Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

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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.

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Hypoxia-inducible factor-1 mediates transcriptional activation of the heme oxygenase-1 gene in response to hypoxia.

G Semenza, John Alam, Helen B Chin … (1997)

Exposure of rats to hypoxia (7% O2) markedly increased the level of heme oxygenase-1 (HO-1) mRNA in several tissues. Accumulation of HO-1 transcripts was also observed after exposure of rat aortic vascular smooth muscle (VSM) cells to 1% O2, and this induction was dependent on gene transcription. Activation of the mouse HO-1 gene by all agents thus far tested is mediated by two 5'-enhancer sequences, SX2 and AB1, but neither fragment was responsive to hypoxia in VSM cells. Hypoxia-dependent induction of the chloramphenicol acetyltransferase (CAT) reporter gene was mediated by a 163-bp fragment located approximately 9.5 kilobases upstream of the transcription start site. This fragment contains two potential binding sites for hypoxia-inducible factor 1 (HIF-1). A role for HIF-1 in HO-1 gene regulation was established by the following observations: 1) HIF-1 specifically bound to an oligonucleotide spanning these sequences, 2) mutation of these sequences abolished HIF-1 binding and hypoxia-dependent gene activation in VSM cells, 3) hypoxia increased HIF-1alpha and HIF-1beta protein levels in VSM cells, and 4) hypoxia-dependent HO-1 mRNA accumulation was not observed in mutant hepatoma cells lacking HIF-1 DNA-binding activity. Taken together, these data demonstrate that hypoxia induces HO-1 expression in animal tissues and cell cultures and implicate HIF-1 in this response.

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Visualizing gene expression in living mammals using a bioluminescent reporter.

B Frank Eames, D Benaron, Pamela Contag … (1997)

Control of gene expression often involves an interwoven set of regulatory processes. As information regarding regulatory pathways may be lost in ex vivo analyses, we used bioluminescence to monitor gene expression in living mammals. Viral promoters fused to firefly luciferase as transgenes in mice allowed external monitoring of gene expression both superficially and in deep tissues. In vivo bioluminescence was detectable using either intensified or cooled charge-coupled device cameras, and could be detected following both topical and systemic delivery of substrate. In vivo control of the promoter from the human immunodeficiency virus was demonstrated. As a model for DNA-based therapies and vaccines, in vivo transfection of a luciferase expression vector (SV-40 promoter and enhancer controlling expression) was detected. We conclude that gene regulation, DNA delivery and expression can now be noninvasively monitored in living mammals using a luciferase reporter. Thus, real-time, noninvasive study of gene expression in living animal models for human development and disease is possible.

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Bioluminescent indicators in living mammals.

Claire Stevenson, Pamela Contag, C Contag … (1998)

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Author and article information

Journal

Journal ID (nlm-ta): J Vis Exp

Journal ID (publisher-id): JoVE

Title: Journal of Visualized Experiments : JoVE

Publisher: MyJove Corporation

ISSN (Electronic): 1940-087X

Publication date Collection: 2009

Publication date (Electronic): 28 August 2009

Issue: 30

Electronic Location Identifier: 1385

Affiliations

Department of Radiology, Havard Medical School, Massachusetts General Hospital

Athinoula A. Martinos Center for Biomedical Imaging, Havard Medical School, Massachusetts General Hospital

Gastrointestinal Unit, Havard Medical School, Massachusetts General Hospital

Department of Medicine, Havard Medical School, Massachusetts General Hospital

Center for biotechnology and Informatics, The Methodist Hospital Research Institute

Department of Radiology, Weill Cornell Medical College of Cornell University

Bejing University of Chinese Medicine

Department of Health Technology and Informatics, The Hong Kong Polytechnic University - PolyU

Department of Radiology, Harvard Medical School

Author notes

Correspondence to: Kenneth K. Kwong at kwong@ 123456nmr.mgh.harvard.edu

Article

Publisher ID: 1385

DOI: 10.3791/1385

PMC ID: 3149908

PubMed ID: 19718012

SO-VID: 686153dd-bd2e-4c2f-9614-ca65be566c71

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This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

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Abstract

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Electron Channelling Contrast Imaging (ECCI)

Most cited references 18

Hypoxia-inducible factor-1 mediates transcriptional activation of the heme oxygenase-1 gene in response to hypoxia.

Visualizing gene expression in living mammals using a bioluminescent reporter.

Bioluminescent indicators in living mammals.

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