糖尿病患者动员后外周血单个核细胞促血管新生能力  

Neovascularization Potential of Mobilized Peripheral Mononuclear Cells from Diabetes Patients

作　　者：周斌[1] 顾东生[1] 刘鹏霞[1] 郑翠玲[1] 董春兰[1] 杜伟廷[1] 武开宏[2] 韩忠朝[1] 

机构地区：[1]中国医学科学院中国协和医科大学血液学研究所实验血液学国家重点实验室国家干细胞工程技术研究中心,天津300020 [2]中国医学科学院中国协和医科大学阜外心血管病医院小儿心胸外科,北京100037

出　　处：《中国医学科学院学报》2007年第2期262-267,I0004,共7页Acta Academiae Medicinae Sinicae

基　　金：国家高技术研究发展计划项目(863项目)(2003AA205060);国家重点基础研究发展计划项目(973计划)(001CB5101)~~

摘　　要：目的研究糖尿病患者来源的动员后外周血单个核细胞(M-PBMNCs)移植治疗肢体缺血的疗效,探讨糖尿病患者M-PBMNCs促血管新生能力是否有缺陷及其病理生理机制。方法体外培养内皮祖细胞并鉴定和计数。结扎链脲菌素诱导的糖尿病裸鼠股动脉,造成重度缺血模型后,将来自糖尿病患者、正常人的M-PBMNCs(106)及磷酸盐缓冲液(PBS)分别局部注射到缺血组织中。在第1、3、7、14、21、28天分别用激光多普勒检测局部血流,观察缺血肢体的活动度和缺血情况,用免疫组织化学方法检测缺血部位毛细血管密度和小动脉密度。结果体外内皮祖细胞培养显示糖尿病患者M-PBMNCs中的内皮祖细胞数量相对较少(P<0.05),裸鼠局部缺血组织血流量及血管密度均与糖尿病患者的内皮祖细胞数量呈正相关(R=0.486,P<0.05;R=0.491,P<0.05),而糖尿病患者内皮祖细胞的数量又与其病程呈负相关(R=-0.587,P<0.05)。糖尿病患者M-PBMNCs促进缺血下肢血流和功能恢复的疗效强于PBS组,但不及正常人M-PBMNCs组(P<0.05)。移植了糖尿病患者M-PBMNCs的缺血组织的毛细血管、小动脉密度均低于正常人M-PBMNCs组(P<0.05)。结论糖尿病患者M-PBMNCs的促血管新生能力存在缺陷,使其移植治疗肢体缺血的疗效减低。Objective 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. Methods Endothelial progenitor cells （EPC） were cultured in EGM-2MV, and then characterized by uptake of 1, 1-dioctadecyl-3, 3, 3, 3-tetramethylin-docarbocyanine-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 contrls, 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, and28 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 α-smooth muscle actin （α-SMactin）. Results 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 ischemic 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 ）. Conclusion Diabetes decreased the capability of M-PBMNCs to augment neovascularization in ischemia.

关 键 词：糖尿病 动员后外周血单个核细胞 内皮祖细胞 缺血 

分 类 号：K826.2[历史地理—历史学]