Annexin V imaging detects diabetes-accelerated apoptosis and monitors the efficacy of benfotiamine treatment in ischemic limbs of mice.
The role of apoptosis imaging for monitoring treatment response in ischemic limbs has not been properly explored. In this study, we investigated the ability of annexin V (AnxV) imaging to assess the efficacy of antiapoptotic treatment in ischemic limbs of diabetic mice. Normal C57BL/6 mice and streptozotocin-induced diabetic mice were subject to hindlimb ischemia. AnxV-conjugated fluorescent streptavidin probes were intravenously injected, and optical imaging was performed. Tissue apoptosis was quantified by histochemistry and Western blotting. The AnxV probes showed specific targeting to apoptotic cells on confocal microscopy and flow cytometry. Intravenous AnxV probes displayed substantially greater accumulation in ischemic limbs of diabetic mice. Benfotiamine (BFT) treatment of diabetic mice led to better perfusion recovery on laser Doppler imaging and reduced AnxV binding on optical imaging. TUNEL staining and cleaved caspase-3 Western blots confirmed accelerated apoptosis by diabetes and its suppression by BFT treatment. Furthermore, AnxV-SAv-PEcy5.5 uptake in the ischemic limbs closely correlated to cleaved caspase-3 expression. Thus, AnxV imaging may be useful for monitoring the efficacy of therapeutic agents designed to suppress ischemia-induced apoptosis.
Kyung-Ho
KH
Jung
Jin Hee
JH
Lee
Jin Won
JW
Park
Jin Young
JY
Paik
Cung Hoa Thien
CH
Quach
Eun Jeong
EJ
Lee
Kyung-Han
KH
Lee
D000818:Animals; D017304:Annexin A5 / Q000008:administration & dosage / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D003921:Diabetes Mellitus, Experimental / Q000150:complications / Q000188:drug therapy / Q000473:pathology; D006614:Hindlimb / Q000098:blood supply / Q000473:pathology*; D007511:Ischemia / Q000188:drug therapy* / Q000473:pathology*; D017078:Laser-Doppler Flowmetry; D051379:Mice; D008810:Mice, Inbred C57BL; D019809:Streptavidin / Q000008:administration & dosage / Q000494:pharmacology; D013831:Thiamine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000627:therapeutic use
D017304:Annexin A5; D019809:Streptavidin; D013831:Thiamine; C013835:benphothiamine