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目的研究脐血内皮祖细胞(EPC)治疗糖尿病下肢缺血的有效性及机制,为临床治疗糖尿病足病(DF)提供理论依据。方法取产妇足月产脐血分离单核细胞,培养7d,流式细胞仪鉴定细胞,并计数活细胞数。将Wistar雄性大鼠分成3组:(1)糖尿病大鼠射线照射后结扎双后肢股动脉,尾静脉注射血管EPC(DE组)。(2)糖尿病大鼠结扎双后肢股动脉,尾静脉注射缓冲液(DP组)。(3)正常大鼠射线照射后结扎双后肢,尾静脉注射EPC(NC组)。用绿色荧光(GFP)示踪EPC,Ⅷ因子免疫组化检测肌纤维间毛细血管数,RT-PCR检测双后肢肌肉血管内皮生长因子(VEGF)mRNA水平。结果 DE、NC组右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,DP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EPC治疗糖尿病大鼠下肢缺血有效。
Objective To study the efficacy and mechanism of cord blood endothelial progenitor cells (EPC) in the treatment of diabetic lower limb ischemia and provide a theoretical basis for the clinical treatment of diabetic foot disease (DF). Methods Umbilical cord blood mononuclear cells were isolated from maternal and fetal umbilical cord blood and cultured for 7 days. The number of viable cells was counted by flow cytometry. The Wistar male rats were divided into three groups: (1) The diabetic rats were irradiated with double hind limb femoral artery and the tail vein was injected with EPCs (DE group). (2) Diabetic rats were ligated with both hindlimb femoral and tail vein injection buffer (DP group). (3) After normal irradiation, both hindlimbs were ligated and EPCs were injected into caudal vein (NC group). EPCs were labeled with green fluorescence (GFP), the number of capillary vessels was detected by Ⅷ factor immunohistochemistry, and the level of vascular endothelial growth factor (VEGF) mRNA was detected by RT-PCR. Results The gastrocnemius ulcers and ischemia in the right hind limbs of DE and NC groups were significantly improved, with significant fluorescence. The number of capillaries in the muscle fibers was high, the expression of VEGF was high, the changes in the DP group were insignificant, no fluorescence, few capillaries, and less VEGF expression There was statistical significance (P <0.05). Conclusion Umbilical cord blood EPC is effective in treating lower limb ischemia in diabetic rats.