Zhou Bin, Gu Dong-Sheng, Liu Peng-Xia, Zheng Cui-Ling, Dong Chun-Lan, Du Wei-Ting, Wu Kai-Hong, Han Zhong-Chao
National Key Laboratory of Experimental Hematology, National Research Center for Stem Cell Engineering and Technology, Institute of Hematology, CAMS and PUMC, Tianjin 300020, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2007 Apr;29(2):262-7.
To determine whether mobilized peripheral blood mononuclear cells (M-PBMNCs) obtained from patients with diabetes was impaired in therapeutic neovascularization in limb ischemia, and to explore the pathological mechanisms of the impairment.
Endothelial progenitor cells (EPC) were cultured in EGM-2MV, and then characterized by uptake of 1, 1-dioctadecyl-3, 3, 3, 3-tetramethylindocarbocyanine-labeled acetylated low density lipoprotein (Dil-AcLDL) and binding of ulex europaeus agglutinin (UEA). The number of EPC was compared between M-PBMNCs obtained from diabetic patients and those from normal subjects. M-PBMNCs obtained from diabetic patients, M-PBMNCs obtained from normal controls, or PBS were injected into the ischemic limbs of streptozotocin-induced diabetic nude mice. The limb blood perfusion was detected by laser Doppler blood perfusion imaging between these three groups in the following 1, 3, 7, 14, 21, and 28 days. Ambulatory score and ischemia damage were evaluated in the following 4 weeks. Capillary/fiber ratio was detected by CD31 or BS-1 lectin, and arteriole density was detected by alpha-smooth muscle actin (alpha-SMactin).
The number of EPC from diabetic patients were positively correlated with the blood perfusion (R = 0.486, P < 0.05) and capillary density (R = 0.491, P < 0.05), and the EPC number in diabetic patient were negatively correlation with their disease courses (R = - 0.587, P < 0.05). Transplantation of diabetic M-PBMNCs augmented the blood perfusion of ischemia hindlimbs, increased the capillary and arteriole densities, and promoted the collateral vessel formation. However, all the improvements were less significant in the diabetic patients than in the non-diabetic patients (P < 0.05).
Diabetes decreased the capability of M-PBMNCs to augment neovascularization in ischemia.
确定从糖尿病患者获取的动员外周血单个核细胞（M-PBMNCs）在肢体缺血治疗性血管新生中是否受损，并探讨这种损伤的病理机制。
在内皮细胞生长培养基-2MV（EGM-2MV）中培养内皮祖细胞（EPC），然后通过摄取1,1-二油酰基-3,3,3,3-四甲基吲哚羰花青标记的乙酰化低密度脂蛋白（Dil-AcLDL）和荆豆凝集素（UEA）结合进行鉴定。比较糖尿病患者和正常受试者获取的M-PBMNCs中EPC的数量。将糖尿病患者获取的M-PBMNCs、正常对照获取的M-PBMNCs或磷酸盐缓冲液（PBS）注射到链脲佐菌素诱导的糖尿病裸鼠的缺血肢体中。在接下来的1、3、7、14、21和28天通过激光多普勒血流灌注成像检测这三组的肢体血流灌注。在接下来的4周评估动态评分和缺血损伤。通过CD31或BS-1凝集素检测毛细血管/纤维比率，通过α-平滑肌肌动蛋白（α-SMactin）检测小动脉密度。
糖尿病患者的EPC数量与血流灌注呈正相关（R = 0.486，P < 0.05）和毛细血管密度呈正相关（R = 0.491，P < 0.05），糖尿病患者的EPC数量与病程呈负相关（R = -0.587，P < 0.05）。移植糖尿病M-PBMNCs可增加缺血后肢的血流灌注，增加毛细血管和小动脉密度，并促进侧支血管形成。然而，糖尿病患者的所有改善均不如非糖尿病患者显著（P < 0.05）。
糖尿病降低了M-PBMNCs在缺血中增强血管新生的能力。
