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Proteomics analysis of the proliferative effect of low-dose ouabain on human endothelial cells
Jie Qiu1, Hai-Qing Gao, Rui-Hai Zhou
1Department of Geriatrics, Shandong University Qilu Hospital, Jinan, China.
Abstract:
Digitalis has been used to treat congestive heart failure for more than 200 years, although the dual effects (proliferation and death) induced by digitalis on cell growth have been known for many years, the mechanisms by which digitalis causes the actions were not completely known. The aim of this work was to characterize the proliferative effect of ouabain on cell growth in endothelial cells, and, to do the differential proteomic analysis of human umbilical vein endothelial cells (HUVEC) in response to ouabain and examine changes in protein expression. HUVEC were exposed to different concentrations (0.1-100 nM) of ouabain at 12-48 h intervals. Cell growth and morphological changes of HUVEC treated with ouabain were compared with cells under nontreated conditions. Ouabain stimulated HUVEC cell proliferation at low concentrations and induced cell death at higher concentrations. Using proteomics study, we identified 32 proteins of HUVEC with various important cellular functions and revealed 8 proteins such as Annexin A1, Annexin A2, Malate dehydrogenase, Myosin regulatory light chain 2 (MRLC2), Profilin-1, S100 calcium-binding protein A13, Triosephosphate isomerase and Translationally controlled tumor protein, regulated by low-dose ouabain treatment and MRLC2 was subsequently confirmed by Western blot. Our results give new insights into the cellular and molecular mechanisms of the proliferation action of low-dose ouabain on HUVEC and provide new avenues for the treatment of cardiovascular diseases.
Insights
Low-dose ouabain promotes endothelial cell proliferation, offering new insights into cardiovascular disease treatment. This study identified key proteins, including Myosin regulatory light chain 2 (MRLC2), involved in ouabain
Area of Science:
- Cardiovascular Biology
- Cellular Proteomics
- Pharmacology
Background:
- Digitalis compounds, like ouabain, have a long history in treating heart failure.
- The dual effects of digitalis on cell growth (proliferation and death) are known, but mechanisms remain unclear.
- Understanding ouabain's cellular actions is crucial for developing new cardiovascular therapies.
Purpose of the Study:
- To characterize the proliferative effect of ouabain on endothelial cell growth.
- To perform differential proteomic analysis of human umbilical vein endothelial cells (HUVEC) in response to ouabain.
- To identify specific proteins regulated by low-dose ouabain treatment in HUVEC.
Main Methods:
- HUVEC were treated with varying ouabain concentrations (0.1-100 nM) for 12-48 hours.
- Cell growth and morphology were assessed and compared to control groups.
- Proteomics analysis identified differentially expressed proteins, with MRLC2 validated by Western blot.
Main Results:
- Low-dose ouabain (0.1-100 nM) stimulated HUVEC proliferation.
- High concentrations of ouabain induced cell death.
- Proteomics identified 8 key proteins, including Annexin A1, Annexin A2, and MRLC2, regulated by low-dose ouabain.
Conclusions:
- Low-dose ouabain exhibits a proliferative effect on HUVEC.
- Specific proteins, such as MRLC2, are involved in ouabain-induced endothelial cell proliferation.
- These findings provide novel insights into the molecular mechanisms of ouabain's action and potential therapeutic strategies for cardiovascular diseases.
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