Distal Venous Arterialization Modulates Autophagy-Related Markers in Skeletal Muscle in a Rat Hindlimb Ischemia Model

Teguh Marfen Djajakusumah,1 Bethy Suryawathy Hernowo,2 Pei Ho,3 Herry Herman,4 Kiki Lukman,5 Ronny Lesmana6 1Division of Vascular and Endovascular Surgery, Department of Surgery, Universitas Padjadjaran – Hasan Sadikin Hospital, Bandung, Indonesia; 2Department of Pathology, Universitas Padjadjaran – Hasan Sadikin Hospital, Bandung, Indonesia; 3Department of Surgery, National University of Singapore, Singapore, Singapore; 4Department of Orthopaedic Surgery, Universitas Padjadjaran – Hasan Sadikin Hospital, Bandung, Indonesia; 5Department of Surgery, Universitas Padjadjaran – Hasan Sadikin Hospital, Bandung, Indonesia; 6Department of Physiology, Universitas Padjadjaran, Bandung, IndonesiaCorrespondence: Teguh Marfen Djajakusumah, Department of Vascular and Endovascular Surgery, Department of Surgery, Universitas Padjadjaran-Hasan Sadikin Hospital, Jl. Pasteur No. 38, Pasteur, Kec. Sukajadi, Bandung, Jawa Barat, 40161, Indonesia, Tel +628122023770, Email [email protected]: Distal venous arterialization (DVA) is a surgical option for limb salvage in peripheral arterial disease (PAD) without distal runoff. While increasingly applied, the skeletal muscle response after arterial–venous anastomosis remains unclear. This study evaluated skeletal muscle adaptation following DVA, focusing on autophagy regulation.Methods: This experimental preclinical study evaluated skeletal muscle adaptation following distal venous arterialization (DVA) in a rat hindlimb ischemia model. An experimental microsurgical study was conducted in 30 Wistar rats. Hindlimb ischemia (HLI) was induced by right femoral artery ligation. On day 7, rats were re-operated and divided into DVA, opened ligation (OL), or persistent ligation (HLI) groups, with contralateral limbs serving as sham. On day 14, skeletal muscles were harvested for capillary density (CD31) and Western blot analysis of HIF-1α, AMPK, LC3-II, and p62. Gait analysis (stride length and width) and limb thermography were also performed.Results: At day 7, HLI showed significant stride length reduction (p=0.029) and lower limb temperature (p< 0.001). By day 14, gait and thermography normalized in DVA and OL but remained impaired in HLI (p=0.041 and p< 0.001, respectively). Capillary density increased in HLI compared with sham (p=0.031), but not in DVA or OL. HIF-1α and AMPK were elevated in HLI (p=0.016 and p=0.011), while AMPK decreased in DVA (p=0.044). LC3-II expression was reduced in DVA compared with sham and HLI (p=0.002 and p=0.032). No significant differences were observed in p62.Conclusion: DVA enhances skeletal muscle recovery in ischemic limbs and is associated with normalization of hypoxia- and autophagy-related molecular markers.Keywords: autophagy, physiological adaptation, distal venous arterialization, hindlimb ischemia, LC3-II