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Targeted ultrasound imaging of apoptosis with annexin a5 microbubbles in acute Doxorubicin-induced cardiotoxicity.

 Pil-Ki Min  ;  Soyeon Lim  ;  Soo-Jung Kang  ;  Sung-Yu Hong  ;  Ki-Chul Hwang  ;  Kwang-Hoe Chung  ;  Chi Young Shim  ;  Se-Joong Rim  ;  Namsik Chung 
 Journal of Cardiovascular Ultrasound, Vol.18(3) : 91-97, 2010 
Journal Title
Journal of Cardiovascular Ultrasound
Issue Date
Adult ; Animals ; Annexin A5 ; Aorta ; Apoptosis ; Avidin ; Cardiomyopathies ; Doxorubicin ; Echocardiography ; Flow Cytometry ; Fluorescein-5-isothiocyanate ; Fluorescence ; Heart ; Humans ; Hydrogen Peroxide ; In Situ Nick-End Labeling ; Male ; Microbubbles ; Myocardium ; Myocytes, Smooth Muscle ; Rats
AnnexinA5 ; Apoptosis ; Cardiomyopathy ; Echocardiography ; Microbubbles
BACKGROUND: The aim of this study was to assess the feasibility of targeted ultrasound imaging on apoptosis with annexin A5 microbubbles (A5MB) in acute doxorubicin-induced cardiotoxicity.

METHODS: Avidinated and octafluoropropan-filled phospholipid microbubbles were conjugated with biotinylated annexin A5. To confirm the specific binding of A5MB, flow cytometry was performed with hydrogen peroxide induced apoptosis in rat aorta smooth muscle cells incubated with fluorescein-5-isothiocyanate (FITC) labeled annexin A5 and A5MB. Adult male rats were injected intraperitoneally with 5 mg/kg doxorubicin weekly for 3 weeks (n = 5). Control rats were injected with normal saline (n = 5). At 24 hours after the final treatment, triggering imaging was performed 15 min after an intravenous bolus injection of A5MB for washout of freely circulating microbubbles. After echocardiography, the heart was isolated for histological detection of apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay.

RESULTS: In the in vitro tests, fluorescence intensity was low for healthy cells and high for apoptotic cells when incubated with FITC-labeled annexin A5 and A5MB. Rats treated with doxorubicin showed significant contrast opacification of the myocardium on contrast echocardiography using A5MB. However, no opacification was observed in control rats. Apoptosis was confirmed by TUNEL assay in doxorubicin treated rats.

CONCLUSION: Acute doxorubicin-induced cardiomyopathy based on early apoptosis can be assessed and imaged with targeted ultrasound imaging using A5MB in rats.
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1. College of Medicine (의과대학) > Research Institute (부설연구소) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
Yonsei Authors
Min, Pil Ki(민필기) ORCID logo https://orcid.org/0000-0001-7033-7651
Shim, Chi Young(심지영) ORCID logo https://orcid.org/0000-0002-6136-0136
Rim, Se Joong(임세중) ORCID logo https://orcid.org/0000-0002-7631-5581
Lim, So Yeon(임소연)
Chung, Nam Sik(정남식)
Hwang, Ki Chul(황기철)
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