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Akt, p70 S6-kinase and Bcl-2 protein expression in EPCs from young (Y) and older (O) men. Protein abundance is expressed relative to β-actin to account for variations in protein loading and is shown normalized to the young mean. Values are means SEM. * p
The mechanisms responsible for the apparent proapoptotic EPC phenotype with aging are unclear. Factors such as inflammation and oxidative stress that are known to increase with age may play a role [31,32]. Indeed, the pro-inflammatory cytokine C-reactive protein, that generally increases with age, has been shown to induce EPC apoptosis in vitro . To elucidate possible signaling mechanisms that may be responsible for the greater apoptotic susceptibility of EPCs in older adults, we determined the relative expression of several upstream proteins in the PI-3-kinase/Akt signaling pathway known to effect caspase-3 activation. This pathway plays a vital role in cell survival by initiating downstream activation of antiapoptotic proteins and suppression of cell death signals . Perturbations in PI-3-kinase/Akt signaling may lead to either excessive cell death or a number of disease processes, such as in cancer [34,35]. In the present study, we demonstrated that aging is associated with decreased expression of antiapoptotic proteins involved in the PI-3-kinase/Akt signaling pathway. Specifically, older men had lower Akt, p70 S6-kinase and Bcl-2 expression compared with young men. Previous studies have shown that constitutive activation of Akt and p70 S6-kinase promotes cell survival, while pharmacological inhibition of these proteins increases apoptosis . Similarly, suppression of Bcl-2 expression results in greater cell death in multiple cell types [37,38]. Thus, the greater apoptotic susceptibility in EPCs from older men may be due, at least in part, to overall lower expression of antiapoptotic proteins associated with the PI-3-kinase/Akt signaling pathway. In addition, telomerase activity has been shown to be inversely associated with intracellular caspase-3 activation . Considering we observed approximately 60% less telomerase activity in EPCs from older men compared with younger men, it is possible that age-related reductions in telomerase activity may contribute to enhanced caspase-3 activation.
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