PEP-1-CAT提高移植入大鼠下肢缺血部位骨髓间充质干细胞的存活率  

PEP-1-CAT Enhances the Viability of MSC Transplanted into Ischemic Lower Limb of Rats

作　　者：张蕾 董小伟 杨建业 郑飞 孔霞 郭凌郧 唐俊明 黄永章 王家宁 

机构地区：[1]湖北医药学院附属人民医院临床医学研究所,湖北十堰442000 [2]湖北医药学院附属人民医院心血管内科,湖北十堰442000

出　　处：《湖北医药学院学报》2012年第5期371-375,封2,共6页Journal of Hubei University of Medicine

基　　金：国家自然科学基金资助项目(81170095);湖北省教育厅优秀中青年创新团队资助计划项目(T200811)

摘　　要：目的:研究转导入骨髓间充质干细胞(Mesenchymal stem cells,MSC)的PEP-1-CAT对MSC存活力的影响及其在大鼠缺血诱导的血管新生中的作用。方法:通过检测Y染色体性别决定区(Sex-determining region of Y-chromosome,SRY)基因的表达和红色细胞膜荧光探针(1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate,DiI)标记的MSC的存留情况,以分析雄性大鼠MSC移植到雌性大鼠下肢缺血部位后的存活状况。MSC在下肢缺血部位的凋亡采用TUNEL法检测。下肢缺血部位血管新生的情况采用血管内皮细胞特异的表面分子CD31的表达进行分析。采用Western blot法检测下肢缺血部位血管内皮生长因子(Vascular endothelial growth factor,VEGF)的表达。结果:PEP-1-CAT预处理的MSC移植到下肢缺血部位后,凋亡细胞减少,存活率比对照组提高了约4倍。PEP-1-CAT预处理组中,下肢缺血区域的CD31表达明显高于对照组,VEGF表达约为对照组的2倍。结论:PEP-1-CAT预处理可增强移植到大鼠下肢缺血部位MSC的存活率,并可通过上调VEGF表达改善损伤部位的血管新生。Objective To investigate the role of PEP-1-CAT transduction on the viability of mesenchymal stem cells(MSC) and its effect on ischemia-induced angiogenesis.Methods The viability of gender-mismatched-transplanted MSC in rat lower limb ischemic muscles was assessed by measuring sex-determining region of Y-chromosome(SRY) expression and evaluating the retained-DiI-labeled cells.MSC apoptosis in ischemic area was determined by TUNEL assay.The effect of PEP-1-CAT-transduced MSC on angiogenesis in vivo was determined by CD31 immunohistochemical staining in the lower limb ischemia model.Vascular endothelial growth factor(VEGF) expression in lower limb ischemia areas was detected by Western blotting.Results The viability of PEP-1-CAT-transduced MSC implanted into ischemic area in rat lower limb ischemia model was increased by four-fold.Moreover,PEP-1-CAT-transduced MSC significantly enhanced ischemia-induced angiogenesis by up-regulating VEGF and CD31 expression,and PEP-1-CAT treatment upregulated VEGF levels about two-fold compared to control.Conclusion PEP-1-CAT-transduction can increase MSC viability and stimulate ischemia-induced angiogenesis by up-regulating VEGF expression.

关 键 词：间充质干细胞 PEP-1-过氧化氢酶 下肢缺血 移植 存活 血管生成 

分 类 号：R543[医药卫生—心血管疾病]