血红素加氧酶1表达上调在刮痧过程中的生物发光成像
1Department of Radiology,Massachusetts General Hospital, Harvard Medical School, 2Athinoula A. Martinos Center for Biomedical Imaging,Massachusetts General Hospital, Harvard Medical School, 3Gastrointestinal Unit,Massachusetts General Hospital, Harvard Medical School, 4Department of Medicine,Massachusetts General Hospital, Harvard Medical School, 5Center for biotechnology and Informatics,The Methodist Hospital Research Institute, 6Department of Radiology,The Methodist Hospital, Weill Cornell Medical College, 7Bejing University of Chinese Medicine, 8Department of Health Technology and Informatics,The Hong Kong Polytechnic University, 9Department of Radiology,Brigham and Women’s Hospital, Harvard Medical School
Summary
中国传统的治疗皮肤刮，刮痧，导致皮下微血管的血液外渗。我们报告一个HO – 1的生物发光成像的协议<em>荧光素酶</em转基因小鼠证明，刮痧上调血红素氧合- 1（HO – 1）在多器官。
Abstract
刮痧是一种传统的中国民间疗法，采用皮肤刮引起皮下微血管的血液外渗和瘀伤。发光的光学成像协议HO – 1 -<em>荧光素酶</em报告>在这个手稿的转基因小鼠提供了一个快速<em>在体内</em>检测的上调血红素氧合酶-1（HO – 1）在刮痧过程中的基因表达。 HO – 1一直提供抗氧化应激的细胞保护作用。上调HO – 1，生物发光输出评估，被认为是代表循环血红蛋白刮痧发布的产品具有抗氧化反应。刮痧是管理比润滑皮肤瘀斑（分裂出血）或瘀斑（跌打损伤）观察表明从皮下毛细血管的血液外渗，直到背面或其他有针对性的转基因小鼠的身体的一部分的顺利勺边重复笔画。刮痧后，进行了生物发光成像会几天每天跟随多个内脏器官的HO – 1表达的动态。
Protocol
女HO – 1的荧光素酶的转基因小鼠（年龄4-6周）可从Taconic的农场，公司购买的，到达目的地后，小鼠被放置至少4天，让住宿在畜舍设施。 生物发光成像的准备工作： 称量每个HO – 1的鼠标。动物的体重是需要计算的荧光素的用量。 脱毛：用棉花交换申请超过头发卸妆奈尔®腹部和/或背面的动物。等待5到10秒。使用干净的棉布擦去头发的交换。?…
Discussion
，血红素加氧酶的诱导形成，HO – 1的转录上调的因素很多，包括血红素，过氧化氢，紫外线照射，缺氧，和物理应力。如全身显像报道，HO – 1生物发光协议提供了一个全身多个器官中的表达，HO – 1的快照。在小动物，生物发光成像允许高灵敏度体内实时定量全身基因表达的改变纵向光学评估，如图所示。生物荧光分子成像，降低了成本，增加了小动物的基因表达分析的吞吐量，使得实际，迅速实…
Divulgations
The authors have nothing to disclose.
Acknowledgements
K.黄，黄STC是从功能和分子影像中心，Brigham和妇女医院的经费支持
K.黄，黄STC支持生物技术和信息中心，循道卫理医院研究所，康奈尔大学威尔医学院。
KK广，JQ任一陈Athinoula答：Martinos，马萨诸塞州总医院的生物医学成像中心的资金支持。
Lenuta Kloetzer，布雷登郭从Neuroenteric研究中心，美国马萨诸塞州总医院的资金支持
作者感谢博士。问：曾和X旭光学成像实验室，布里格姆妇女医院的技术帮助。
Materials
| Material Name | Type | Company | Catalogue Number | Comment |
|---|---|---|---|---|
| Luciferin | Caliper | P/N 122796 | Previously Xenogen | |
| IVIS 100 | Caliper | Previously Xenogen |
References
- Zhang, W. Selection of potential therapeutics based on in vivo spatiotemporal transcription patterns of heme oxygenase-1. J Mol Med. 80, 655-6564 (2002).
