[靶向血管紧张素II 1型受体的短发夹RNA对自发性高血压大鼠血压及AT1R mRNA表达的影响]

[The effects of shRNA targeting angiotensin II type 1 receptor on blood pressure and AT1R mRNA expression in spontaneously hypertensive rats].

作者信息

Xiao Chuan-Shi, Zhang Jin-Lian, Qiu Ling

机构信息

Department of Cardiology, Second Hospital of Shan Xi Medical University, Taiyuan 030001, China.

出版信息

Zhonghua Xin Xue Guan Bing Za Zhi. 2007 Apr;35(4):354-8.

PMID:17711664

Abstract

OBJECTIVE

To investigate the effects of retroviral vector containing shRNA targeting rat angiotensin II type 1 receptor (AT1R) gene (Ad5-AT1R-shRNA) on blood pressure and AT1R mRNA expression in spontaneously hypertensive rats (SHR).

METHODS

Retroviral vector containing shRNA targeting rat AT1R gene was constructed and propagated further in 293 cells. SHR rats were randomly divided into SHR + Ad5-AT1R-shRNA (1.7 x 10(9) TCID(50)/ml) group and SHR (Ad5-EGFP, 7.9 x 10(9) TCID(50)/ml, n = 11 each) and 11 male Wistar-Kyoto rats (WKY) serve as normal controls (Ad5-EGFP, 7.9 x 10(9) TCID(50)/ml). Systolic blood pressure was measured before and after single intravenous injection of Ad5-AT1R-shRNA or Ad5-EGFP. Heart, liver, kidney, aorta and adrenal gland were removed after blood pressure measurement. Tissue Ad5-AT1R-shRNA expression was detected with fluorescence microscope and AT1R mRNA in liver, kidney and aorta was measured by fluorescence quantitative PCR.

RESULTS

Ad5-AT1R-shRNA significantly reduced blood pressure compared with controls (-29 mm Hg, 1 mm Hg = 0.133 kPa, P < 0.05) 24 hours after single injection and this antihypertensive effect could last for 5 to 7 days. Ad5-AT1R-shRNA expression detected with fluorescence microscope was significantly increased in heart, liver, kidney, aorta and adrenal gland post Ad5-AT1R-shRNA injection. AT1R mRNA in kidney and aorta (0.086 +/- 0.014, 0.051 +/- 0.023) were significantly decreased in Ad5-AT1R-shRNA treated rats compared with SHR control rats (0.362 +/- 0.042, 0.463 +/- 0.045, P < 0.01).

CONCLUSION

The results indicate that Ad5-AT1R-shRNA could inhibit the tissue AT1R mRNA expression and produce prolonged antihypertensive effects in SHR rats.

摘要

目的

研究携带靶向大鼠血管紧张素II 1型受体（AT1R）基因的短发夹RNA的逆转录病毒载体（Ad5-AT1R-shRNA）对自发性高血压大鼠（SHR）血压及AT1R mRNA表达的影响。

方法

构建携带靶向大鼠AT1R基因短发夹RNA的逆转录病毒载体，并在293细胞中进一步扩增。将SHR大鼠随机分为SHR + Ad5-AT1R-shRNA（1.7 x 10(9) TCID(50)/ml）组和SHR（Ad5-EGFP，7.9 x 10(9) TCID(50)/ml，每组n = 11），11只雄性Wistar-Kyoto大鼠（WKY）作为正常对照（Ad5-EGFP，7.9 x 10(9) TCID(50)/ml）。单次静脉注射Ad5-AT1R-shRNA或Ad5-EGFP前后测量收缩压。血压测量后取出心脏、肝脏、肾脏、主动脉和肾上腺。用荧光显微镜检测组织中Ad5-AT1R-shRNA的表达，并用荧光定量PCR检测肝脏、肾脏和主动脉中AT1R mRNA的表达。

结果

单次注射后24小时，与对照组相比，Ad5-AT1R-shRNA显著降低血压（-29 mmHg，1 mmHg = 0.133 kPa，P < 0.05），且这种降压作用可持续5至7天。Ad5-AT1R-shRNA注射后，用荧光显微镜检测发现心脏、肝脏、肾脏、主动脉和肾上腺中Ad5-AT1R-shRNA的表达显著增加。与SHR对照大鼠（0.362 +/- 0.042，0.463 +/- 0.045，P < 0.01）相比，Ad5-AT1R-shRNA处理的大鼠肾脏和主动脉中的AT1R mRNA（0.086 +/- 0.014，0.051 +/- 0.023）显著降低。