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目的研究在非胰岛素依赖型糖尿病状态下,小鼠肢体对缺血刺激的血管生成反应,并探讨其发生机制。方法雄性 C57BL/6小鼠40只,分别以常规饲料(对照组20只)和高脂肪饲料(糖尿病组20只)喂养。12周后,创建小鼠下肢动脉缺血模型,应用激光多普勒扫描仪记录肢体血流恢复情况。术后4周处死小鼠,取下肢肌肉组织行组织病理学检查,CD31染色,评价血管密度;取正常下肢肌肉组织分别行 ELISA 和 Western Blot 检测血管内皮生长因子(VEGF)及其下游因子 AKT 的表达状况,行逆转录聚合酶链反应检测 VEGF 受体的表达情况。结果非胰岛素依赖型糖尿病鼠肢体缺血后其自身恢复和新生血管密度明显低于正常对照组小鼠;肌肉组织中 VEGF 浓度明显高于对照组,而 VEGF 受体表达差异却无统计学意义。肌肉组织中 PAKT/AKT 比值明显低于对照组。结论非胰岛素依赖型糖尿病肢体对缺血刺激的反应明显减弱,这一现象可能与 VEGF 信号传导通路功能不全有关。
Objective To investigate the angiogenic response to ischemia-stimulated limbs in mice with non-insulin-dependent diabetes mellitus (DM) and to explore its mechanism. Methods Forty male C57BL / 6 mice were fed with normal diet (20 in control group) and high fat diet (20 in diabetic group). After 12 weeks, the model of lower extremity arterial ischemia in mice was established. The blood flow recovery of limbs was recorded by laser Doppler scanner. The mice were sacrificed at 4 weeks after operation, and the muscle tissue of the lower extremities was taken for histopathological examination and CD31 staining to evaluate the vascular density. The normal lower extremity muscle tissues were taken for detection of vascular endothelial growth factor (VEGF) and its downstream factor AKT by ELISA and Western Blot The expression of VEGF receptor was detected by reverse transcriptase-polymerase chain reaction. Results After ischemia in non-insulin-dependent diabetic mice, the self-recovery and neovascular density of the limbs were significantly lower than that of the normal control mice. The VEGF concentration in the muscle tissue was significantly higher than that in the control group, but the VEGF receptor expression was not statistically significant. The ratio of PAKT / AKT in muscle tissue was significantly lower than that in control group. Conclusion The response of non-insulin-dependent diabetic limbs to ischemic stimulation is obviously weakened. This phenomenon may be related to the dysfunction of VEGF signaling pathway.