酸盐</td><td>西格玛</td><td> N9125 - 10G </td><td></td></tr><tr><td>亚硝酸盐标准</td><td>西格玛</td><td> 237213 - 100G </td><td></td></tr><tr><td> GSH / GSSG - GLO含量</td><td> Promega公司</td><td> V6612 </td><td>为了量化氧化，减少或氧化/还原型谷胱甘肽</td></tr></tbody>
</table>

production and detection of reactive oxygen species (ros) in cancers

活性氧分子，破坏DNA和RNA和氧化的蛋白质和脂质（脂质peroxydation）。这些反应的分子中含有氧和包括H 2 O 2（双氧水），NO（一氧化氮），O 2 - （氧化物阴离子），过氧亚硝基阴离子（ONOO - ），hydrochlorous酸（次氯酸）和羟自由基（OH - ） 。 氧化品种生产不仅病理情况下（癌症，缺血/再灌注，神经系统及心血管病症，传染性疾病，炎症性疾病，自身免疫性疾病等... ...），而且在非生理（病理）的情况下， 如细 ​​胞的新陈代谢3 4。事实上，活性氧在许多细胞信号通路中发挥重要作用（增殖，细胞活化5，6，7迁移等。） 。 ROS可以是有害的（然后被称为“氧化和nitrosative强调”）时，在细胞内的车厢和细胞产生高量一般回应ROS通过上调抗氧化剂，如超氧化物歧化酶（SOD）和过氧化氢酶（CAT），谷胱甘肽过氧化物酶（GPX）和谷胱甘肽（GSH），保护这些危险的自由基转换成无害的分子（即水）。维生素C和E也被形容为ROS的拾荒者（抗氧化剂）。 自由基是在3低量是有益的。巨噬细胞和中性粒细胞介导的免疫反应涉及NO的产生和释放，从而抑制病毒，病原体和肿瘤增殖8。 NO还与其他ROS反应，因此，也有作为解毒剂（活性氧清除剂）的作用。最后船只上的任何行为，调节血液流量，这是非常重要的肌肉适应长时间运动 9 ，10。一些出版物也表明，活性氧参与胰岛素SENSitivity 11，12。 许多方法来评价ROS产生。在这篇文章中，我们提出了几个简单，快速，经济实惠的的检测;这些实验已经验证了许多出版物，通常用来检测在哺乳动物细胞中ROS或其影响。虽然这些化验检测多种活性氧，其他检测只有一个单一的活性氧。

Watch this Scientific Journal Video about Production and Detection of Reactive Oxygen Species ROS in Cancers at JoVE.com

Production and Detection of Reactive Oxygen Species ROS in Cancers

在这里，我们提出简单的方法来测试和评估细胞活性氧的存在。

在癌症的生产和检测活性氧（ROS）的

Reactive oxygen species include a number of molecules that damage DNA and RNA and oxidize proteins and lipids (lipid peroxydation). These reactive ...

Research

JoVE Journal

Medicine

Production and Detection of Reactive Oxygen Species (ROS) in Cancers

Visualization and Analysis of Blood Flow and Oxygen Consumption in Hepatic Microcirculation: Application to an Acute Hepatitis Model

可视化和肝脏微循环血流量和耗氧量分析：急性肝炎模型中的应用

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of ...

科研

医学

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

癌症治疗监测的切伦科夫发光成像（CLI）

In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

Isolating and Using Sections of Bovine Mesenteric Artery and Vein as a Bioassay to Test for Vasoactivity in the Small Intestine

隔离和使用的牛肠系膜动脉和静脉部分，作为生物测定在小肠测试的Vasoactivity

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that ...

Assessing Transmissible Spongiform Encephalopathy Species Barriers with an In Vitro Prion Protein Conversion Assay

Assessing Transmissible Spongiform Encephalopathy Species Barriers with an In Vitro Prion Protein Conversion Assay

评估传染性海绵状脑病物种界限与<em&gt;在体外</em&gt;朊蛋白转化测定法

Studies to understanding interspecies transmission of transmissible spongiform encephalopathies (TSEs, prion diseases) are challenging in that they ...

Detection of Disease-associated &#945;-synuclein by Enhanced ELISA in the Brain of Transgenic Mice Overexpressing Human A53T Mutated &#945;-synuclein

Detection of Disease-associated ÃƒÅ½Ã‚Â±-synuclein by Enhanced ELISA in the Brain of Transgenic Mice Overexpressing Human A53T Mutated ÃƒÅ½Ã‚Â±-synuclein

发现疾病相关的α突触核蛋白通过增强ELISA在转基因小鼠表达人A53T的突变脑α突触核蛋白

In addition to established methods like Western blot, new methods are needed to quickly and easily quantify disease-associated &#945;-synuclein (&#945;SD) ...

Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury

Triggering Reactive Gliosis In Vivo by a Forebrain Stab Injury

触发反应胶质细胞增生<em&gt;在体内</em&gt;由前脑锐器伤

Following injury to the CNS, astrocytes undergo a broad range of biochemical, morphological, and molecular changes collectively referred to as reactive ...

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

体外和体内检测人类细胞中的 Mitophagy,线虫和小鼠

Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a ...

Analyzing Oxygen Consumption Rate in Primary Cultured Mouse Neonatal Cardiomyocytes Using an Extracellular Flux Analyzer

细胞外通量分析仪分析原代培养小鼠新生心肌细胞的耗氧率

Mitochondria and oxidative metabolism are critical for maintaining cardiac muscle function. Research has shown that mitochondrial dysfunction is an ...

Quantitative Mapping of Specific Ventilation in the Human Lung using Proton Magnetic Resonance Imaging and Oxygen as a Contrast Agent

以质子磁共振成像和氧为造影剂对人肺特异性通气进行定量定位

Specific ventilation imaging (SVI) is a functional magnetic resonance imaging technique capable of quantifying specific ventilation &#8213; the ratio of ...

Characterization of Sickling During Controlled Automated Deoxygenation with Oxygen Gradient Ektacytometry

受控自动脱氧过程中与氧梯度压质测定的病带特征

In sickle cell disease (SCD), a single point mutation in the gene coding for beta-globin causes the production of abnormal hemoglobin S (HbS). When ...