骨髓单个核细胞和骨髓源内皮祖细胞移植促进血流重建的比较研究

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8×10~6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 × 10~6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 4±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.

Comparison of revascularization of bone marrow mononuclear cells and endothelial progenitor cells transplantation

Objective:To compare the effect on revascularization of bone marrow mononuclear cells （BM/MNCs）and endothelial progenitor cells （EPCs） transplantation, and investigate  the roles of non-endothelial progenitor cells in angiogenesis.
Methods:BM/MNCs were harvested from Lewis rats bone marrow. The EPCs were obtained after BM/MNCs were cultured by EBM-2 medium for seven days. Lower limb ischemic model was made on Lewis rats。After limb ischemic model were made, 0.8ml D-Hanks, 8×106 BM/MNCs and 8×106 EPCs were injected into control group (n=6), BM/MNCs group (n=6) and EPCs group (n=6) respectively. At 3 weeks after cells transplantation, the quantity of collateral vascularization was observed with digital subtraction angiography(DSA). Meanwhile, CD31and α-SMA immunohistochemical stain were used to count the capillary and arteriolar density of the excised gastrocnemius muscle of the ischemic limb.
Results:There was no significant difference between BM/MNCs group and EPCs group in the density of the capillaries［（31.67±7.87）/HP vs(32.83±5.38)/HP，P＞0.05］,the number of collaterals ［（4.17±0.75）vs(4.50±1.38)，P＞0.05）］ and density of arterioles ［（4.83±1.47）vs(5.50±2.35)，P＞0.05）］, which were higher than those in control group, respectively.
Conclusions:Non-endothelial progenitor cells in angiogenesis has the same effects as EPC in the treatment of stem cells transplantation for ischemic disease of the limbs.