- Zhang, W. Rapid in vivo functional analysis of transgenes in mice using whole body imaging of luciferase expression. Transgenic Res. 10, 423-434 (2001).
- Contag, C. H., Stevenson, D. K. In vivo patterns of heme oxygenase-1 transcription. J Perinatol. 21, 119-1124 (2001).
- Nielsen, A., Knoblauch, N. T., Dobos, G. J., Michalsen, A., Kaptchuk, T. J. The effect of Gua Sha treatment on the microcirculation of surface tissue: a pilot study in healthy subjects. Explore (NY). 3, 456-466 (2007).
- Schwickert, M. E., Saha, F. J., Braun, M., Dobos, G. J. Gua Sha for migraine in inpatient withdrawal therapy of headache due to medication overuse. Forsch Komplementmed. 14, 297-300 (2007).
- Tsai, P. S., Lee, P. H., Wang, M. Y. Demographics, training, and practice patterns of practitioners of folk medicine in Taiwan: a survey of the Taipei metropolitan area. J Altern Complement Med. 14, 1243-1248 (2008).
- Nielsen, A. Gua sha research and the language of integrative medicine. J Bodyw Mov Ther. 13, 63-72 (2009).
- Contag, C. H. Visualizing gene expression in living mammals using a bioluminescent reporter. Photochem Photobiol. 66, 523-531 (1997).
- Cui, K., Xu, X., Zhao, H., Wong, S. T. A quantitative study of factors affecting in vivo bioluminescence imaging. Luminescence. 23, 292-295 (2008).
- Shibahara, S., Yoshida, T., Kikuchi, G. Mechanism of increase of heme oxygenase activity induced by hemin in cultured pig alveolar macrophages. Arch Biochem Biophys. 197, 607-617 (1979).
- Lee, P. J. Hypoxia-inducible factor-1 mediates transcriptional activation of the heme oxygenase-1 gene in response to hypoxia. J Biol Chem. 272, 5375-5381 (1997).
- Maines, M. D. The heme oxygenase system: a regulator of second messenger gases. Annu Rev Pharmacol Toxicol. 37, 517-554 (1997).
- Wunder, C., Potter, R. F. The heme oxygenase system: its role in liver inflammation. Curr Drug Targets Cardiovasc Haematol Disord. 3, 199-208 (2003).
- Hoekstra, K. A., Godin, D. V., Cheng, K. M. Protective role of heme oxygenase in the blood vessel wall during atherogenesis. Biochem Cell Biol. 82, 351-359 (2004).
- Yang, N. C., Lu, L. H., Kao, Y. H., Chau, L. Y. Heme oxygenase-1 attenuates interleukin-1beta-induced nitric oxide synthase expression in vascular smooth muscle cells. J Biomed Sci. 11, 799-809 (2004).
- Morse, D., Choi, A. M. Heme oxygenase-1: from bench to bedside. Am J Respir Crit Care Med. 172, 660-670 (2005).
- Ryter, S. W. Protective functions of heme oxygenase-1 and carbon monoxide in the respiratory system. Antioxid Redox Signal. 9, 2157-2173 (2007).
- Wilson, K., Yu, J., Lee, A., Wu, J. C. In vitro and in vivo bioluminescence reporter gene imaging of human embryonic stem cells. J Vis Exp. , (2008).
- Calabrese, V. Vitagenes, dietary antioxidants and neuroprotection in neurodegenerative diseases. Front Biosci. 14, 376-397 (2009).
- Contag, P. R., Olomu, I. N., Stevenson, D. K., Contag, C. H. Bioluminescent indicators in living mammals. Nat Med. 4, 245-247 (1998).
Citer Cet Article
Kwong, K. K., Kloetzer, L., Wong, K. K., Ren, J., Kuo, B., Jiang, Y., Chen, Y. I., Chan, S., Young, G. S., Wong, S. T. Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure. J. Vis. Exp. (30), e1385, doi:10.3791/1385 (2009).