Ohnari Yoshito, Ueno Kazuhiro, Mori Kazuki, Kawashima Takayuki, Nishida Haruto, Higuchi Akihiro, Tokumaru Osamu, Miyamoto Shinji
Department of Cardiovascular Surgery, Oita University Faculty of Medicine, Oita, Japan.
Department of Cardiovascular Surgery, Oita University Faculty of Medicine, Oita, Japan.
Ann Vasc Surg. 2025 Mar;112:388-396. doi: 10.1016/j.avsg.2024.12.045. Epub 2024 Dec 26.
Acute ischemia in the hind extremities is a dangerous disease that causes irreversible damage. Revascularization procedures are important to prevent muscle damage, but these treatments may induce additional damage, also known as ischemia-reperfusion injury. The role of free radicals as pivotal mediators of ischemia-reperfusion injury remains a prominent hypothesis. We have recently revealed potent antioxidative activities of a novel free-radical scavenger named resorcimoline (RML). The present study aims to investigate RML as a new therapeutic agent to reduce muscle damage and prevent motor dysfunction of the hind extremities caused by acute limb ischemia.
Ischemia was induced in rats by occluding the femoral arteries in both hind limbs for 120 min with nylon bands, followed by reperfusion for 24 h. The RML group (n = 9) received an intravenous injection of RML immediately before reperfusion, whereas the saline group (n = 9) received an equivalent volume of saline. Motor function was evaluated by counting the number of steps required to return to normal gait. Serum biomarkers, including creatine kinase (CK) and lactate dehydrogenase (LDH), were measured to evaluate muscle injury. Muscle damage was assessed histologically with hematoxylin and eosin (HE) staining. Oxidative damage to DNA in muscle was evaluated by measuring the proportions of 8-hydroxy-2'-deoxyguanosine (8-OHdG)-positive cells by immunohistochemistry.
The average number of steps required to return to normal gait in the RML group was significantly smaller compared to the saline group (P = 0.04). Serum CK and LDH levels were significantly lower in the RML group than in the saline group (P = 0.03, P = 0.005). Histologically, the RML group demonstrated a significantly lower proportion of muscle damage (P = 0.004) and positivity of 8-OHdG (P = 0.01).
RML attenuated muscle damage and demonstrated protective effects against motor dysfunction after limb ischemia-reperfusion injury by reducing free-radical-induced DNA damage. RML can be a novel therapeutic agent that attenuates ischemia-reperfusion injury after acute limb ischemia.
后肢急性缺血是一种会导致不可逆损伤的危险疾病。血管再通手术对于预防肌肉损伤很重要，但这些治疗可能会引发额外损伤，即缺血再灌注损伤。自由基作为缺血再灌注损伤的关键介质，其作用仍然是一个重要的假说。我们最近发现了一种名为间苯二酚啉（RML）的新型自由基清除剂具有强大的抗氧化活性。本研究旨在探讨RML作为一种新的治疗药物，以减少肌肉损伤并预防急性肢体缺血所致后肢运动功能障碍。
通过用尼龙带结扎大鼠双后肢股动脉120分钟诱导缺血，随后再灌注24小时。RML组（n = 9）在再灌注前立即静脉注射RML，而生理盐水组（n = 9）注射等量的生理盐水。通过计算恢复正常步态所需的步数来评估运动功能。检测包括肌酸激酶（CK）和乳酸脱氢酶（LDH）在内的血清生物标志物，以评估肌肉损伤。用苏木精和伊红（HE）染色进行组织学评估肌肉损伤。通过免疫组化测量8-羟基-2'-脱氧鸟苷（8-OHdG）阳性细胞的比例，评估肌肉中DNA的氧化损伤。
与生理盐水组相比，RML组恢复正常步态所需的平均步数明显更少（P = 0.04）。RML组血清CK和LDH水平明显低于生理盐水组（P = 0.03，P = 0.005）。组织学上，RML组肌肉损伤比例和8-OHdG阳性率明显更低（P = 0.004，P = 0.01）。
RML减轻了肌肉损伤，并通过减少自由基诱导的DNA损伤，对肢体缺血再灌注损伤后的运动功能障碍表现出保护作用。RML可能是一种减轻急性肢体缺血后缺血再灌注损伤的新型治疗药物。
