Aging is characterized by systemic inflammatory changes and organ dysfunction. In females, loss of estrogen compounds these changes. Post-menopausal women have an abrupt acceleration of atherosclerosis. It would seem that restoration of estrogen would be protective;however, double- blind clinical studies on the use of estrogen replacement have not shown a benefit. Epoxyecosatrienoic acids (EETs), which are lipid signaling molecules, have important effects on angiogenesis, inflammation and are protective against ischemic injury. EETs represent the third pathway of arachidonic acid (AA) metabolism, in addition to prostaglandins and leukotrienes. Given their properties, EETs have the potential to be protective post-menopause. Soluble epoxide hydrolase (sEH) converts EETs to dihydroxy-eicosatrienoic acids (DHETs), which are thought to be pro- inflammatory and to lack the beneficial properties of EETs. In pilot studies we found that vascular sEH increased with aging and that left ventricular sEH expression decreased with estrogen replacement. Estrogen loss combined with aging leads to increased oxidative stress, increased inflammation, dysfunctional EPCs leading to impaired vascular repair, increased inflammation and increased monocyte adhesion. Our hypothesis is that EETs treatment and/or sEH inhibition, which will block hydrolysis of EETs to DHETs, can mitigate the inflammatory changes associated with estrogen loss and aging. We will address this hypothesis with three specific aims: SA 1 - Define the effect of aging vs. estrogen loss on EET metabolism, and the effect of changes in EET/DHETs on vascular function and inflammation. The planned work will investigate changes in EETs with aging and changes in estrogen status. End points will include the effect of EETs vs. estrogen on vascular function, regulation of EETs expression, and the metabolic profile of AA/EETs in aged and adult Norway Brown (NB) rats with and without estrogen. SA2 - Determine the effect of changes in EETs/DHETs vs. aging and E2 loss on EPCs function and proliferation. EETs have vasodilator, angiogenic and anti-inflammatory properties, but their effect on EPCs is not known. We hypothesize that aged EPCs will have impaired function, and that this will be improved by treatment with EETs. Endpoints in the planned work will include EPC proliferation, tube formation and both in vitro and in vivo models of homing and wound repair. SA3. - Investigate the effect of aging vs. estrogen withdrawal and EETs on monocyte activation and adhesion - These experiments explore the effect of aging vs. estrogen on monocyte activation, and how EETs/DHETs modify monocyte activation and adhesion. The planned studies will provide new knowledge of the effect of aging vs. estrogen loss, and the potential role of EETs in ameliorating the inflammatory changes of aging.

Public Health Relevance

The proposed work will investigate the role of EETs, a set of molecules whose synthesis in the body may vary over time, in modulating inflammation and changes in the blood vessels with aging. The research investigates links between changes in estrogen and EETs. An increasing number of women are serving in the armed forces. There are 1.8 million female veterans in the US, which is 7.7% of the veteran population.1 In 2008 the VA provided health care to 281,000 women, an increase of 12% over 2006. It is projected within 25 years 17% of veterans will be female. The median age of female veterans is 47, and as menopause occurs at a mean age of 55, there are a significant number of female veterans who are peri- or post-menopausal. The planned work is very relevant to the veterans'population, which has a high incidence of vascular disease as well as heart disease. Although the work focuses on estrogen and EETs, it will provide new insights and potentially new therapies (sEH inhibitors) vascular disease, which can help both male and female veterans.

National Institute of Health (NIH)
Veterans Affairs (VA)
Non-HHS Research Projects (I01)
Project #
Application #
Study Section
Cardiovascular Studies A (CARA)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
VA Northern California Health Care System
United States
Zip Code
Poe, A J; Knowlton, A A (2017) Exosomes as agents of change in the cardiovascular system. J Mol Cell Cardiol 111:40-50
Sun, Chongxiu; Simon, Scott I; Foster, Greg A et al. (2016) 11,12-Epoxyecosatrienoic acids mitigate endothelial dysfunction associated with estrogen loss and aging: Role of membrane depolarization. J Mol Cell Cardiol 94:180-188
Myers, Richard; Timofeyev, Valeriy; Li, Ning et al. (2015) Feedback mechanisms for cardiac-specific microRNAs and cAMP signaling in electrical remodeling. Circ Arrhythm Electrophysiol 8:942-50
Knowlton, A A; Korzick, D H (2014) Estrogen and the female heart. Mol Cell Endocrinol 389:31-9
Knowlton, Anne A (2012) Estrogen and cardiovascular disease: aging and estrogen loss at the heart of the matter? Future Cardiol 8:9-12
Knowlton, A A; Lee, A R (2012) Estrogen and the cardiovascular system. Pharmacol Ther 135:54-70
Stice, James P; Eiserich, Jason P; Knowlton, A A (2009) Role of aging versus the loss of estrogens in the reduction in vascular function in female rats. Endocrinology 150:212-9