通过移植粒细胞集落刺激因子（G-CSF）刺激的骨髓细胞治疗糖尿病兔的缺血肢体

Treatment of ischemic limbs by transplantation of G-CSF stimulated bone marrow cells in diabetic rabbits.

作者信息

Meng Jianbo, Yao Xiaoguang, Kang Shengqun, Kang Suxian, Yang Yan, Zhou Chenguang

机构信息

Departments of 1Hematology, Hebei Medical University Third Hospital, Ziqiang Road 139, Shijiazhuang 050051, Hebei, China.

出版信息

Clin Invest Med. 2010 Jun 1;33(3):E174-80. doi: 10.25011/cim.v33i3.13723.

DOI:10.25011/cim.v33i3.13723

PMID:20519096

Abstract

PURPOSE

To evaluate the efficacy of autologous transplantation of granulocyte colony-stimulating factor (G-CSF) stimulated bone marrow mononuclear cells in the treatment of diabetic lower limb ischemic disease in a rabbit model.

METHODS

The diabetic model was produced by intravenous injection of 5% alloxan into New Zealand white rabbits. The lower limb ischemia model was created by femoral artery ligation in diabetic rabbits (n=50) 14 days after alloxan injection. Animals were then randomized into five groups (10 rabbits per group): group 1, transplanted with G-CSF immobilized peripheral blood mononuclear cells (PBMNC); group 2, transplanted with G-CSF stimulated bone marrow mononuclear cells (BMMNC); group 3, animals were transplanted with non-stimulated BMMNCs; group 4, G-CSF group injected with G-CSF alone without transplantation; and group 5, PBS group treated with PBS alone. Necrosis of foot or toes and blood flow recovery in ischemic limbs was assessed. Expression of von Willebrand factor (vWF) and vascular endothelial cell growth factor (VEGF) were measured in ischemic muscles by immunohistochemistry analysis.

RESULTS

Blood flow in ischemic limbs was significantly improved in the G-BMMNC group (ratio of blood flows: 0.82+/-0.06) in comparison with the G-PBMNC (0.61+/-0.09) and BMMNC (0.62+/-0.08) groups (P < 0.001). In the G-BMMNC group, the capillary density, a measure of vWF expression, was significantly higher than in either the G-PBMNC or BMMNC groups (47.9+/-2.51 vs. 36.8+/-4.16 and 39.6+/-2.72, respectively, P < 0.05). The expression of VEGF in G-BMMNC animals was significantly increased in comparison with the G-PBMNC and BMMNC groups (16.93+/-0.70 vs. 11.83+/-0.98 and 12.32+/-0.96, respectively, P < 0.05).

CONCLUSION

A combination of G-CSF stimulation and autologous transplantation of bone marrow stem cells synergistically improved neovascularization and angiogenesis in ischemic limb tissues in diabetic rabbits.

摘要

目的

评估粒细胞集落刺激因子（G-CSF）刺激的骨髓单个核细胞自体移植治疗兔糖尿病下肢缺血性疾病的疗效。

方法

通过向新西兰白兔静脉注射5%四氧嘧啶建立糖尿病模型。在注射四氧嘧啶14天后，通过结扎糖尿病兔（n = 50）的股动脉建立下肢缺血模型。然后将动物随机分为五组（每组10只兔）：第1组，移植G-CSF固定的外周血单个核细胞（PBMNC）；第2组，移植G-CSF刺激的骨髓单个核细胞（BMMNC）；第3组，移植未刺激的BMMNC；第4组，G-CSF组仅注射G-CSF而不进行移植；第5组，PBS组仅用PBS治疗。评估足部或脚趾的坏死情况以及缺血肢体的血流恢复情况。通过免疫组织化学分析测量缺血肌肉中血管性血友病因子（vWF）和血管内皮细胞生长因子（VEGF）的表达。

结果

与G-PBMNC组（0.61±0.09）和BMMNC组（0.62±0.08）相比，G-BMMNC组缺血肢体的血流显著改善（血流比值：0.82±0.06）（P < 0.001）。在G-BMMNC组中，作为vWF表达指标的毛细血管密度显著高于G-PBMNC组或BMMNC组（分别为47.9±2.51对36.8±4.16和39.6±2.72，P < 0.05）。与G-PBMNC组和BMMNC组相比，G-BMMNC组动物中VEGF的表达显著增加（分别为16.93±0.70对11.83±0.98和12.32±0.96，P < 0.05）。