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目的探讨AngⅡ2型受体(AT2R)基因局部电穿孔转染对大鼠颈总动脉球囊损伤后新生内膜增生的影响。方法建立大鼠颈总动脉球囊损伤模型,用局部电穿孔方法转染AT2R真核表达载体(pEGFP-AT2R)或空载体(pEGFP-N2),分别于术后7、14 d和21 d采用HE染色及RT-PCR方法进行AT2R、AngⅡ1型受体(AT1R)在颈动脉壁中表达的变化和组织形态学分析,检测其对在体血管新生内膜的影响作用。结果球囊损伤21 d后,pEGFP-AT2R组大鼠颈动脉AT2R mRNA表达为(1.262±0.317),pEGFP-N2组为(0.396±0.100),单纯损伤组为(0.410±0.053),pEGFP-AT2R组与pEGFP-N2组和单纯损伤组相比差异有统计学意义(P<0.01);球囊损伤后21 d时AT1R的表达,pEGFP-AT2R组为(0.469±0.065)、pEGFP-N2组为(0.363±0.046)、单纯损伤组为(0.373±0.045),pEGFP-AT2R组较pEGFP-N2组和单纯损伤组相比差异有统计学意义(P<0.05),pEGFP-N2组和单纯损伤组间无统计学差异(P>0.05);球囊损伤后21 d,pEGFP-AT2R组的内膜面积与中膜面积比(I/M)为(0.828±0.101),pEGFP-N2组为(1.432±0.086),单纯损伤组为(1.515±0.078),pEGFP-AT2R组较pEGFP-N2组和单纯损伤组相比差异有统计学意义(P<0.01)。pEGFP-AT2R组与单纯损伤组相比,使实验动物新生内膜增生平均受抑制率达45.35%。结论在体血管局部电穿孔转染AT2R基因可使AT2R在损伤血管组织表达较单纯损伤组明显增加。AT2R基因与AT1R基因之间在表达量上不存在此消彼长的关系。
Objective To investigate the effect of local electroporation of angiotensin Ⅱ type 2 receptor (AT2R) gene on neointimal hyperplasia after carotid artery balloon injury in rats. Methods A rat model of common carotid artery balloon injury was established. AT2R eukaryotic expression vector (pEGFP-AT2R) or empty vector (pEGFP-N2) was transfected by electroporation. The rabbits were sacrificed at 7, 14, The changes of AT2R and AngⅡ1 receptor (AT1R) expression in the carotid artery wall and histomorphological analysis were detected by HE staining and RT-PCR. Results After balloon injury for 21 days, the expression of AT2R mRNA in carotid arteries of pEGFP-AT2R group was (1.262 ± 0.317), that of pEGFP-N2 group was (0.396 ± 0.100), that of pEGFP-AT2R group was (0.410 ± 0.053) (P <0.01). The expression of AT1R at 21 d after balloon injury was (0.469 ± 0.065) in pEGFP-AT2R group and (0.363 ± 0.046) and simple injury group (0.373 ± 0.045) respectively. There was significant difference between pEGFP-AT2R group and pEGFP-N2 group and simple injury group (P <0.05) There was no significant difference between the two groups (P> 0.05). On day 21 after balloon injury, the ratio of intima to media was (0.828 ± 0.101) in pEGFP-AT2R group and (1.432 ± 0.086), and the injury group was (1.515 ± 0.078). There was significant difference between pEGFP-AT2R group and pEGFP-N2 group and injury group (P <0.01). The inhibition rate of neointimal hyperplasia in experimental animals was 45.35% on average in pEGFP-AT2R group compared with simple injury group. Conclusion Transfection of AT2R gene by local electroporation in vivo can significantly increase the expression of AT2R in injured vascular tissue compared with simple injury group. There is no reciprocal relationship between the AT2R gene and the AT1R gene.