Department of Surgery, Medical Faculty Manheim, Heidelberg University; European Center of Angioscience ECAS, Medical Faculty Manheim, Heidelberg University.
Computer-Assisted Clinical Medicine, Mannheim Institute for Intelligent Systems in Medicine, Medical Faculty Mannheim, Heidelberg University; Cooperative Core Facility Animal Scanner ZI, Medical Faculty Mannheim, Heidelberg University.
J Vis Exp. 2021 May 13(171). doi: 10.3791/62402.
The purpose of this study is to introduce and evaluate a modified surgical approach to induce acute ischemia in mice that can be implemented in most animal laboratories. Contrary to the conventional approach for double ligation of the femoral artery (DLFA), a smaller incision on the right inguinal region was made to expose the proximal femoral artery (FA) to perform DLFA. Then, using a 7-0 suture, the incision was dragged to the knee region to expose the distal FA. Magnetic resonance imaging (MRI) on bilateral hind limbs was used to detect FA occlusion after the surgery. At 0, 1, 3, 5, and 7 days after the surgery, functional recovery of the hind limbs was visually assessed and graded using the Tarlov scale. Histologic evaluation was performed after euthanizing the animals 7 days after DLFA. The procedures were successfully performed on the right leg in ten ApoE mice, and no mice died during subsequent observation. The incision sizes in all 10 mice were less than 5 mm (4.2 ± 0.63 mm). MRI results showed that FA blood flow in the ischemic side was clearly blocked. The Tarlov scale results demonstrated that hind limb function significantly decreased after the procedure and slowly recovered over the following 7 days. Histologic evaluation showed a significant inflammatory response on the ischemic side and reduced microvascular density in the ischemic hind limb. In conclusion, this study introduces a modified technique using a miniature incision to perform hind limb ischemia (HLI) using DLFA.
本研究旨在介绍和评估一种改良的手术方法，以诱导小鼠急性缺血，该方法可在大多数动物实验室实施。与传统的股动脉双重结扎（DLFA）方法不同，我们在右腹股沟区做一个较小的切口，以暴露股骨近端动脉（FA）进行 DLFA。然后，使用 7-0 缝线将切口拖至膝关节以暴露远端 FA。手术完成后，通过双侧后肢磁共振成像（MRI）检测 FA 闭塞。在手术后 0、1、3、5 和 7 天，使用 Tarlov 量表对后肢功能恢复进行视觉评估和分级。在 DLFA 后 7 天处死动物后进行组织学评估。在 10 只 ApoE 小鼠的右腿上成功完成了这些操作，且在后续观察过程中没有小鼠死亡。所有 10 只小鼠的切口均小于 5 毫米（4.2±0.63 毫米）。MRI 结果显示，缺血侧 FA 血流明显受阻。Tarlov 量表结果表明，手术后后肢功能明显下降，随后 7 天逐渐恢复。组织学评估显示缺血侧有明显的炎症反应，缺血后肢的微血管密度降低。总之，本研究介绍了一种改良技术，使用微型切口通过 DLFA 进行后肢缺血（HLI）。
