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目的比较骨髓单个核细胞(MNCs)和骨髓源内皮祖细胞(EPCs)移植促进血流重建的效果,探讨非内皮祖细胞在血流重建中的作用。方法获取Lewis大鼠骨髓MNCs,部分MNCs在体外诱导分化为EPCs。采用Lewis大鼠建立单侧后肢缺血模型。建模后3 d,将模型鼠随机分为3组:(1)对照组(n=6),将0.8 mL D-Hank′s液注入对照组大鼠缺血侧后肢;(2)MNC组(n=6),将8×106个骨髓MNC植入MNC组大鼠缺血侧后肢;(3)EPC组(n=6),将体外培养的8×106个EPC植入EPC组大鼠缺血侧后肢。细胞移植后3周行大鼠后肢动脉造影,检测缺血侧后肢侧支血管数;切取缺血侧后肢腓肠肌,分别行CD31和α-SMA免疫组化染色,计算毛细血管密度和小动脉密度。结果MNC组毛细血管密度与EPC组差异无统计学意义[(31.67±7.87)个/HP vs.(32.83±5.38)个/HP,P>0.05],而均高于对照组(19.67±4.80个/HP)(P<0.05);MNC组侧支血管数与EPC组差异无统计学意义[(4.17±0.75)个vs.(4.50±1.38)个,P>0.05],但均高于对照组[(2.50±1.5)个](P<0.05);MNC组小动脉密度与EPC组差异无统计学意义[(4.83±1.47)个vs.(5.50±2.35)个,P>0.05],亦均高于对照组[(2.17±0.98)个](P<0.05)。结论在骨髓干细胞移植治疗肢体缺血性疾病中,非内皮祖细胞在血流重建中所起的作用与EPC相似。
Objective To compare the effects of bone marrow mononuclear cells (MNCs) and bone marrow-derived endothelial progenitor cells (EPCs) on the reconstruction of blood flow and to explore the role of non-endothelial progenitor cells in blood flow reconstruction. Methods Bone marrow MNCs from Lewis rats were obtained. Some MNCs were induced to differentiate into EPCs in vitro. Lewis rats were used to establish a unilateral hindlimb ischemia model. Three days after modeling, the model rats were randomly divided into three groups: (1) control group (n = 6), 0.8 mL D-Hank’s solution was injected into the ischemic hindlimb of control rats; (2) MNC group (n = 6). 8 × 106 bone marrow MNCs were implanted into ischemic hindlimbs of MNC rats. (3) EPCs (n = 6) Ischemic hindlimb. The hindlimb arterial angiography was performed 3 weeks after cell transplantation to detect the number of collateral vessels in the ischemic hindlimb. The gastrocnemius muscle of the ischemic hindlimb was cut out and the CD31 and α-SMA immunohistochemical staining were performed to calculate the capillary density and arteriolar density. Results There was no significant difference in capillary density between MNC group and EPC group [(31.67 ± 7.87) / HP vs. (32.83 ± 5.38) /HP, P> 0.05], but higher in MNC group than in control group (19.67 ± 4.80 /HP)(P<0.05). The number of collateral vessels in MNC group was not significantly different from that in EPC group [(4.17 ± 0.75) vs (4.50 ± 1.38), P> 0.05], but higher than those in control group [(2.50 ± 1.5)] (P <0.05). There was no significant difference in arteriolar density between MNC group and EPC group (4.83 ± 1.47 vs. 5.50 ± 2.35, P> 0.05) Higher than the control group [(2.17 ± 0.98)] (P <0.05). Conclusions In bone marrow stem cell transplantation for the treatment of limb ischemic disease, the function of non-endothelial progenitor cells in blood flow reconstruction is similar to that of EPC.