Equilin in conjugated equine estrogen increases monocyte-endothelial adhesion via NF-κB signaling

• Published in: PloS one 2019-01, Vol.14 (1), p.e0211462-e0211462
• Main Authors: Ito, Fumitake, Mori, Taisuke, Tarumi, Yosuke, Okimura, Hiroyuki, Kataoka, Hisashi, Tanaka, Yukiko, Koshiba, Akemi, Kitawaki, Jo
• Format: Article
• Language: English
• Subjects: 17β-Estradiol; Adhesion; Animals; Arteriosclerosis; Atherosclerosis; Biology and Life Sciences; Cardiovascular disease; Cell Adhesion; Cell adhesion & migration; Cell adhesion molecules; Cell Adhesion Molecules - metabolism; Cells, Cultured; Clinical trials; E-selectin; Endothelial cells; Enzyme immunoassay; Equilin - pharmacology; Estrogen receptors; Estrogens; Estrogens, Conjugated (USP) - pharmacology; Estrone; Gene expression; Gynecology; Health risk assessment; Hormone replacement therapy; Horses; Human Umbilical Vein Endothelial Cells - drug effects; Human Umbilical Vein Endothelial Cells - physiology; Humans; Hypoxia; Immunoassay; Intercellular adhesion molecule 1; Kinases; Medicine; Medicine and Health Sciences; Monocytes; Monocytes - drug effects; Monocytes - physiology; mRNA; NF-κB protein; Obstetrics; Polymerase chain reaction; Proteins; Research and Analysis Methods; Sex hormones; Signal Transduction; Studies; Sulfates; Therapy; Umbilical vein; Womens health
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0211462

The adhesion of monocytes to endothelial cells, which is mediated by adhesion molecules, plays a crucial role in the onset of atherosclerosis. Conjugated equine estrogen, which is widely used for estrogen-replacement therapy, contains both estrone sulfate and various nonhuman estrogens, including equilin. To investigate the association between various estrogen types and atherosclerosis risk, we examined their effect on adhesion-molecule expression in human umbilical vein endothelial cells (HUVECs). In estrogen-treated HUVECs, the mRNA and protein expression levels of adhesion molecules were quantified by real-time polymerase chain reaction and enzyme immunoassay. Additionally, a flow-chamber system was used to assess the effects of estrogens on the adherence of U937 monocytoid cells to HUVECs. Equilin, but not 17β-estradiol (E2) or other types of estrogen, significantly increased the mRNA (P < 0.01) and protein (P < 0.05) expression of the adhesion molecules E-selectin and intercellular adhesion molecule-1 as compared with levels in controls. Equilin treatment increased the adherence of U937 monocytoid cells to HUVECs relative to the that in the control (P < 0.05), decreased estrogen receptor (ER)β expression, and increased the expression of proteins involved in nuclear factor kappa-B (NF-κB) activation relative to levels in controls. Furthermore, the accumulation of NF-κB subunit p65 in HUVEC nuclei was promoted by equilin treatment. By contrast, E2 treatment neither increased the number of adhered monocytoid cells to HUVECs nor altered the expression of ERβ or NF-κB-activating proteins. Our findings suggest that in terms of the adhesion of monocytes at the onset of atherosclerosis, E2 may be preferable for estrogen-replacement therapy. Further studies comparing equilin treatment with that of E2 are needed to investigate their differential impacts on atherosclerosis.