目的观察腺病毒介导的血红素加氧酶-1(HO—1)基因转染大鼠后对脑缺血再灌注损伤的保护作用。方法雄性SD大鼠随机分为4组:假手术对照组(SH)、生理盐水组(V)、空载体组(Ad)和Ad—HO-1转染组(HO)。后三组在缺血前3d于右侧脑室部分别注射20μl生理盐水、含1μl空载体腺病毒(1.0×10^10plaque-forming unit/ml,PFU/ml)的生理盐水或含1μl重组HO-1腺病毒(1.0×10^10PFU/ml)的生理盐水,连续注射3d后,采用右侧大脑中动脉栓塞法(MCAO)建立脑缺血再灌注模型。每组大鼠测定神经功能后,处死大鼠并取全脑标本,测定右脑梗死体积及细胞凋亡指标,荧光显微镜下观察脑组织荧光蛋白的表达情况,Western blot检测脑组织HO-1的表达。结果HO组中HO-1表达量明显高于Ad组和V组,Ad组和HO组可见有荧光蛋白表达,转染率为34.5%±3.4%。HO组神经功能显著优于Ad组和V组(P〈0.001)。与SH组比较,V组、Ad组脑梗死体积、神经细胞凋亡明显升高。与V组及Ad组比较,HO组脑梗死体积显著减小(P〈0.01)、神经元凋亡显著减少(P〈0.01)。结论腺病毒携带的HO-1基因能有效的转染脑组织,并在脑内稳定表达;HO-1基因转染显著减轻脑缺血再灌注后神经细胞损伤。
Objective To observe the protective effect on cerebral ischemia-repeffusion injury after transfection of heme oxygenase-1 (HO-1) gene in rat. Methods SD rats were randomly divided into four groups: sbam group (SH), vehicle group ( V ), empty adenovirus vector group (Ad) and Ad-HO- 1 transfection group (HO). Rats of V, Ad and Ad-HO-1 groups were respectively injected with 20 μl saline, 20 μl saline containing 1 μl of empty vector adenovirus or 1 μl of recombinant HO-1 adenovirus, through the right ventricle for 3 days before ischemia. Then, cerebral ischemia-reperfusion was estabhshed by middle cerebral artery occlusion (MCAO). After determination of neurological function, the rats were sacrificed and brain specimens were taken to measure the infarction volume and apoptosis, and the expression of fluorescent protein in the brain tissues was observed. The expression of HO-1 in brain tissues was detected by Western blot. Results Westem blot analysis showed that the expression of HO-1 in HO group was significantly higher than that of Ad group and V group. Huorescent protein expression was seen in brain tissues in beth Ad group and HO group, and the transfection rate was 34.5% ±3.4%. The neurological function scores of HO group were significantly higher than that of Ad group and V group (P 〈0.001). Compared with SH group, the volume of cerebral infarction and the neuronal apoptosis in V group and Ad group increased (P 〈0.01 ). Compared with V group or Ad group, the volume of cerebral infarction and the apoptesis in HO group significantly reduced (P 〈0.01 ). Conclusion The HO-1 gene carried by adenovirus can be effectively transfected into the brain tissues and expressed stably in vivo. Transfection of HO-1 gene can significantly reduce the death of neuronal cells after cerebra ischemia-reperfusion.