Overexpression of Thioredoxin1 enhances functional recovery in mouse model of hind limb ischemia

• Published in: The Journal of surgical research 2017-08, Vol.216, p.158-168
• Main Authors: Shaikh, Inam A., MD, Rishi, Mohammad Tipu, MD, Youssef, Mark, MD, Selvaraju, Vaithinathan, PhD, Thirunavukkarasu, Mahesh, PhD, Ukani, Gopi, MD, Lakshmanan, Rajesh, PhD, Palesty, J. Alexander, MD, FACS, Maulik, Nilanjana, PhD, FAHA
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animals; Biomarkers - metabolism; Blood perfusion; Blotting, Western; Gene therapy; Genetic Therapy - methods; Hind limb ischemia; Hindlimb - blood supply; Hindlimb - metabolism; Immunohistochemistry; Ischemia - genetics; Ischemia - metabolism; Ischemia - therapy; Male; Mice; Mice, Inbred C57BL; Mice, Transgenic; Peripheral Vascular Diseases - genetics; Peripheral Vascular Diseases - metabolism; Peripheral Vascular Diseases - therapy; Surgery; Thioredoxin1; Thioredoxins - genetics; Thioredoxins - metabolism; Up-Regulation
• ISSN: 0022-4804; 1095-8673
• DOI: 10.1016/j.jss.2017.04.019

Abstract Background There is keen interest in finding nonsurgical treatments for peripheral vascular disease (PVD). Previously, we demonstrated that selective activation of Thioredoxin1 (Trx1), a 12-kDa cytosolic protein, initiates redox-dependent signaling and promotes neovascularization after ischemic heart disease. Therefore, Trx1 might possess immense potential to not only treat murine hind limb ischemia (HLI) through effective angiogenesis but also provide PVD patients with nonsurgical therapy to enhance neovascularization and improve blood perfusion. Methods To determine whether activation of Trx1 increases blood perfusion in HLI, two different strategies were used—gene therapy and transgenic model system. In adenoviral-mediated gene therapy, 8- to 12-wk-old mice were divided into two groups: (1) control Adeno-LacZ (Ad-LacZ) and (2) Adeno-Thiroedoxin1 (Ad-Trx1). The mice underwent surgical intervention to induce right HLI followed by injection with Ad-LacZ or Ad-Trx1, respectively. In the second strategy, we used wild-type and transgenic mice overexpressing Trx1 (Trx1Tg/+ ). All the animals underwent Doppler imaging for the assessment of limb perfusion followed by immunohistochemistry and Western blot analysis. Results Significant increases in perfusion ratio were observed in all the Trx1 overexpressed groups compared with their corresponding controls. Expressions of heme oxygenase-1, vascular endothelial growth factor, and the vascular endothelial growth factor receptors Flk-1 and Flt-1 were increased in Trx1 overexpressed mice compared with their respective controls. Blood perfusion in the ischemic limb gradually improved and significantly recovered in Trx1Tg/+ and Ad-Trx1 groups compared with their corresponding controls. The capillary and arteriolar density in the ischemic zone were found to be higher in Trx1Tg/+ group compared with wild type. Conclusions The overall outcomes of our study demonstrate that Trx1 enhances blood perfusion and increases angiogenic protein expression in a rodent HLI model. These results suggest that Trx1 is a potential target for clinical trials and drug therapy for the treatment of PVD.