Prenatal caffeine ingestion induces aberrant DNA methylation and histone acetylation of steroidogenic factor 1 and inhibits fetal adrenal steroidogenesis.
Prenatal caffeine ingestion is one of the risk factors for intrauterine growth retardation (IUGR). Adrenal plays a pivotal role, mainly through steroidogenesis, in the regulation of intrauterine homeostasis and in fetal development and maturation. We have shown that prenatal caffeine ingestion can inhibit fetal adrenal corticosterone production, but the underlying mechanism is unknown. This study investigated the effects of prenatal caffeine ingestion on corticosterone and its associated synthesized enzymes (steroidogenic acute regulatory protein, StAR; 3β-hydroxysteroid dehydrogenase, 3β-HSD; cytochrome P450 cholesterol side chain cleavage, P450scc; P450c21; and P450c11) in the fetal adrenal in rats and further explored the underlying mechanism by analyzing the epigenetic modification and expression of the key transcription factor steroidogenic factor-1 (SF-1). The pregnant rats were intragastrically treated with 120 mg/kg.d caffeine from gestational day 11-20. The results showed that the IUGR rate was 51.2% after caffeine treatment. The contents of corticosterone and the mRNA levels of StAR, P450scc, P450c21, and P450c11 were decreased significantly in the fetal adrenal. Furthermore, caffeine reduced both the protein and the mRNA expression of SF-1 in the fetal adrenal. The epigenetic analysis showed that caffeine treatment can significantly enhance the mRNA expression of DNA methyltransferase (Dnmt) 1, Dnmt3a, histone deacetylases (Hdac) 1, and Hdac2. The detection of DNA methylation by bisulfite-sequencing PCR uncovered a notably increased total methylation rate in the SF-1 promoter. The ChIP assay showed decreased acetylation levels of H3K9 and H3K14 in the SF-1 promoter. In conclusion, prenatal caffeine ingestion is able to induce aberrant DNA methylation and histone acetylation of the SF-1 promoter in the rat fetal adrenal. These effects may contribute to the inhibition of the expression of SF-1 and its associated steroidogenic enzymes and the production of corticosterone during fetal development.
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China; Research Center of Food and Drug Evaluation, Wuhan University, Wuhan 430071, China.
Jie
J
Ping
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China.
Jian-fei
JF
Wang
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China; Department of Pharmacology, Medical School of Yangtze University, Jingzhou 434000, China.
Lian
L
Liu
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China.
You-e
YE
Yan
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China.
Fang
F
Liu
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China.
You-ying
YY
Lei
Department of Pharmacology, Basic Medical School of Wuhan University, Wuhan 430071, China; Research Center of Food and Drug Evaluation, Wuhan University, Wuhan 430071, China. Electronic address: wanghui19@whu.edu.cn.
Hui
H
Wang
D000107:Acetylation; D000311:Adrenal Glands / Q000187:drug effects / Q000196:embryology / Q000378:metabolism*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002110:Caffeine / Q000633:toxicity*; D000697:Central Nervous System Stimulants / Q000633:toxicity*; D047369:Chromatin Immunoprecipitation; D003345:Corticosterone / Q000494:pharmacology; D019175:DNA Methylation / Q000187:drug effects*; D044127:Epigenesis, Genetic / Q000187:drug effects; D005260:Female; D006657:Histones / Q000378:metabolism*; D007150:Immunohistochemistry; D011247:Pregnancy; D012313:RNA; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D054339:Steroidogenic Factor 1 / Q000378:metabolism*; D013256:Steroids / Q000096:biosynthesis*
D000893:Anti-Inflammatory Agents; D000697:Central Nervous System Stimulants; D006657:Histones; C034477:RNA primers; D054339:Steroidogenic Factor 1; D013256:Steroids; D002110:Caffeine; D012313:RNA; D003345:Corticosterone
Caffeine; DNA methylation; Fetal adrenal; Histone acetylation; Intrauterine growth retardation (IUGR); Steroidogenic factor-1 (SF-1)