Li Ling, Huang Li, Yang Hao, Yin Hong-chao
Department of Pathology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.
Zhonghua Bing Li Xue Za Zhi. 2010 Sep;39(9):621-6.
To explore whether overexpression of P-selectin glycoprotein ligand-1 (PSGL-1) promotes the adhesive ability of endothelial progenitor cells and functionally facilitates neovascularization in mouse model of hindlimb ischemia.
Rat endothelial progenitor cells were transfected with recombinant adenovirus vector encoding human PSGL-1. The mRNA and protein expression levels of PSGL-1 were measured by RT-PCR and Western blot, respectively. The effect of overexpression of PSGL-1 in endothelial progenitor cells was analyzed by adherence assay. Histological examination of skeletal muscle sections retrieved from the mouse ischemic hindlimbs was performed, and the hindlimb blood flow was measured by laser Doppler flow meter.
Adenovirus vector expressing of PSGL-1 gene was successfully constructed with high titer of 3.1 × 10¹¹ pfu/ml. After transfection, PSGL-1 gene was highly expressed in the transfected endothelial progenitor cells. In vitro assay showed that overexpression of PSGL-1 enhanced the adhesive properties of endothelial progenitor cells. When the transfected endothelial progenitor cells were transplanted into the ischemic hindlimb of nude mice, the number of new capillary vessels was (41.0 ± 2.2)/HPF compared to that of (21.0 ± 2.5)/HPF in the negative control group and (10.0 ± 1.6)/HPF in the blank control group (P < 0.01). Furthermore, the blood flow was increased in the experimental group (119.1% ± 7.0%), whereas in the negative control group, it was (93.3% ± 3.0%) and in the blank control group it was (76.3% ± 12.0%), P < 0.01.
Overexpression of PSGL-1 enhances the adhesive and angiogenic properties of endothelial progenitor cells. The approach may provide an effective therapeutic strategy to improve the efficiency of cell-based proangiogenic therapy.
探讨P-选择素糖蛋白配体-1（PSGL-1）过表达是否能促进内皮祖细胞的黏附能力，并在小鼠后肢缺血模型中功能性地促进新生血管形成。
用编码人PSGL-1的重组腺病毒载体转染大鼠内皮祖细胞。分别通过逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测PSGL-1的mRNA和蛋白表达水平。通过黏附试验分析PSGL-1在内皮祖细胞中过表达的作用。对从小鼠缺血后肢获取的骨骼肌切片进行组织学检查，并用激光多普勒血流仪测量后肢血流量。
成功构建了表达PSGL-1基因的腺病毒载体，其滴度高达3.1×10¹¹ pfu/ml。转染后，PSGL-1基因在转染的内皮祖细胞中高表达。体外试验表明，PSGL-1过表达增强了内皮祖细胞的黏附特性。当将转染的内皮祖细胞移植到裸鼠缺血后肢时，新生毛细血管数量在实验组为（41.0±2.2）/高倍视野，阴性对照组为（21.0±2.5）/高倍视野，空白对照组为（10.0±1.6）/高倍视野（P<0.01）。此外，实验组血流量增加（119.1%±7.0%），而阴性对照组为（93.3%±3.0%），空白对照组为（76.3%±12.0%），P<0.01。
PSGL-1过表达增强了内皮祖细胞的黏附及血管生成特性。该方法可能为提高基于细胞的促血管生成治疗效率提供一种有效的治疗策略。
