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目的探讨CT导引下肝细胞生长因子(HGF)基因治疗脑缺血的可行性。方法采用基因重组技术构建携带HGF基因的真核表达质粒,通过脂质体介导法,在CT灌注扫描图像定位下将其多点注射到大鼠急性脑梗死模型的缺血半暗带区域;转染7 d后断颈取脑,切片观察HGF基因于大鼠脑内的表达情况及其生物学效应。结果酶切鉴定及基因测序证实,HGF基因片段已克隆到PIRES2-EGFP的BamH I和Sal I位点之间。HGF基因转染大鼠缺血半暗带区7 d后免疫组化方法证实实验组大鼠转染局部已有HGF表达,血管数量明显多于对照组(P<0.01);CT灌注显示其梗死侧半球的脑血流量高于对照组(P<0.01);TTC染色显示实验组大鼠脑梗死体积小于对照组(P<0.01)。结论脂质体转染HGF基因能够在缺血半暗带区表达,其表达产物能够发挥生物学效应并促进局部侧支循环形成,从而改善脑缺血。
Objective To investigate the feasibility of CT-guided hepatocyte growth factor (HGF) gene therapy for cerebral ischemia. Methods The eukaryotic expression plasmid carrying HGF gene was constructed by gene recombination technique and then injected into the ischemic penumbra of acute cerebral infarction rat model by liposome - mediated method under CT perfusion scan image localization. Seven days after transfection, the brain was cut off from the neck, and the expression of HGF gene in the brain of rats was observed. The biological effects were also observed. Results The restriction enzyme digestion and gene sequencing confirmed that the HGF gene fragment was cloned into the BamH I and Sal I sites of PIRES2-EGFP. HGF gene was transfected into the ischemic penumbra of rats for 7 days, and the expression of HGF in the experimental group was confirmed by immunohistochemistry. The numbers of blood vessels in the experimental group were significantly more than those in the control group (P <0.01) Cerebral blood flow in the hemisphere was higher than that in the control group (P <0.01). TTC staining showed that the volume of cerebral infarction in the experimental group was smaller than that in the control group (P <0.01). Conclusion The liposome-transfected HGF gene can be expressed in the ischemic penumbra and its expression products can exert biological effects and promote the formation of local collateral circulation, thus improving cerebral ischemia.