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Thioredoxin Overexpression Enhances Neovascularization and Reduces Fibrosis In Mouse Hindlimb Ischemia Model
*Inam A Shaikh1,2, *Vaithinathan Selvaraju2, *Mahesh Thirunavukkarasu2, J Alexander Palesty1, *Nilanjana Maulik2
1Saint Mary's Hospital, Waterbury, CT;2University of Connecticut Health Center, Farmington, CT

Objective: Peripheral artery disease affects 12-20% Americans over the age of 60. Thioredoxin-1 (Trx-1) is a class of small redox proteins. We have demonstrated earlier that Trx-1 reduces oxidative stress resulting in less inflammation and increased angiogenesis in cardiac muscle via hemeoxygenase-1 and VEGF after MI. In the current study we evaluate the effect of Trx-1 on post-ischemic hindlimb recovery.
Design: NA
Setting: Basic science
Patients: NA
Interventions: Peripheral artery disease was mimicked using a hindlimb ischemia model. 8-12 week old wild type(WT) and thioredoxin-1 transgenic (Trx-1Tg/+) mice were subjected to femoral artery ligation. Following surgery, mice were observed for 5-weeks.
Main Outcome Measures: Serial laser doppler images were obtained and perfusion ratios between the ischemic and non-ischemic limbs were calculated at set time intervals. The ratios were compared between WT and Trx-1Tg/+ groups. Immunohistochemical analysis of the tissue was performed to quantify the extent of fibrosis, capillary and arteriolar density.
Results: The recovery of hind limb perfusion was significantly increased in Trx-1Tg/+ mice at day 7 [0.1709 ± 0.03 (n=10) vs. 0.3689 ± 0.07 (N=9); p<0.05], day-21 [0.4339 ± 0.08 (N=8) vs. 0.6240 ± 0.03 (N=9); p<0.05] and day 28[0.4194 ± 0.05 (N=8) vs. 0.9128 ± 0.11 (N=9); p<0.01]. Capillary density and arteriolar density staining showed significant increase in Trx-1Tg/+ mice (1265 ± 87 and 36 ± 2.4; counts/mm2 (n=5); p<0.05) as compared to WT (762 ± 86 and 22±1.09; counts/mm2 (n=5); p<0.05) mice. Picrosirrus Red and immunofluorescence staining showed decreased fibrosis and increased HO-1 expression respectively in Trx-1Tg/+ mice group as compared to WT mice.
Conclusions: Our results suggest that TRX-1 has therapeutic potential for peripheral artery disease and perhaps a possible cure.


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