Demethylation of Circulating Estrogen Receptor Alpha Gene in Cerebral Ischemic Stroke

PLoS One. 2015 Sep 30;10(9):e0139608. doi: 10.1371/journal.pone.0139608. eCollection 2015.


Background: Estrogen is involved in neuron plasticity and can promote neuronal survival in stroke. Its actions are mostly exerted via estrogen receptor alpha (ERα). Previous animal studies have shown that ERα is upregulated by DNA demethylation following ischemic injury. This study investigated the methylation levels in the ERα promoter in the peripheral blood of ischemic stroke patients.

Methods: The study included 201 ischemic stroke patients, and 217 age- and sex-comparable healthy controls. The quantitative methylation level in the 14 CpG sites of the ERα promoter was measured by pyrosequencing in each participant. Multivariate regression model was used to adjust for stroke traditional risk factors. Stroke subtypes and sex-specific analysis were also conducted.

Results: The results demonstrated that the stroke cases had a lower ERα methylation level than controls in all 14 CpG sites, and site 13 and site 14 had significant adjusted p-values of 0.035 and 0.026, respectively. Stroke subtypes analysis showed that large-artery atherosclerosis and cardio-embolic subtypes had significantly lower methylation levels than the healthy controls at CpG site 5, site 9, site 12, site 13 and site 14 with adjusted p = 0.039, 0.009, 0.025, 0.046 and 0.027 respectively. However, the methylation level for the patients with small vessel subtype was not significant. We combined the methylation data from the above five sites for further sex-specific analysis. The results showed that the significant association only existed in women (adjusted p = 0.011), but not in men (adjusted p = 0.300).

Conclusions: Female stroke cases have lower ERα methylation levels than those in the controls, especially in large-artery and cardio-embolic stroke subtypes. The study implies that women suffering from ischemic stroke of specific subtype may undergo different protective mechanisms to reduce the brain injury.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • CpG Islands
  • DNA Methylation*
  • Estrogen Receptor alpha / genetics*
  • Estrogen Receptor alpha / metabolism
  • Female
  • Humans
  • Male
  • Middle Aged
  • Sex Factors
  • Stroke / genetics*
  • Stroke / metabolism*


  • ESR1 protein, human
  • Estrogen Receptor alpha

Grant support

The work was supported by the following: Ministry of Science and Technology (Taiwan, R.O.C. 101-2628-B-037-003-MY2, and 103-2314-B-075A-003-MY3), SHJ; National Health Research Institutes (Taiwan, R.O.C. NHRI-Ex101-10107PI), SHJ; Academia Sinica (BM 104010092), SHJ; Kaohsiung Medical University Hospital (KMUH100-0R47), HFL; and Kaohsiung Medical University (Aim for the Top 500 Universities grant KMU-DT103003, KMU-TP103C003), SHJ.