Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.
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Method Article
Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure
In This Article
Summary
Abstract
Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol for bioluminescent optical imaging of HO-1-luciferase transgenic mice reported in this manuscript provides a rapid in vivo assay of the upregulation of the heme oxygenase-1 (HO-1) gene expression in response to the Gua Sha procedure. HO-1 has long been known to provide cytoprotection against oxidative stress. The upregulation of HO-1, assessed by the bioluminescence output, is thought to represent an antioxidative response to circulating hemoglobin products released by Gua Sha. Gua Sha was administered by repeated strokes of a smooth spoon edge over lubricated skin on the back or other targeted body part of the transgenic mouse until petechiae (splinter hemorrhages) or ecchymosis (bruises) indicative of extravasation of blood from subcutaneous capillaries was observed. After Gua Sha, bioluminescence imaging sessions were carried out daily for several days to follow the dynamics of HO-1 expression in multiple internal organs.
Protocol
- Female HO-1-luciferase transgenic mice (age of 4-6 weeks) can be purchased from Taconic Farms, Inc. Upon arrival, mice are placed in an animal housing facility for at least 4 days to allow accommodation.
- Preparation for bioluminescence imaging:
- Weigh each HO-1 mouse. Animal weight is required for calculating the dosage of luciferin.
- Hair removal: Use a cotton swap to apply hair remover Nair® over the abdomen and/or back of the animal. Waiting for 5 to 10 seconds. Use a clean cotton swap to wipe off hair. Dip a piece of tissue in distilled water to wipe clean remaining hair. ....
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Discussion
Transcription of HO-1, an inducible form of heme oxygenase, is upregulated by many factors including heme, hydrogen peroxide, UV irradiation, hypoxia, and physical stresses. Whole body imaging such as the reported HO-1 bioluminescence protocol provides a quick snapshot of systemic HO-1 expression in multiple organs. In small animals, bioluminescence imaging allows high-sensitivity in-vivo real-time quantitative longitudinal optical assessment of alterations in systemic gene expression, as shown. Bioluminescence molecul.......
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Disclosures
The authors have nothing to disclose.
Acknowledgements
K. Wong, S.T.C. Wong are supported by the funding from Functional and Molecular Imaging Center, Brigham and Women's hospital
K. Wong, S.T.C. Wong are supported by the Center for Biotechnology and Informatics, The Methodist Hospital Research Institute, Weill Cornell Medical College.
K.K. Kwong, I. Chen, J-Q Ren are supported by the funding from the Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital.
Lenuta Kloetzer, Braden Kuo are supported by funding from the Neuroenteric Research Center, Massachusetts General Hospital
The authors thank Drs. Q. Zeng and X. Xu of Optical Imaging Lab, Brigham and Women’s Ho....
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Materials
| Name | Company | Catalog Number | Comments |
|---|---|---|---|
| Luciferin | Caliper Life Sciences | P/N 122796 | Previously Xenogen |
| IVIS 100 | Caliper Life Sciences | 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 tr....
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