Effects of aging and sex on redox-regulation of ischemic vascularization

• DOI: 10.1016/j.freeradbiomed.2017.10.068
• Publication Date: 2017-11-20T19:03:36Z
• Authors: Beatriz Ferran Perez; Yuko Tsukahara; Yoshimitsu Yura; Markus M. Bachschmid; Reiko Matsui
• Keywords: 0301 basic medicine; 03 medical and health sciences; 3. Good health

Glutathione (GSH) adducts formation on protein thiols (S-glutathionylation), an oxidative post-translational modification, is reversed by glutaredoxin-1 (Glrx). We previously found that Glrx transgenic mice showed impaired blood flow recovery after femoral artery ligation (Murdoch 2014), while this ischemic revascularization was improved in Glrx knockout mice in association with the hypoxia-inducible factor-1α activation (Watanabe 2016). Ischemic limb recovery is blunted by aging. Also, clinical studies indicate that ischemic limb recovery is worse in female than male patients. Mouse studies showed poor recovery of hindlimb ischemia in females. Although it is considered that Glrx activity is decreased in aged animals, the regulation of Glrx, anti-angiogenic enzyme, by aging or sex is not well studied. Using mouse (4-24 months old) tissues obtained from NIA tissue bank, we assessed overall GSH adducts in muscles by Western blot with an anti-GSH antibody. Glrx activity in skeletal muscles and serum of male and female mice was measured using a Glutaredoxin Assay Kit (IMCO) that uses fluorescent eosin-GSSG as a substrate, more sensitive than usual assay measuring NADPH consumption. Our data showed an increase of GSH adducts in skeletal muscle of male and female mice of 12 and 24 months old, comparing with 4 months old mice. However, surprisingly muscle tissues from older mice (12 and 24 months old) showed a 2-fold increase in Glrx activity than muscles of younger mice [males and females: p