Targeted Ultrasound Imaging of Apoptosis with Annexin A5 Microbubbles in Acute Doxorubicin-Induced Cardiotoxicity
1
조회
0
다운로드
0
인용
0
참고문헌
초록
Background: The aim of this study was to assess the feasibility of targeted ultrasound imaging on apoptosis with annexin A5microbubbles (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.
Abstract
Background: The aim of this study was to assess the feasibility of targeted ultrasound imaging on apoptosis with annexin A5microbubbles (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.
참고문헌 (21)
- Bristow MR, "Clinical spectrum of anthracycline antibiotic cardiotoxicity", Cancer Treat Rep, 62, pp.873~879 (1978)
- Nousiainen T, "Early decline in left ventricular ejection fraction predicts doxorubicin cardiotoxicity in lymphoma patients", Br J Cancer, 86, pp.1697~1700 (2002)
- Saraste A, "Apoptosis in human acute myocardial infarction", Circulation, 95, pp.320~323 (1997)
- Olivetti G, "Apoptosis in the failing human heart", N Engl J Med, 336, pp.1131~1141 (1997)
- Saraste A, "Cardiomyocyte apoptosis and progression of heart failure to transplantation", Eur J Clin Invest, 29, pp.380~386 (1999)
- Nakamura T, "Fasmediated apoptosis in adriamycin-induced cardiomyopathy in rats: In vivo study", Circulation, 102, pp.572~578 (2000)
- Arola OJ, "Acute doxorubicin cardiotoxicity involves cardiomyocyte apoptosis", Cancer Res, 60, pp.1789~1792 (2000)
- Ling YH, "Apoptosis induced by anthracycline antibiotics in P388 parent and multidrug-resistant cells", Cancer Res, 53, pp.1845~1852 (1993)
- Zhang J, "Doxorubicin-induced apoptosis in spontaneously hypertensive rats: differential effects in heart, kidney and intestine, and inhibition by ICRF-187", J Mol Cell Cardiol, 28, pp.1931~1943 (1996)
- van Heerde WL, "Markers of apoptosis in cardiovascular tissues: focus on Annexin V", Cardiovasc Res, 45, pp.549~559 (2000)
- van Engeland M, "Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure", Cytometry, 31, pp.1~9 (1998)
- Kaufmann BA, "Molecular imaging with targeted contrast ultrasound", Curr Opin Biotechnol, 18, pp.11~16 (2007)
- Klibanov AL, "Targeted delivery of gas-filled microspheres, contrast agents for ultrasound imaging", Adv Drug Deliv Rev, 37, pp.139~157 (1999)
- Lindner JR, "Molecular imaging with contrast ultrasound and targeted microbubbles", J Nucl Cardiol, 11, pp.215~221 (2004)
- Bayne K, "Revised Guide for the Care and Use of Laboratory Animals available. American Physiological Society", Physiologist, 39, pp.199~ (1996)
- Jaffer FA, "Seeing within: molecular imaging of the cardiovascular system", Circ Res, 94, pp.433~445 (2004)
- Narula J, "Annexin-V imaging for noninvasive detection of cardiac allograft rejection", Nat Med, 7, pp.1347~1352 (2001)
- Hofstra L, "Visualisation of cell death in vivo in patients with acute myocardial infarction", Lancet, 356, pp.209~312 (2000)
- Dumont EA, "Real-time imaging of apoptotic cellmembrane changes at the single-cell level in the beating murine heart", Nat Med, 7, pp.1352~1355 (2001)
- Bennink RJ, "Annexin V imaging of acute doxorubicin cardiotoxicity (apoptosis) in rats", J Nucl Med, 45, pp.842~848 (2004)
- Wu S, "Adriamycin-induced cardiomyocyte and endothelial cell apoptosis: in vitro and in vivo studies", J Mol Cell Cardiol, 34, pp.1595~1607 (2002)