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RNA干扰下调ACE和AT1R对自发性高血压大鼠血压及脑组织损伤的影响
作者:
作者单位:

(山西医科大学第二医院心血管内科,山西省太原市 030001)

作者简介:

周华,博士,副主任医师,研究方向为心血管内科,E-mail为zhouhua032670@sina.com。


Effects of RNA interference on blood pressure and brain tissue injury in spontaneously hypertensive rats by downregulating ACE and AT1R
Author:
Affiliation:

Department of Cardiology, Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi, China)

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    摘要:

    目的 用RNA干扰下调血管紧张素转换酶(ACE)和血管紧张素Ⅱ1型受体(AT1R),观察其对自发性高血压大鼠(SHR)血压及脑组织损伤的影响。 方法 SHR随机分为5组:空白对照组、病毒对照组、Ad5-ACE-shRNA治疗组、Ad5-AT1R-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组,同时设WKY正常血压对照组。各组大鼠均于实验第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,酶联免疫吸附法检测大鼠脑组织中ACE、AngⅡ含量,实时荧光定量PCR检测脑组织ACE mRNA、AT1R mRNA表达,Western blot检测脑组织ACE、AT1R蛋白表达。实验结束时做脑组织光镜病理检查观察脑结构。 结果 于首次注射后第3天,Ad5-ACE-shRNA治疗组、Ad5-AT1R-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组尾动脉压分别下降25.2±5.3 mmHg、23.5±4.8 mmHg、27.1±6.4 mmHg,第14天分别下降24.4±5.6 mmHg、23.6±4.9 mmHg、26.3±6.9 mmHg,第17天时血压有所回升；第2次注射后Ad5-ACE-shRNA治疗组、Ad5-AT1R-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组尾动脉压再次明显下降,第23天时较首次注射前分别下降40.2±4.5 mmHg、37.8±3.9 mmHg、43.7±7.3 mmHg,第37天时血压有所回升,第40天时血压上升至第2次注射前水平；空白对照组和病毒对照组血压则持续升高26 mmHg,两组比较差异无统计学意义；正常血压对照组尾动脉压无明显变化。Ad5-ACE-shRNA治疗组、Ad5-AT1R-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组脑组织ACE、AngⅡ显著低于空白对照组和病毒对照组(P＜0.05),而与正常血压对照组比较差异无统计学意义。Ad5-ACE-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组脑组织ACE mRNA和Ad5-AT1R-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组脑组织AT1R mRNA的表达明显低于空白对照组、病毒对照组(P＜0.05),而与正常血压对照组比较差异无统计学意义。Ad5-ACE-shRNA治疗组、Ad5-AT1R-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组脑组织AT1R蛋白表达显著低于空白对照组、病毒对照组(P＜0.05)。光镜显示Ad5-ACE-shRNA治疗组、Ad5-AT1R-shRNA治疗组、Ad5-ACE-AT1R-shRNA治疗组脑组织结构明显改善。 结论 RNA干扰下调ACE和AT1R表达,降低SHR血压并改善脑损伤,尤以Ad5-ACE-AT1R-shRNA治疗组效果明显且副作用少。RNA干扰对高血压的基因治疗具有良好应用前景。

    Abstract:

    Aim RNA interference was used to downregulate angiotensin converting enzyme (ACE) and angiotensinⅡtype 1 receptor (AT1R), and to observe its effects on blood pressure and brain tissue injury in spontaneously hypertensive rats (SHR). Methods SHR was randomly divided into 5 groups:blank control group, virus control group, Ad5-ACE-shRNA treatment group, Ad5-AT1R-shRNA treatment group, Ad5-ACE-AT1R-shRNA treatment group, and WKY normal blood pressure control group was set up. All rats in each group were injected 1 time in the first and seventeenth day of the experiment. On the third day after the first injection, enzyme-linked immunosorbent assay (ELISA) was used to detect the contents of ACE and Ang Ⅱ in rat brain, real-time fluorescent quantitative PCR was used to detect the expression of ACE mRNA and AT1R mRNA in brain tissue, Western blot was used to detect the expression of ACE and AT1R protein in brain tissue. At the end of the experiment, brain tissue was examined by light microscopy to observe the brain structure. Results On the third day after the first injection, the tail arterial pressure of Ad5-ACE-shRNA treatment group, Ad5-AT1R-shRNA treatment group and Ad5-ACE-AT1R-shRNA treatment group decreased by 25.2±5.3 mmHg, 23.5±4.8 mmHg and 27.1±6.4 mmHg, 24.4±5.6 mmHg, 23.6±4.9 mmHg and 26.3±6.9 mmHg respectively on the 14th day, and the blood pressure rose on the 17th day after Ad5-ACE-shRNA treatment and Ad5-AT1R-shRNA treatment after the second injection The tail arterial pressure of Ad5-ACE-AT1R-shRNA treatment group decreased again. On the 23rd day, the arterial pressure decreased by 40.2±4.5 mmHg, 37.8±3.9 mmHg and 43.7±7.3 mmHg respectively before the first injection. On the 37th day, The blood pressure rose to the level before the second injection on the 40th day. The blood pressure of the blank control group and the virus control group continued to increase by 26 mmHg, there was no significant difference between the two groups; the tail arterial pressure of the normal blood pressure control group had no obvious change. The expression of ACE mRNA in brain tissue of Ad5-ACE-shRNA treatment group and Ad5-ACE-AT1R-shRNA treatment group and the expression of AT1R mRNA in the brain tissue of Ad5-AT1R-shRNA treatment group and Ad5-ACE-AT1R-shRNA treatment group were significantly lower than those of blank control group and virus control group (P＜0.05), but there was no significant difference compared with normal blood pressure control group. The expression of AT1R protein in brain tissue of Ad5-ACE-shRNA treatment group, Ad5-AT1R-shRNA treatment group and Ad5-ACE-AT1R-shRNA treatment group was significantly lower than that of blank control group and virus control group (P＜0.05). The light microscope showed that the brain tissue structure of Ad5-ACE-shRNA treatment group, Ad5-AT1R-shRNA treatment group and Ad5-ACE-AT1R-shRNA treatment group was significantly improved.Conclusion RNA interference downregulated ACE and AT1R expression, reduced blood pressure in spontaneously hypertensive rats and improved brain injury, especially Ad5-ACE-AT1R-shRNA treatment had obvious effect and less side effect. RNA interference has a good application prospect for gene therapy of hypertension.

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    引证文献
引用本文

周华,张翠芳,陈瑞瑞,李瑾,高奋,杨志明. RNA干扰下调ACE和AT1R对自发性高血压大鼠血压及脑组织损伤的影响[J].中国动脉硬化杂志,2018,26(2):157~164.

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  • 收稿日期:2017-07-07
  • 最后修改日期:2017-09-03
  • 在线发布日期: 2018-03-07

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