摘要
目的探讨超声靶向破坏微泡介导肝细胞生长因子(HGF)促进大鼠梗死心肌血管新生的可行性。方法将40只Wistar大鼠制备成心肌梗死模型后随机分为4组:(1)HGF+超声+微泡组(HGF+US/MB);(2)HGF+超声组(HGF+US);(3)HGF+微泡组(HGF+MB);(4)单纯手术组(SA)。于基因转染后14d处死各组大鼠,采用免疫组织化学法(IHC)检测缺血心肌周围CD34表达,并在显微镜下计数心肌内新生微血管密度(MVD)。采用酶联免疫吸附法(ELISA)检测心肌组织内HGF蛋白表达情况,并对心肌内HGF含量与MVD进行相关性分析。结果IHC结果显示,CD34定位于血管内皮细胞胞膜和胞浆,显示为棕黄色或棕褐色颗粒,新生的微血管被染成棕黄色,显微镜下计数每高倍镜视野下的MVD,结果表明在HGF+US/MB组心肌中MVD最高,为(266.9±39.8)个/高倍镜视野,与其他各组比较差异均有显著性(P<0.01)。ELISA结果显示HGF在HGF+US/MB组心肌中表达最高,为(5.54±0.81)ng/g,且前壁表达高于后壁(P<0.05),与其他各组心肌HGF含量相比差异也均有显著性(P<0.01)。相关分析表明,心肌HGF含量与MVD呈明显正相关。结论超声靶向破坏微泡可介导HGF基因在缺血心肌内的高效转移并促进血管新生,为心肌梗死的基因治疗提供了一个新途径。
ABSTRACT： Objective 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 established ： HGF ＋ ultrasound ＋ microbubble （ HGF ＋ US/MB） groups, HGF and ultrasound （ HGF ＋ US） group, HGF and micro- bubble （ HGF ＋ MB） group, and surgery alone （SA） group. Ultrasound-targeted destruction microbubble loaded with HGF gene with ECG trigger was performed in HGF ＋ US/MB group, and HGF gene with ECG trigger was performed in HGF ＋ US group. Microbubble loaded with HGF gene was infused intravenously in HGF ＋ MB group, and normal saline were infused in SA group. All rats were killed 14 days after transfection. The CD34 expression was detected by immunohistochemistry （ IHC）, and microvessel density （MVD） was counted in high power field. The HGF expression on myocardium was detected by ELISA, and the correlation between the contents of HGF and MVD in myocardium was analyzed. Results IHC results showed that CD34 expressions, shown as brown granules, were located on the membrane and endochylema of vascular endothelial cells. The MVD in HGF ＋ US/MB group [ （266.9 ＋ 39.8） /HPF ] were highest among all the groups. The contents of HGF in myocardium were highest in HGF ＋ US/MB group [ （5.54 ＋0.81 ） ng/g], and the contents of HGF in anterior wall were significantly higher than those in posterior wall （ P 〈 O. 05 ） ; the difference was also signif- icant when compared with others groups （ P 〈 O. O1 ）. The correlation analysis showed the contents of HGF was positively correlated with MVD in myocardium. Conclusion Ultrasound-targeted microbubble destruction can effectively deliver HGF into the infracted myocardium and facilitate angiogenesis, which provides a novel way in the gene therapy of myocardial infarction.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2008年第1期5-9,I0001,共6页
Acta Academiae Medicinae Sinicae
基金
国家自然科学基金(30430230)
国家高技术研究发展计划项目(863项目)[国科发财字(2006)501号]
国家教育部高等学校博士学科点专项科研基金(20060631009)~~
关键词
超声微泡
肝细胞生长因子
血管新生
ultrasound microbubble
hepatocyte growth factor
angiogenesis
