Experimental research on therapeutic angiogenesis induced by hepatocyte growth factor directed by ultrasound-targeted microbubble destruction in rats
Experimental research on therapeutic angiogenesis induced by hepatocyte growth factor directed by ultrasound-targeted microbubble destruction in rats
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DOI：
10.7863/jum.2008.27.3.453
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发表时间：
2008-03-01
影响因子：
2.3
通讯作者：
Xu, Chuanshan
中科院分区：
文献类型：
--
作者：
Li, Xingsheng;Wang, Zhigang;Xu, Chuanshan
Objective.本研究旨在探讨超声靶向破坏微泡介导肝细胞生长因子（HGF）诱导心肌梗死血管新生的可行性。将40只Wistar大鼠随机分为4组：（1）HGF+超声+微泡（HGF+US/MB）组，（2）HGF+超声组，（3）HGF+微泡组，（4）单纯手术组。在HGF+US/MB组中，使用心电图触发模式破坏载有HGF基因的超声靶向微泡。转染14天后处死大鼠。荧光显微镜下观察各组大鼠心肌、肝脏、肾脏绿色荧光蛋白表达增强;免疫组化法检测CD 34表达，高倍镜下计数微血管密度（MVD）;免疫印迹法和酶联免疫吸附法检测心肌组织中肝细胞生长因子的表达。HGF+US/MB组心肌绿色荧光蛋白表达增强，但仅在小血管和毛细血管内皮中检测到少量区域的HGF表达，而单纯手术组和HGF+微泡组均未检测到HGF表达。显微镜下MVD计数结果显示，HGF+US/MB组心肌中MVD最高。免疫印迹和酶联免疫吸附试验结果显示，HGF+US/MB组心肌中HGF含量最高。超声靶向破坏微泡可将HGF导入梗死心肌并产生血管生成效应，为心肌梗死的基因治疗提供了新的思路。
Objective. The purpose of this study was to explore the feasibility of therapeutic angiogenesis in myocardial infarction induced by hepatocyte growth factor (HGF) mediated by ultrasound-targeted microbubble destruction.Methods. Forty Wistar rats were divided into 4 groups after the models of myocardial infarction were prepared: (1) HGF, ultrasound, and microbubbles (HGF+US/MB), (2) HGF and ultrasound, (3) HGF and microbubbles, and (4) surgery alone. Destruction of ultrasound-targeted microbubbles loaded with the HGF gene with an electrocardiographic trigger mode was performed in the HGF+US/MB group. All the rats were killed after being transfected for 14 days. Enhanced green fluorescent protein expression was examined in the myocardium, liver, and kidney in all groups by fluorescence microscopy; CD34 expression was detected by immunohistochemistry, and microvessel density (MVD) was counted in the high-power field on microscopy, Hepatocyte growth factor expression in the myocardium was detected by western blotting and an enzyme-linked immunosorbent assay.Results. Enhanced green fluorescent protein expression was detected in the myocardium of the HGF+US/MB group, but a few areas of HGF expression were detected only in small vessels and the capillary endothelium, and no expression was found in the surgery-alone and HGF and microbubbles groups. The results of MVD counting by microscopy showed that the MVD in the myocardium of the HGF+US/MB group was the highest among all the groups. The results of western blotting and the enzyme-linked immunosorbent assay showed that the amount of HGF in the myocardium was highest in the HGF+US/MB group.Conclusions. Ultrasound-targeted microbubble destruction could deliver HGF into the infracted myocardium and produce an angiogenesis effect, which could provide a novel strategy for gene therapy of myocardial infarction.