论文部分内容阅读
目的研究重组腺病毒介导肝细胞生长因子(adenoviral vector mediated human hepatocyte growth factor,AdHGF)修饰的胎盘源间充质干细胞(placenta-derived mesenchymal stem cells,PMSC)在兔肢体缺血模型中促新生血管生成的作用。方法无菌取足月产妇胎盘胎儿面中心区域胎盘组织,胶原酶消化法分离干细胞,体外培养传代,感染Ad-HGF 48 h后收集细胞用于治疗兔肢体缺血实验。左侧治疗组后肢缺血肌肉组织内多点注射总细胞数5×106/ml Ad-HGF修饰的PMSC,右侧对照组后肢缺血肌肉组织内注射生理盐水。结果治疗后第14天腹主动脉穿刺行数字减影血管造影(digital subtraction angiography,DSA)检查,可见左侧肢体缺血治疗后微血管生成数明显多于未治疗右侧肢体,血管形成能力明显增强。苏木精伊红染色法(HE)后置光镜下观察显示,治疗组毛细血管密度明显大于未治疗组(P<0.05)。实时荧光定量聚合酶链反应(quantitative polymerase chain reaction,q PCR)检测后肢肌肉组织中血管内皮生长因子(vascular endothelial growth factor,VEGF)、碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)、HGF的表达明显升高(P<0.05)。结论经Ad-HGF基因修饰的PMSC能明显促进血管新生,加快局部缺血组织血流循环的重建,是治疗肢体慢性缺血的有效手段。
Objective To investigate the effect of recombinant adenovirus vector-mediated human hepatocyte growth factor (AdHGF) -mediated placenta-derived mesenchymal stem cells (PMSC) on promoting angiogenesis in rabbit limb ischemia model Generated role. Methods Placenta of placenta in placenta fetus of aphasia was collected by aseptic technique. Stem cells were isolated by collagenase digestion and subcultured. Ad-HGF cells were collected for 48 h and then used to treat rabbits limb ischemia. In the left hindlimb ischemia group, PMSC with 5 × 106 / ml Ad-HGF modified cells was injected into the ischemic hindlimb muscle of the left hind limbs, and saline was injected into the muscle tissue of the right hindlimb ischemia muscle of the right control group. Results The digital subtraction angiography (DSA) was performed on abdominal aorta puncture on the 14th day after treatment. The number of angiogenesis in the left limb after ischemia treatment was significantly higher than that in the untreated right limb and angiogenesis was significantly enhanced . Hematoxylin and eosin staining (HE) post-light microscopy showed that the treatment group, capillary density was significantly greater than the untreated group (P <0.05). Real-time quantitative polymerase chain reaction (q PCR) was used to detect the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b FGF) , HGF expression was significantly increased (P <0.05). Conclusions PMSC modified by Ad-HGF gene can significantly promote angiogenesis and accelerate the reperfusion of blood flow in ischemic tissue. It is an effective method to treat chronic ischemia of limbs.