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
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张建E-mail:tongzhu009@163.com

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国家高技术研究发展计划(863) 基金资助项目(2006AA02A134);北京市自然科学基金资助项目(7072031);北京市委组织部优秀人才基金资助项目(20061D0501800265)。


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

    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。

    Abstract:

    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.

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佟铸, 谷涌泉, 李建新, 罗涛, 张淑文, 张建.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[J].中国普通外科杂志,2009,18(12):1259-1262.
DOI:10.7659/j. issn.1005-6947.2009.12.008

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  • 收稿日期:2009-06-19
  • 最后修改日期:2009-09-20
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  • 在线发布日期: 2009-12-15