目的 探讨腰交感干切除术对大鼠下肢缺血模型中侧支血管形成及血管新生的促进作用。 方法 建立 Sprague-Dawley 大鼠股动脉结扎缺血模型，随机分为对照组和交感切除组，后者行双侧腰交感干切除术。术后第 14 天，通过 X 线造影及 Micro-CT 三维重建观察缺血肢体侧支血管形成情况，并测量管径。采用免疫荧光双标法，以 α-SMA 和 CD31 标记血管结构，同时使用 Ki67 染色评估血管平滑肌细胞增殖活性，从多角度分析血管修复与再生。术后第 3、7及14 天，利用激光散斑成像检测足底血流灌注恢复情况；术后第 30 天，行 H&E 染色测量腓肠肌纤维横截面积，统计 CD31 阳性微血管数量，并检测术后3天 VEGFA 蛋白表达水平；术后第 7 天，通过流式细胞术分析患肢内收肌群中巨噬细胞 M1/M2 极化比例。 结果 与对照组相比，交感切除组患肢侧支血管平均管径显著增大（0.52±0.09 mm vs. 0.38±0.07 mm，P<0.01），α-SMA⁺/Ki67⁺ 细胞数量明显增加（5.38±0.87 vs. 2.22±0.64，P<0.01）。术后第 3、7及14 天，交感切除组足底血流灌注恢复率均显著升高（P<0.05）。术后第 30 天，腓肠肌纤维横截面积明显增大（1706±92 μm² vs. 1145±132 μm²，P<0.001），CD31 阳性微血管数量增加（171±19 vs. 93±8，P<0.001），VEGFA 蛋白表达显著上调（P<0.01）。此外，交感切除组 M2 型巨噬细胞比例显著升高[（28.7±3.0）% vs. （17.8±2.2）%，P<0.01]，而 M1 型比例无显著差异。 结论 腰交感干切除术可有效促进大鼠缺血下肢的侧支血管生长与血管新生，其作用机制可能与诱导巨噬细胞向 M2 型极化、上调 VEGFA 表达相关，为临床治疗缺血性下肢疾病提供了潜在的干预思路。
Abstract
Objective To investigate the effect of lumbar sympathectomy on collateral vessel formation and angiogenesis in a rat model of hindlimb ischemia. Methods A femoral artery ligation-induced hindlimb ischemia model was established in Sprague-Dawley rats, which were randomly divided into a control group and a sympathectomy group. The latter underwent bilateral lumbar sympathectomy. On postoperative day 14, collateral vessel formation in the ischemic limb was assessed by X-ray angiography and Micro-CT three-dimensional reconstruction, and vessel diameters were measured. Immunofluorescence double staining with α-SMA and CD31 was performed to visualize vascular structures, and Ki67 staining was used to evaluate the proliferative activity of vascular smooth muscle cells, thereby comprehensively analyzing vascular repair and regeneration. Blood perfusion in the plantar region was assessed on postoperative days 3, 7, and 14 using laser speckle imaging. On day 30, hematoxylin and eosin (H&E) staining was used to measure gastrocnemius fiber cross-sectional area, count CD31-positive microvessels, and detect VEGFA protein expression (day 3). On day 7, flow cytometry was performed to analyze the polarization ratio of M1/M2 macrophages in the adductor muscle group. Results Compared with the control group, the sympathectomy group exhibited a significantly larger mean collateral vessel diameter (0.52±0.09 mm vs. 0.38±0.07 mm, P<0.01) and increased numbers of α-SMA⁺/Ki67⁺ cells (5.38±0.87 vs. 2.22±0.64, P<0.01). Blood perfusion recovery rates on postoperative days 3, 7, and 14 were significantly improved (P<0.05). By day 30, gastrocnemius fiber cross-sectional area was markedly enlarged (1706±92 μm² vs. 1145±132 μm², P<0.001), CD31-positive microvessel counts were increased (171±19 vs. 93±8, P<0.001), and VEGFA expression was significantly upregulated (P<0.01). Moreover, the proportion of M2 macrophages was significantly elevated (28.7±3.0% vs. 17.8±2.2%, P<0.01), while the proportion of M1 macrophages showed no significant difference. Conclusions Lumbar sympathectomy effectively promotes collateral vessel growth and angiogenesis in ischemic rat hindlimbs. The underlying mechanism may involve the induction of macrophage polarization toward the M2 phenotype and upregulation of VEGFA expression.
关键词
腰交感干切除术 /
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下肢缺血 /
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侧支血管生长 /
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血管新生 /
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巨噬细胞极化
Key words
Lumbar sympathectomy /
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Hindlimb ischemia /
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Collateral vessel growth /
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Angiogenesis /
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Macrophage polarization
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