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目的观察阳离子脂质体混合物与β1受体反义寡核苷酸(β1-AS-ODN)单次注射对两肾一夹肾性高血压大鼠血压及心脏β受体mRNA、受体密度的影响。方法用SD大鼠制作两肾一夹肾性高血压模型,将动物随机分为5组,每组18只:β1-AS-ODN组,β1-IN-ODN组,2KIC组,假手术组,SD组。于术后第4周经鼠尾静脉注射阳离子脂质体/β1-ODN的混合物(摩尔比2.0),注射剂量为0.5mg·kg~(-1),Tail-cuff法检测血压,半定量RT-PCR测定β1mRNA水平,放射免疫法测定受体密度,Western blot测定蛋白水平。数据采用one-way ANOVA分析,两两比较用Newman-Keuls法。结果与2KIC组相比,β1-AS-ODN可使血压下降维持4周(P<0.05),可以使血压最大下降39 mmHg;β1-AS-ODN组与高血压组β1受体mRNA水平无明显差异,但β1受体蛋白水平明显降低(P<0.05);与2K1C组比较,注射后第2、7、30天反义组的β1受体水平明显降低,分别为29.5%、35.5%、20.4%(P<0.01,P<0.01,P<0.05);三组β2受体水平差异无统计学意义。结论以β1受体为靶基因的反义基因治疗降压效果显著,β1反义寡核苷酸在转录后发挥作用抑制目的基因表达;β1反义寡核苷酸的降压效果与其特异性地和心脏β1受体结合相关,而与β2受体无关。
Objective To observe the effect of single injection of β1-AS-ODN mixture and cationic liposome mixture on the blood pressure and β receptor mRNA and receptor density in two-kidney-one-folder hypertensive rats influences. Methods Two kidneys and one kidneys hypertensive model were established in SD rats. The animals were randomly divided into 5 groups with 18 rats in each group: β1-AS-ODN group, β1-IN-ODN group, 2KIC group, sham operation group, SD group. At 4 weeks after the operation, the mixture of cationic liposomes / β1-ODN (molar ratio 2.0) was injected into the caudal vein of rats. The injection dose was 0.5 mg · kg -1. The blood pressure was measured by Tail-cuff method. Semiquantitative RT The β1 mRNA level was determined by PCR, the receptor density was determined by radioimmunoassay, and the protein level was determined by Western blot. Data were analyzed by one-way ANOVA, with pairwise comparisons using the Newman-Keuls method. Results Compared with 2KIC group, β1-AS-ODN could decrease the blood pressure for 4 weeks (P <0.05) and decrease the maximum blood pressure by 39 mmHg. There was no significant difference in β1-ASN between β1-AS-ODN group and hypertension group (P <0.05). Compared with 2K1C group, the level of β1 receptor in antisense group decreased significantly at 2, 7 and 30 days after injection, which were 29.5%, 35.5% and 20.4 % (P <0.01, P <0.01, P <0.05). There was no significant difference in the level of β2 receptor among the three groups. Conclusion The antisense gene targeting β1 receptor has a significant antihypertensive effect. Β1 antisense oligonucleotide acts after transcription and inhibits the expression of the target gene. The antihypertensive effect of β1 antisense oligonucleotide is significantly different from that of β1 antisense oligonucleotide And the heart β1 receptor binding, but not with β2 receptors.