Monitoring the Protective Effects of Minocycline Treatment with Radiolabeled Annexin V in an Experimental Model of Focal Cerebral Ischemia
Xian Nan Tang 1,
Qing Wang 2,
Maya A. Koike 2,
Danye Cheng 3,
Michael L. Goris 3,
Francis G. Blankenberg 3,
and
Midori A. Yenari 2*
1 Department of Neurology, University of California, San Francisco, and San Francisco Veterans Affairs Medical Center, San Francisco, California; Department of Anesthesia, Stanford University, Stanford, California
2 Department of Neurology, University of California, San Francisco, and San Francisco Veterans Affairs Medical Center, San Francisco, California
3 Department of Radiology, Stanford University, Stanford, California
* To whom correspondence should be addressed. E-mail: yenari{at}alum.mit.edu.
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Abstract |
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Minocycline is an antibiotic now recognized to have antiapoptotic and antiinflammatory properties. Because of these properties, minocycline may be of benefit in reducing neuronal apoptosis from ischemia and subsequent postischemic inflammation if administered soon after a stroke. We now explore the feasibility of using 99mTc-annexin V, an in vivo marker of apoptosis, with SPECT to monitor the antiapoptotic effects of minocycline therapy. Methods: CB6/F1 adult male mice underwent unilateral distal middle cerebral artery occlusion (dMCA) occlusion and were imaged and sacrificed at 1, 3, 7, or 30 d after injury. Animals were given minocycline (or vehicle) 30 min and 12 h after dMCA occlusion and then given 22.5 mg/kg twice daily for up to 7 d. Before imaging, behavioral tests were performed to evaluate the neurologic function. After imaging, brains were collected for histology and assessed for the degree of apoptosis and microglial activation. Results: 99mTc-Annexin V uptake in injured hemispheres was significantly decreased 2- to 3-fold by minocycline at all time points. Minocyline reduced infarct size as seen histologically and improved behavioral indices as late as 30 d. Infarct volume as seen histologically correlated with radiolabeled annexin V uptake seen by SPECT. In situ fluorescent microscopy demonstrated that annexin V bound primarily to neurons at 1 and 3 d, with a shift toward microglia by 7 and 30 d. Conclusion: We found that minocycline significantly reduces neuronal apoptosis and infarct size and improves neurologic outcome in mice after acute focal cortical ischemia.
Key Words:
apoptosis, neuroprotective agents, in vivo imaging, stroke
