Song Ye, Xie Xiaoyun, Gao Yuan, Gu Guojun, Wang Peijun
Department of Ultrasound, Shanghai Tongji Hospital of Tongji University Shanghai 200065, China.
Department of Geriatrics, Shanghai Tongji Hospital of Tongji University Shanghai 200065, China.
Am J Transl Res. 2016 Jun 15;8(6):2585-96. eCollection 2016.
This study aimed to investigate whether ultrasound-induced microbubble destruction was able to promote targeted delivery of adipose-derived stem cells (ASCs) to improve hind-limb ischemia of diabetic mice. Ischemia was induced in the lower limb of db/db mice which were then randomly divided into 5 groups: PBS group, Sham group, ultrasound + microbubble group (US+MB), US+MB+ASCs group and ASCs group. Contrast-enhanced ultrasound perfusion imaging showed the ratio of blood flow in ischemic hind-limb to that in contralateral limb increased over time in five groups. A significant enhancement in US+MB+ASCs group was observed compared with US+MB group (P<0.01). Immunofluorescence microscopy of hind-limb muscle showed the microvessel density (microvessels/skeletal muscle fibers) and arteriolar density in US+MB+ASCs group were higher than in US+MB group, and significantly higher than in other control groups (P<0.01). Masson staining indicated the degree of muscle fibrosis in US+MB+ASCs group was lower than in US+MB. 3 and 7 days after therapy, ELISA and RT-PCR showed the expression of VEGF, P-selectin, ICAM-1 and SDF-1 in US+MB+ASCs group was higher than in US+MB group, and dramatically increased as compared to other groups (P<0.01). 3 and 7 days after therapy, Western blot assay showed the protein expression of P-P13K, P-AKT, VEGF, P-selectin, ICAM-1 and SDF-1 in US+MB+ASCs group was higher than US+MB group (P<0.01). The bioeffects of ultrasound-induced microbubble cavitation is able to up-regulate the expression of pro-inflammatory cytokines, which may improve the targeted delivery, adhesion and paracrine of ASCs, attenuating the hind-limb ischemia in diabetic mice.
本研究旨在探讨超声诱导的微泡破坏是否能够促进脂肪来源干细胞（ASC）的靶向递送，以改善糖尿病小鼠的后肢缺血。对db/db小鼠的下肢进行缺血诱导，然后将其随机分为5组：PBS组、假手术组、超声+微泡组（US+MB）、US+MB+ASC组和ASC组。超声造影灌注成像显示，五组缺血后肢与对侧肢体的血流比值均随时间增加。与US+MB组相比，US+MB+ASC组有显著增强（P<0.01）。后肢肌肉的免疫荧光显微镜检查显示，US+MB+ASC组的微血管密度（微血管/骨骼肌纤维）和小动脉密度高于US+MB组，且显著高于其他对照组（P<0.01）。Masson染色表明，US+MB+ASC组的肌肉纤维化程度低于US+MB组。治疗后3天和7天，ELISA和RT-PCR显示，US+MB+ASC组中VEGF、P-选择素、ICAM-1和SDF-1的表达高于US+MB组，且与其他组相比显著增加（P<0.01）。治疗后3天和7天，蛋白质印迹分析显示，US+MB+ASC组中P-P13K、P-AKT、VEGF、P-选择素、ICAM-1和SDF-1的蛋白质表达高于US+MB组（P<0.01）。超声诱导的微泡空化的生物效应能够上调促炎细胞因子的表达，这可能改善ASC的靶向递送、黏附和旁分泌，减轻糖尿病小鼠的后肢缺血。
