自体骨髓细胞局部移植与粒细胞集落刺激因子皮下注射对缺血区血管生成作用的对比研究

The comparision of the autologous bone marrow mononuclear cell local transplantation and granulocyte colony-stimulating factor for peripheral neovascularization

目的探讨自体骨髓单个核细胞局部移植对后肢缺血区血管新生作用及与经粒细胞集落刺激因子皮下注射对外周血管新生的对比研究。方法20只肢体缺血新西兰白兔分为2组,骨髓细胞局部移植组n(=10)于缺血局部肌肉注射新鲜分离的自体骨髓单个核细胞悬液,局部移植对照组n(=10)局部注射磷酸盐缓冲液;另20只新西兰白兔分为手术组及药物治疗组。药物治疗组n(=10)于结扎并离断左侧股动脉及其分支动脉前24h及术后6d给予G-CSF10μg/k(g.d)皮下注射处理,手术组n(=10)于术前及术后给予等量磷酸盐缓冲液。4组分别于实验前(0d)、7d和28d进行彩色多普勒血流检测,于28d时处死动物,切取内收肌和半膜肌,进行免疫组化法分析,明确毛细血管形态。结果7d和28d时彩色多普勒血流显示,骨髓细胞局部移植组和药物治疗组收缩期血流速度、舒张末期血流速度、血流量、血管阻力指数分别与局部移植对照组及手术组间有显著差异;而上述各指标在骨髓细胞局部移植组和药物治疗组之间无论7d和28d均无统计学意义(P>0.05)。病理检查示骨髓细胞局部移植组和药物治疗组单位面积毛细血管密度分别高于局部移植对照组及手术组(P<0.01),前两者之间无统计学意义。结论G-CSF动员骨髓干细胞能取得自体骨髓细胞局部移植相似的促血管新生作用。

Objective To compare the effect of the autologous bone marrow mononuclear cell（BM-MNCs） local transplantation with granulocyte colony-stimulating factor（G-CSF） on peripheral neovasclurization. Methods In autologous BM-MNCs local transplantation group （n = 10） and control group （n = 10）, autologous BM-MNCs were injected into the ischemic skeletal muscles in rabbit,and phosphate buffered saline administrated for control group. In G-CSF treatment group （n = 10） and control group（n = 10） after unilateral femoral artery and its branches were surgically ligated and excised in rabbits,G-CSF 10 μ4/kg and saline were injected subcutaneously before 24 h of ligation the femoral artery and for 6 days after surgery once daily in G-CSF group and control group. Hemodynamics parameters were measured at day 0,day 7 and day 28 in four groups, respectively. On day 28, the animals were sacrificed in groups for pathological anlysis by NBT/BCIP staining. Results Compared with the control group,color Doppler flow image in BM-LT group was recovered signicantly,and its systolic peak velocity and end -diastolic velocity ,blood volume ,resistance index were also improved. However,no significant difference was found between BM-LT group and G-CSF treatment group in day 7 and day 28 （P 〉 0.05）. Pathological examinations revealed ,a higher capillary density was found in BM-LT group and G-CSF group than that in control group （ P 〈 0.01）. Conclusion The G-CSF treatment can also be beneficial to regenerate vascular structure as the autologous BM-MNCS transplantation,and its effect is superior to the later.