Abstract
Background To investigate the role of DNA methylation in the control of expression of beta1-AR gene. Detection the expression of beta1-adrenergic receptor (beta1-AR) between different individuals in spontaneously hypertensive rats (SHR) whose genotype is consistent. Methods Measured forty spontaneously hypertensive rats (SHRs) blood pressure (BP) at 14:00-16:00 every day. Using gene sequencing to check SHR beta1-Adrenergic Receptor (beta1-AR) genotype; Real-time quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR), real-time reverse transcriptase-polymerase chain reaction (RT-PCR)to analyze myocardial mRNA expression of beta1-AR gene; methylation specific PCR (MSP) and bisulfite sequencing to detect methylated status in beta1-AR gene; protein expression of beta1-AR were identified by Western blot. Randomly divided them into 176–185mmHg, 186–195mmHg, 196–205mmHg three groups according their BP. Results There was no difference in any two groups in heart rate, body length, tail length, body weight, heart weight (P > 0.05). No mutation was detected in SHR beta1-AR gene sequence; MSP and bisulfite sequencing showed that SHR beta1-AR gene have DNA methylation. Expression of myocardial beta1-AR gene mRNA in methylation and non-methylation group are 27.8 ± 21.7, 110.2 ± 87.5 respectively (P < 0.05). Expression of myocardial beta1-AR gene in both group are 0.2 ± 0.1, 0.4 ± 0.1 respectively (P < 0.05). RT-PCR shows expression of myocardial beta1-AR mRNA are 0.21 ± 0.04, 0.27 ± 0.02, 0.33 ± 0.03 respectively, RQ-PCR shows expression of myocardial beta1-AR mRNA are 25.6 ± 6.4, 67.0 ± 15.5,149.3 ± 10.6 respectively;Expression of myocardial beta1-AR gene are 0.25 ± 0.08, 0.33 ± 0.09, 0.41 ± 0.06 respectively, demonstrates that different tissues of SHR cardiac beta1-AR expression was different, the difference was significant (P < 0.05). Conclusion Beta1-AR gene promoter methylatied in SHR myocardium, and influence the expression of beta1-AR gene. SHR beta1-AR expressed differently, the expression tended to increase with the increase of BP.
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