Estrogen-dependent epigenetic regulation of soluble epoxide hydrolase via DNA methylation

Abstract

To elucidate molecular mechanisms responsible for the sexually dimorphic phenotype of soluble epoxide hydrolase (sEH) expression, we tested the hypothesis that female-specific down-regulation of sEH expression is driven by estrogen-dependent methylation of the gene. Mesenteric arteries isolated from male, female, ovariectomized female (OV), and OV with estrogen replacement (OVE) mice, as well as the human cell line (HEK293T) were used. Methylation-specific PCR and bisulfite genomic sequencing analysis indicate significant increases in DNA/CG methylation in vessels of female and OVE compared with those of male and OV mice. The same increase in CG methylation was also observed in male vessels incubated with a physiological concentration of 17β-estradiol (17β-E) for 48 hours. All vessels that displayed increases in CG methylation were concomitantly associated with decreases in their mRNA and protein, suggesting a methylation-induced gene silencing. Transient transfection assays indicate that the activity of promoter-coding luciferase was significantly attenuated in HEK293T cells treated with 17β-E, which was prevented by additional treatment with an estrogen receptor antagonist (ICI). ChIP analysis indicates significantly reduced binding activities of transcription factors (including SP1, AP-1, and NF-κB with their binding elements located in the promoter) in vessels of female mice and human cells treated with 17β-E, responses that were prevented by ICI and Decitabine (DNA methyltransferase inhibitor), respectively. In conclusion, estrogen/estrogen receptor-dependent methylation of the promoter of gene silences sEH expression, which is involved in specific transcription factor-directed regulatory pathways.

Neonatal gut and immune maturation is determined more by postnatal age than by postconceptional age in moderately preterm pigs

“…As noted above, such effects are unlikely to play a major role for the developmental trajectories observed in this study. Normally, preterm pigs delivered at or slightly before 90% gestation (0.6 -1.1 kg birth wt) show variable degrees of respiratory distress, reduced systemic immunity, and high NEC sensitivity, especially when fed rapidly increasing amounts of infant formulas (46,51,53). These symptoms were generally absent for the present moderately preterm pigs (1.1 kg at birth), in part because we fed gradually increasing volumes of bovine colostrum, which protects preterm pigs against NEC and sepsis (10,49), followed by a slow transition to mature bovine milk and without use of antibiotics.…”

The histone demethylase Jarid1b mediates angiotensin II‐induced endothelial dysfunction by controlling the 3′UTR of soluble epoxide hydrolase

“…Increases in DNA/CG methylation of EPHX2 , a pattern dependent on estrogens and which is found more frequently in females, is associated with reduced promoter activity. Thus, oestrogens decrease sEH mRNA and protein abundance and DNA methyltransferase inhibitors increase its expression . Chromatin immunoprecipitation experiments revealed that binding of the transcription factors SP‐1, AP‐1 and NF‐κB to the promoter of EPHX2 is reduced in female mice and human cells after treatment with 17ß‐estradiol .…”

“…Thus, oestrogens decrease sEH mRNA and protein abundance and DNA methyltransferase inhibitors increase its expression . Chromatin immunoprecipitation experiments revealed that binding of the transcription factors SP‐1, AP‐1 and NF‐κB to the promoter of EPHX2 is reduced in female mice and human cells after treatment with 17ß‐estradiol . The role of histone modifications in this context is, however, unknown so far.…”

Estradiol Metabolism: Crossroads in Pulmonary Arterial Hypertension

“…In contrast, 2ME inhibits CYP1B1 activity and reduces HETE formation and cardiac remodeling in rats with pressure overload-induced cardiac hypertrophy [114]. Moreover, E2 inhibits the expression/activity of soluble epoxide hydrolase (sEH), a key enzyme in EET metabolism [144]. Several studies linked low sEH activity to the pathophysiology of PAH: (1) E2-, genetic-, and pharmacologically-induced downregulation of sEH (and increased pulmonary EET) potentiates hypoxic pulmonary vasoconstriction [145][146][147]; (2) lungs from PH patients express no/little sEH; (3) hypoxia decreases the expression of sEH; and (4) when exposed to hypoxia, sEH KO mice exhibit exacerbated pulmonary vascular remodeling [147].…”