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目的:探讨疏肝活血方对肢体缺血后代偿性血管新生及相关基因表达的调节作用。方法:BALB/c裸鼠120只,随机分为疏肝活血方组、疏肝方组、活血方组、模型对照组和正常对照组,股动脉结扎法建立裸鼠肢体缺血模型,分别于术后1、2、3、4周观察各组裸鼠肢体缺血状态的改变,应用苏木素-伊红(HE)染色和CD34免疫组化染色观察肌肉组织一般形态学及微血管数的改变,应用Western蛋白质印迹检测低氧诱导因子-1α(HIF-1α)、肝细胞生长因子(HGF)和血管内皮生长因子(VEGF)基因在肌肉组织中的表达。结果:与模型对照组相比,疏肝活血方能明显改善裸鼠肢体的缺血状态。术后2-4周,疏肝活血方组裸鼠缺血肢体肌肉中的微血管数(MVC)明显高于模型对照组(P<0.01),缺血肢体肌肉中HIF-1α、HGF和VEGF的蛋白质表达明显强于其它各组(P<0.05)。结论:疏肝活血方具有明显的促血管生成作用,能够明显上调血管新生相关基因HIF-1α、HGF和VEGF的蛋白质表达水平。
Objective: To investigate the regulation effect of Shugan Huoxue Recipe on compensatory angiogenesis and related gene expression after limb ischemia. Methods: One hundred and twenty BALB / c nude mice were randomly divided into three groups: Shugan Huoxue recipe group, Shuganfang recipe group, Huoxue recipe group, model control group and normal control group. Limb ischemia model was established in nude mice by ligation of femoral artery. At 1, 2, 3 and 4 weeks after operation, the changes of ischemic limbs of nude mice in each group were observed. HE staining and CD34 immunohistochemistry were used to observe the changes of muscle morphology and microvessel number. Western blot was used to detect the expression of HIF-1α, HGF and VEGF in muscle tissue. Results: Compared with the model control group, Shugan Huoxue Decoction can significantly improve the ischemic status of nude mice. At 2-4 weeks after operation, the number of MVC in SHG group was significantly higher than that in model control group (P <0.01), the expression of HIF-1α, HGF and VEGF in ischemic limb muscle Protein expression was significantly stronger than other groups (P <0.05). Conclusion: Shugan Huoxue Recipe has obvious angiogenic effect, and can significantly upregulate the protein expression of angiogenesis-related genes HIF-1α, HGF and VEGF.