Combined oral contraceptive synergistically activates mineralocorticoid receptor through histone code modifications.
Clinical studies have shown that the use of combined oral contraceptive in pre-menopausal women is associated with fluid retention. However, the molecular mechanism is still elusive. We hypothesized that combined oral contraceptive (COC) ethinyl estradiol (EE) and norgestrel (N) synergistically activates mineralocorticoid receptor (MR) through histone code modifications. Twelve-week-old female Sprague-Dawley rats were treated with olive oil (control), a combination of 0.1µg EE and 1.0µg N (low COC) or 1.0µg EE and 10.0µg N (high COC) as well as 0.1 or 1.0µg EE and 1.0 or 10.0µg N daily for 6 weeks. Expression of MR target genes in kidney cortex was determined by quantitative real-time polymerase chain reaction. MR was quantified by western blot. Recruitment of MR and RNA polymerase II (Pol II) on promoters of target genes as well as histone code modifications was analyzed by chromatin immunoprecipitation assay. Treatment with COC increased renal cortical expression of MR target genes such as serum and glucocorticoid-regulated kinase 1 (Sgk-1), glucocorticoid-induced leucine zipper (Gilz), epithelial Na(+)channel (Enac) and Na(+)-K(+)-ATPase subunit α1 (Atp1a1). Although COC increased neither serum aldosterone nor MR expression in kidney cortex, it increased recruitment of MR and Pol II in parallel with increased H3Ac and H3K4me3 on the promoter regions of MR target genes. However, treatment with EE or N alone did not affect renal cortical expression of Sgk-1, Gilz, Enac or Atp1a1. These results indicate that COC synergistically activates MR through histone code modifications.
Department of Pharmacology, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; Cardiovascular Research Institute, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea.
Adedoyin
A
Igunnu
Cardiovascular Research Institute, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; Korea Promotion Institute for Traditional Medicine Industry, Gyeongsan, Gyeongbuk 712-260, Republic of Korea.
Young-Mi
YM
Seok
Department of Pharmacology, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; Cardiovascular Research Institute, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea.
Lawrence A
LA
Olatunji
Department of Pharmacology, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; Cardiovascular Research Institute, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; Cell and Matrix Research Institute, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea.
Seol-Hee
SH
Kang
Department of Pharmacology, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; Cardiovascular Research Institute, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; Cell and Matrix Research Institute, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, Kyungpook National University School of Medicine, Daegu 700-842, Republic of Korea. Electronic address: inkim@knu.ac.kr.
Inkyeom
I
Kim
D000818:Animals; D001483:Base Sequence; D003277:Contraceptives, Oral, Combined / Q000494:pharmacology*; D004357:Drug Synergism; D004997:Ethinyl Estradiol / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D042421:Histone Code / Q000187:drug effects*; D017874:Immediate-Early Proteins / Q000235:genetics; D007672:Kidney Cortex / Q000187:drug effects / Q000378:metabolism; D009644:Norgestrel / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000187:drug effects; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D021381:Protein Transport / Q000187:drug effects; D012319:RNA Polymerase II / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018161:Receptors, Mineralocorticoid / Q000737:chemistry* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics; D000099184:Serum-Glucocorticoid Regulated Kinases
D003277:Contraceptives, Oral, Combined; D004997:Ethinyl Estradiol; D017874:Immediate-Early Proteins; D009644:Norgestrel; D017346:Protein Serine-Threonine Kinases; D012319:RNA Polymerase II; D018161:Receptors, Mineralocorticoid; D014157:Transcription Factors; D000099184:Serum-Glucocorticoid Regulated Kinases; C498609:Tsc22d3 protein, rat
Ethinyl estradiol (PubChem CID: 5991) and norgestrel (PubChem CID: 5702096).; Gene transcription; Histone code; Mineralocorticoid receptor; Oral contraceptive