title: Methylation of adrenergic β1 receptor is a potential epigenetic mechanism controlling antihypertensive response to metoprolol.
abstract: Although metoprolol is used to treat hypertension, clinical responses are variable and unpredictable. Evidence suggests that adrenergic beta1 receptor (ADRB1, designated Adrb1 in rodents) gene polymorphisms influence the level of blood pressure response to this drug therapy, but their presence can not predict the response of the individual patient. The question exists whether epigenetic modifications, such as DNA methylation could cause changes in the gene's expression that are a determining factor in metoprolol's efficacy. The aim of this study was to verify whether DNA methylation could change the expression of the ADRB1 gene, and epigenetic modification could explain why individuals with identical ADRB1 gene polymorphisms have different antihypertensive responses to metoprolol. H9c2 rat myocardial cells in vitro were randomly divided into 5-aza-2'-deoxycytidine (decitabine)-treated (0.5 to 10.0 microM) and control groups. For the in vivo experiments, 45 spontaneously hypertensive rats (SHRs) were divided into metoprolol-treated and control groups, and after a 4-week intervention myocardia were harvested. Genomic methylation-sensitive PCR was used to assess the methylation status of the Adrb1 promoter after DNA extraction from H9c2 cells and SHR myocardia. Real-time fluorescent quantitative RT-PCR was used to determine levels ofAdrb1 mRNA. In H9c2 cells, the least degree of methylation was observed in the 5.0 microM decitabine treated group. Prolonged exposure of cells to 5.0 microM decitabine resulted in downregulating methylation of the Adrb1 promoter. Increased levels of Adrb1 mRNA of the 5.0 microM group demonstrated that this concentration resulted in the highest expression. Accordingly, DNA methylation resulted in the downregulation of Adrb1 transcription. In vivo, the lower level of methylation of the Adrb1 promoter from SHR myocardial samples demonstrated a better antihypertensive effect by metoprolol. The expression of Adrb1 mRNA in the effective group of SHRs was significantly upregulated. In conclusion, as shown in both H9c2 cells and SHRs, downregulated methylation of the Adrb1 promoter is likely to improve the antihypertensive efficacy of metoprolol.
authors.affiliation: Center of Clinical Pharmacology, Third Xiangya Hospital, Central South University, Changsha 410013, China.
authors.forename: Qixia
authors.initials: Q
authors.lastname: Jiang
authors.forename: Hong
authors.initials: H
authors.lastname: Yuan
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Xing
authors.forename: Jingjing
authors.initials: J
authors.lastname: Liu
authors.forename: Zhijun
authors.initials: Z
authors.lastname: Huang
authors.forename: Xia
authors.initials: X
authors.lastname: Du
mesh_terms: D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology*; D018929:Cell Culture Techniques; D019175:DNA Methylation; D044127:Epigenesis, Genetic / Q000235:genetics*; D005260:Female; D006801:Humans; D008790:Metoprolol / Q000494:pharmacology*; D009206:Myocardium / Q000378:metabolism; D011897:Random Allocation; D051381:Rats; D011918:Rats, Inbred SHR; D011941:Receptors, Adrenergic / Q000235:genetics / Q000378:metabolism; D018342:Receptors, Adrenergic, beta-1 / Q000235:genetics* / Q000378:metabolism
chemical_list: D000959:Antihypertensive Agents; D011941:Receptors, Adrenergic; D018342:Receptors, Adrenergic, beta-1; D008790:Metoprolol