超声评价兔骨髓间充质干细胞自体移植对缺血肢体血管再生的影响

摘要

Objective To investigate the significance and effect of ultrasonic diagnosis on the autologous bone marrow mesenchymal stem cells (MSC) in angiogenesis. Methods Twenty-four New Zealand rabbits were divided into experiment group (12) and the control group (12). Then rabbit bone marrow MSCs from experiment group were isolated, caltured and marked with Brdu. After ischemic hind limb animal model on all rabbits was set up, autologous bone marrow MSCs were directly injected into the ischemic hind limb muscles in experiment group while same volume normal saline was used in the control group. Two weeks after the implantation of autologous bone marrow MSCs, 2D and color Doppler flow imaging (CDFI) detection were used in rabbit femoral artery of the two groups to observe the inner diameter of the blood vessel, the peak velocity and the acceleration time. The disposition of transplaned cells and the state of angiogenesis in ischemic muscles were assessed using immunofluorescence staining. Results The results of 2D and Doppler ultrasound detection showed the inner diameter of the blood vessel and the peak velocity of the blood current in experiment group obviously higher than that of the control group , and the acceleration time was obviously smaller than that of the control group P<0.01. The immunofluorescence staining showed there were transplanted cells existed in transplanted portion and state of angiogenesis was supurior obviously than that of the control. Conclusions Bone marrow MSCs had the effect to promote angiogenesis. Implantation of autologous bone marrow MSCs was a simple and efficient therapeutic method for the ischemia hind limb. Using high-frequency ultrasound to detect femoral artery may provide a practical and useful method to evaluate the effect on implanted autologous bone marrow mesenchymal stem cells.%目的 探讨超声在评价兔骨髓间充质干细胞(BMSCs)自体移植促进血管再生过程中的应用价值.方法 把24只新西兰兔分为实验组(12只)和对照组(12只),分离培养实验组兔骨髓间充质干细胞,Brdu标记,建立兔后肢缺血动物模型,将取于自体的骨髓间充质干细胞制成细胞悬液注射于实验组兔缺血部位,对照组注射等量生理盐水.2周后对实验组和对照组兔股动脉及股浅动脉起始段行常规超声及彩色多普勒血流成像(CDFI)检测,检测股动脉血管内径、血流峰值速度及血流加速时间;取缺血部位肌肉组织,行免疫组织化学染色检测实验组移植细胞存在分布情况以及病理学检查血管再生情况.结果 骨髓间充质干细胞移植实验2周后,实验组兔股动脉内径、血流峰值速度均大于对照组,血流加速时间小于对照组 (P＜0.01).免疫组织化学染色结果显示移植部位有移植细胞存在,实验组血管再生情况明显优于对照组.结论 骨髓间充质干细胞移植具有促进血管生成的作用.自体移植有望成为一种简单且有效治疗下肢缺血的方法.高频超声检测股动脉对自体移植效果评价提供了有效实用的方法.