摘要
目的用RNA干扰(RNAi)技术同时下调血管紧张素转换酶(ACE)和血管紧张素Ⅱ1型受体(AT_1R),观察其对自发性高血压大鼠(SHR)血压及主动脉重构的影响。方法 30只SHR随机分为5组,每组6只:空白对照组(注射生理盐水);病毒对照组(注射对照腺病毒);重组腺病毒载体(Ad5)-ACE-短发夹环RNA(shRNA)治疗组;Ad5-AT_1R-shRNA治疗组;Ad5-ACE-AT_1R-shRNA治疗组(分别注射表达ACE基因、AT_1R基因、ACE和AT_1R基因特异shRNA的重组腺病毒载体);同时设WKY正常血压对照组(注射生理盐水)。以上各组大鼠均于实验的第1、17天各注射1次,干预前后检测血压、心率的变化。于首次注射后第3天,用实时荧光定量聚合酶链式反应(PCR)检测主动脉ACE和AT_1R mRNA的表达;用Western blot检测主动脉ACE和AT_1R蛋白的表达。实验结束时做主动脉组织光镜病理检查观察主动脉结构。结果首次注射后第3天,Ad5-ACE-shRNA、Ad5-AT_1R-shRNA、Ad5-ACE-AT_1R-shRNA治疗组尾动脉压分别下降(25.2±5.3)、(23.5±4.8)、(27.1±6.4)mm Hg,3个治疗组降压效应可持续15 d,SHR空白对照组和病毒对照组尾动脉压持续升高,正常血压对照组尾动脉压无明显变化,治疗组较空白对照组和病毒对照组下降(均P<0.05)。Ad5-ACE-shRNA、Ad5-AT_1R-shRNA、Ad5-ACE-AT_1R-shRNA治疗组主动脉组织中的ACE、AT_1R mRNA和蛋白表达明显低于空白对照组、病毒对照组(均P<0.05)。光镜观察显示3个治疗组主动脉结构明显改善,Ad5-ACE-AT_1R-shRNA治疗组最显著。结论 ACE和AT_1R双基因沉默,产生较单基因位点的阻断更明显的降压及逆转主动脉重构效应。
Objective To observe the effects of simultaneously down-regulate angiotensin converting enzyme(ACE) and angiotensin Ⅱ type 1 receptor(AT1R) by RNA interference(RNAi) technique on blood pressure and aortic remodeling in spontaneously hypertensive rats(SHR). Methods Thirty SHR were randomly divided into control group(i.v. injection of saline, n=6), virus control group(injection of adenovirus, n=6), Ad5-ACE-shRNA [i.v. injection of recombinant adenovirus vector(Ad5)-ACE-short hairpin RNA(shRNA), n=6], AT1R-shRNA treatment group(n=6), and Ad5-ACE-AT1R-shRNA treatment group(n=6);WHY rats were served as normal blood pressure control group(injection of saline, n=6). The changes of blood pressure(BP) and heart rate(HR) were measured before and after miRNA or saline injected at the 1 st and 17 th day of the experiment. The expression of mRNA and protein of ACE and AT1R in the aorta were detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR) and Western blot on the third day after the first injection. The aortic structure of aortic tissue was examined by light microscopy at the end of the experiment. Results The BP of Ad5-ACE-shRNA group, Ad5-AT1R-shRNA group and Ad5-ACE-AT1R-shRNA group decreased respectively by(25.2±5.3),(23.5±4.8) and(27.1±6.4) mm Hg compared with control group or virus control group(P<0.05). The antihypertensive effects in Ad5-ACE-shRNA group, Ad5-AT1R-shRNA group and Ad5-ACE-AT1R-shRNA group continued for 15 days. Conversely, the BP in SHR blank control group or SHR virus control group increased significantly, though the BP in WKY control group didn’t alter significantly. Compared with blank control group and virus control group, the mRNA, protein expression of ACE and AT1R in Ad5-ACE-shRNA, Ad5-AT1R-shRNA and Ad5-ACE-AT1R-shRNA group decreased significantly(P<0.05). Furthermore, it showed that the structure of the aorta was significantly improved in the three treatment groups, especially in Ad5-ACE-AT1R-shRNA treatment group. Conclusion ACE and AT1R double gene silencing produced more obvious antihypertensive effects and reversed more aortic remodeling than single gene silencing.
作者
周华
张翠芳
陈瑞瑞
高奋
何军华
杨志明
肖传实
ZHOU Hua;ZHANG Cui-fang;CHEN Rui-rui;GAO Fen;HE Jun-hua;YANG Zhi-ming;XIAO Chuan-shi(Department o f Cardiology,Second Hospital of Shanxi Medical University,Taiyuan Shanxi 030001,China)
出处
《中华高血压杂志》
CAS
CSCD
北大核心
2019年第2期149-155,共7页
Chinese Journal of Hypertension
