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Background Data Discussion Methods Results References Acknowledgements
Exploring Age and Estrogen’s Cardioprotective Effect on Co-Cultured VSMC and Endothelial Cells using Endothelin-1 as a Marker B. Dhesi, H. Hwang, A. Knowlton MD Molecular and Cellular Cardiology, Cardiovascular Division, UC Davis School of Medicine Background Data Discussion There is an increased cardiovascular risk in post menopausal women; estrogen has been proposed to have a cardio protective effect. The mechanism of protection needs further investigation and may provide insight into future cardiovascular treatments. Understanding the effects of Senescent (SEN) vs Early passage (EP) endothelial cells may provide us with information for treatment of cardiovascular complications associated with senescence, specifically hypertension. The goal of this project is to determine the effects of age and presence of estrogen on the expression of Endothelin-1 (ET-1), a potent vasoconstrictor. VSMC co cultured with EC show a decrease in ET-1 expression. The relationship between VSMC and EC may provide a “brake” on ET-1 expression through intracellular messaging. Senescent cells show an increased ET-1 expression than early passage cells, this may explain the increase in blood pressure that may be related to increased age. Estrogen treatment did not decrease ET-1 expression in SEN only culture however it did decrease ET-1 in co cultured VSMC/SEN cells. This indicates that Estrogen may act through VSMC “brake” to reduce ET-1 expression in Endothelial cells. Future bench research can be done to see the presence of Estrogen receptor on VSMC and look for markers that change on VSMC due to estrogen. Estrogen’s beneficial cardiovascular effects must be weighed against the increased endometrial and breast cancer risk. Future research can evaluate the effects of estrogen replacement therapy on cardiovascular event outcome and cancer risk. ET-1 Assay Methods Vascular smooth muscle cells (VSMC) and endothelial cells (EC) were co cultured together in a well separated by a thin microporous membrane. ET-1 was then assayed using ELISA (Endothelin-1 Quantikine ELISA kit) on the media. Estrogen treatment was compared to Vehicle (DMSO). Endothelial cells were from human coronary artery endothelial from female donors (age 17-63), VSMC were obtained from human coronary artery vascular smooth muscle cells from a 43 YO female. Figure 2: Endothelin-1 Assay Results. Data was analyzed with SigmaPlot using 2-way ANOVA Results References VSMC do not produce ET-1. When VSMC are cultured with Early Passage cells there is a reduction in ET-1 expression when compared to Early Passage alone. In vehicle treated co-culture experiments, co cultured senescent cells show a greater ET-1 expression than co cultured early passage cells. Estrogen treated VSMC/SEN show a decrease in ET-1 expression compared to Vehicle VSMC/SEN. K.L. Hamilton, F.N. Mbai, S. Gupta, A.A. Knowlton, Estrogen, Heat Shock Proteins, and NFB in Human Vascular Endothelium, Arterioscler Thromb Vasc Biol. 2004;24: A.A. Knowlton, D.H. Korzick, Estrogen and the female heart, Molecular and Cellular Endocrinology. 2014;389(1-2):31-39. Acknowledgements I would like to thank UC Davis School of Medicine for giving me the opportunity to engage in research through the MSRF program. I would also like to give my deepest gratitude to Dr. Knowlton and Hyun Hwang for mentoring me throughout this project. This project was supported by a VA Merit Award from Sacramento VA. Figure 1: Diagram of culturing methods.
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