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Influence of rAAV1 Mediated β1-AR Gene Overexpression on Metoprolol Antihypertensive Effect of Spontaneously Hypertensive Rats

Author: ZhangMeiZhen
Tutor: YuanHong
School: Central South University
Course: Pharmacy
Keywords: Recombinant adeno-associated virus vector β1- adrenergic receptor Hypertension Metoprolol
CLC: R544.1
Type: Master's thesis
Year: 2011
Downloads: 25
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Abstract


Objective: To evaluate rAAV1/CMV-Adrb1 increased SHR myocardial tissue β1-AR expression on antihypertensive effect of metoprolol . Method : 1. Using gene synthesis of rat β1-AR gene open reading frame length , constructed pSNAV1/CMV-Adrbl plasmid sequencing using gene recombinant plasmid was identified for the correct identification of pSNAV1/CMV-Adrb1 recombinant adeno-associated virus rAAV1/CMV-Adrb1 packaging. Two experimental subjects were 24 SHR, randomly selected six as rAAV1/CMV-EGFP injection group , and the remaining 18 SHR were randomly divided into three groups : rAAV1/CMV-Adrb1 metoprolol group 6 , sham metoprolol group 6 , sham saline group 6 , metoprolol group received 25mg/kg / gavage and weekly monitoring of blood pressure in SHR level, the rats were sacrificed after 4 weeks of intervention and take the myocardial tissue . 3 using RT-PCR and Real-time RT-PCR assay β1-AR mRNA in myocardium in SHR tissues ; using immunohistochemistry and Western-blot assay β - AR protein content . Results: 1 Construction and identification of viral vectors : gene sequencing confirmed pSNAV1/CMV-Adrb1 successfully constructed , rAAV1/CMV-Adrb1 viral titer of 2.0 × 1012v.g./mL. Two in each group SHRβ1-AR expression comparison : RT-PCR and Real-time RT-PCR results showed that , rAAV1/CMV-Adrb1 after injection β1-AR mRNA expression was significantly higher than the sham group (0.587 ± 0.132 vs.0.212 ± 0.075 , P lt; 0.05); immunohistochemistry and Western-blot showed that , rAAV1/CMV-Adrb1 injection group β1-AR protein expression was significantly higher than the sham group (0.545 ± 0.119 vs.0.421 ± 0.136, P lt; 0.05) . Three in each group SHR antihypertensive efficacy comparison: After 4 weeks , rAAV1/CMV-Adrb1 metoprolol metoprolol group and sham operation group systolic blood pressure was significantly lower than the sham saline group (176.2 ± 8.5mmHg vs. 182.5 ± 4.5mmHg vs.198.7 ± 7.0 mmHg, P lt; 0.05); 4 Chow metoprolol after the intervention , rAAV1/CMV-Adrb1 magnitude injection group was significantly lower systolic blood pressure compared with the sham group (27.8 ± 8.5mmHg vs. 16.8 ± 2.5mmHg, P lt; 0.05). Conclusions : 1. RAAV1/CMV-Adrb1 recombinant adeno-associated virus vector was constructed successfully and significantly increased SHR myocardial tissue β1z-AR expression . 2.β1-AR overexpression SHR better antihypertensive efficacy of metoprolol , may affect the antihypertensive effect of metoprolol one of the factors of individual differences .

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CLC: > Medicine, health > Internal Medicine > Heart, blood vessels ( circulatory ) disease > Abnormal blood pressure > Hypertension
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