Estrogen receptors in atherosclerotic human aorta: Inhibition of human vascular smooth muscle cell proliferation by estrogens

Estrogen has been postulated to exert direct anti-atherogenic effects via binding to estrogen receptors (ERs) in vascular smooth muscle cells (VSMCs). Therefore, we believe it is important to examine the status of ER expression in the human cardiovascular system and its disorders. In this study, we first evaluated the relative abundance of messenger RNA (mRNA) of both ER subtypes (ERα and ERβ) in the human aorta using reverse transcription followed by quantitative polymerase chain reaction (RT-qPCR). We then examined the immunolocalization of both ERs in VSMCs of human atherosclerotic lesions. In order to examine which ER subtype was associated with the anti-atherogenic effects of estrogen, we examined the effects of estrogen in two VSMC cell lines, one positive only for ERα and the other positive only for ERβ. The relative abundance of mRNAs for both ERs was higher in female aorta with a mild degree of atherosclerosis than in female aorta with a severe degree of atherosclerosis (P<0.05). In addition, the number of ERα and/or ERβ double positive cells in the neointima was higher in female aorta with a mild degree of atherosclerosis than in female aorta with severe atherosclerosis (P<0.05). Our in vitro study found that estradiol was able to significantly inhibit the proliferation of ERα positive VSMCs but not ERβ positive VSMCs (P<0.05). Moreover, estradiol was found to significantly suppress proliferating cell nuclear antigen (PCNA) mRNA levels in ERα positive VSMCs compared to that of ERβ positive VSMCs, consistent with the findings of cell proliferation. Results from this study suggest that estrogens can inhibit the proliferation of VSMCs through ERα, especially in pre-menopausal women. Our study also indicates that decreased levels of ER, especially ERα, in the female atherosclerotic neointima may be associated with progression of atherosclerotic changes.