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目的研究采用脐血内皮祖细胞局部注射治疗糖尿病下肢缺血有效性及机制,为临床治疗糖尿病足病提供理论依据。方法 1取产妇足月产脐血分离单核细胞,培养7天,行流式细胞仪鉴定细胞,并计数活细胞数。2用Wistar雄性大鼠分成三组:①糖尿病射线照射后结扎双后肢股动脉,局部注射EPC(DLM)。②糖尿病结扎双后肢股动脉,局部注射PBS(DLC)。③正常大鼠射线照射后结扎双后肢股动脉,局部注射EPC(NLM)。3用GFP(绿色荧光)示踪EPC,Ⅷ因子免疫组化检测肌纤维间毛细血管数,RT-PCR检测双后肢肌肉VEGFmRNA水平。结果 DLM组、NLM组双后肢腓肠肌溃疡及缺血明显好转、有明显荧光、肌纤维间毛细血管数多、VEGF表达高,DLC组好转不明显、无荧光、毛细血管数少、VEGF表达少,其差异有统计学意义(P<0.05)。结论脐血内皮祖细胞局部注射治疗糖尿病大鼠下肢缺血有效
Objective To study the effectiveness and mechanism of local injection of endothelial progenitor cells of cord blood for the treatment of diabetic lower limb ischemia and provide a theoretical basis for the clinical treatment of diabetic foot disease. Method 1 The umbilical cord blood mononuclear cells were isolated from mothers’ and their babies and cultured for 7 days. The number of viable cells was counted by flow cytometry. 2 Wistar male rats were divided into three groups: ① diabetic ligation after hind limb femoral artery ligation, local injection of EPC (DLM). ② Diabetic ligation of both hindlimb femoral artery, local injection of PBS (DLC). ③ normal rat ligation after the hindlimb femoral artery ligation, local injection of EPC (NLM). EPCs were labeled with GFP (green fluorescence), the number of capillary vessels was detected by Ⅷ-factor immunohistochemistry, and VEGFmRNA level was detected by RT-PCR. Results The gastrocnemius ulcers and ischemia in both hindlimbs of DLM group and NLM group were markedly improved with obvious fluorescence. The number of capillaries in the muscle fibers was high, the expression of VEGF was high, but there was no obvious change in the DLC group. There was no fluorescence, the number of capillaries was less and VEGF expression was less. The difference was statistically significant (P <0.05). Conclusion Local injection of endothelial progenitor cells from umbilical cord blood can effectively reduce the ischemia of lower extremities in diabetic rats