Author + information
- Wen Gaiyan and
- Hong Yuan
To explore the effect of downregulation of β1-AR in myocardium of SHRs on antihypertensive effect.
The recombinant plasmid pAAV-ZsGreen-ADRB1-shRNA was confirmed by enzyme digestion and gene sequencing. The plasmid was then packaged to recombinant adeno-associated virus named AAV9- shRNA-ADRB1-ZsGreen. The purity, titer and infection activity in vitro were detected. 18 SHRs were randomly divided into three groups as follows: AAV9-shRNA-ADRB1-ZsGreen injection group, AAV9-CMV-ADRB1-ZsGreen injection group and sham group. SHR myocardium was transfected by injecting virus vector to the pericardial cavity. Rats were killed after 4 weeks. The blood pressure was monitored weekly. The expression of β1-AR in myocardial tissue was assessed by Real-time PCR and Western blot.
The pAAV-ZsGreen-ADRB1-shRNA plasmid was confirmed to be correct by enzyme digestion and gene sequencing. The purified AAV9- shRNA-ADRB1-ZsGreen titer was 1.5x1012vg/ml. In addition, the transfection efficiency of this viral vector to 293 cells reached up to 95%.
β1-AR mRNA was significantly decreased by AAV9-shRNA-ADRB1-ZsGreen injection compared with the negative control group (0.4142 ± 0.3399 vs. 1.0933 ± 0.5853, P < 0.05). The β1-AR protein expression was significantly lower than the negative control group (0.5156 ± 0.1199 vs. 0.8333 ± 0.1728, P < 0.05). Immunohistochemistry results showed that β1-AR protein expression in AAV9-shRNA-ADRB1-ZsGreen injection group was significantly decreased compared with the negative control group and sham group (50.02 ± 4.44 vs. 80.05 ± 3.72 vs. 79.30 ± 2.79, P<0.05).
β1-AR expression in myocardium of SHRs was reduced by AAV9-shRNA-ADRB1-ZsGreen injection. Downregulation of β1-AR in myocardium of SHRs resulted in lower blood pressure.