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Figure 4 The Extent of Apoptosis in Various Animal Groups Represented by Quantitative Annexin A5 Uptake and Histologic Characterization of Untreated and Broad Caspase-Inhibited Animals
The left panel demonstrates the uptake of annexin A5 in abdominal aortas. Disease control animals (C) with no atherosclerotic lesions are represented by an open bar and the remaining atherosclerotic animals by solid bars. Atherosclerotic animals imaged with mutant annexin were used as tracer controls (gray bar), and the remaining atherosclerotic animals were either untreated (black bar) or treated with various caspase inhibitors (colored bars). Broad and caspase-3 inhibition bars are shown in pink, caspase-8 and -9 inhibitors differentiating between mitochondrial and extramitochondrial pathways are in blue, and capase-1–specific inhibition is represented by a green solid bar. The statistical significance of caspase inhibition, in comparison with the disease control and untreated atherosclerotic animals, is shown above and below the bars, respectively. The uptake is significantly higher in the abdominal aortas of untreated atherosclerotic animals compared with that in the disease and tracer control groups. Treatment with a broad caspase inhibitor or selective caspase-1, -9, or -3 inhibitor significantly reduced the apoptotic activity as represented by lower annexin uptake. On the other hand, selective caspase-8 inhibition did not affect apoptosis. The right panel compares histologic characterization of atherosclerotic lesions in atherosclerotic untreated animals and atherosclerotic animals receiving the broad caspase inhibitor. Movat pentachrome (M5Ch), smooth muscle cell ( -actin), and macrophage (Ram-11) staining (x200) in untreated atherosclerotic animals (left) and broad caspase inhibitor-treated animals (right) demonstrate similar cholesterol crystal-rich necrotic core, foam cell-rich, and smooth muscle-deficient lesions. The morphologic characteristics of the lesions in untreated and caspase-treated atherosclerosis are unchanged. However, terminal deoxyribonucleotide transferase-mediated nick-end labeling (TUNEL) reveals evidence of marked apoptotic nuclei in untreated atherosclerotic animals but marked resolution in the atherosclerotic lesions in caspase-inhibited animals.
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