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KMID : 0390320070170020126
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Chungbuk Medical Journal
2007 Volume.17 No. 2 p.126 ~ p.133
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Role oh High Glucose in the Proliferation and Apoptosis of Endothelial Progenitor Cells
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Hwang Kyung-Kuk
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Abstract
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Purpose: Hyperglycemia plays an important role in the endothelial dysfunction and impaired healing, especially related with diabetes. However its role on the endothelial progenitor cell (EPC), one of the most important cell sources for cell therapy, is rarely investigated. We examined the effect and mechanism of high-glucose on the EPC’s function including the proliferation and survival.
Material and Methods: EPCs were isolated from human peripheral blood by density-gradient centrifugation, and cultured with EGM-2 MV media. EPCs (5× 106 cells/ plate for proliferation, 1.5× 107 cells/plate for apoptosis) were incubated till 48 hours under the 3 different conditions, control (normal glucose level: 5.5 mM/L), high osmolar condition with mannitol (mannitol: 30 mM/L) as a osmotic control for high glucose, and the high glucose (glucose: 30 mM/L). Apoptosis was quantitated by FACS analysis. The activities of proliferation (under the EGM-2 media) and survival (under the serum-free EBM-2 media) of EPCs were evaluated by WST-1 assay (Roche molecular biochemicals,). And, survival-signal pathway were evaluated by Western blot analysis.
Results: Apoptotic cells under the exposure of high glucose were increased, compared to those under the high osmolar condition with mannitol as a osmotic control (48.6 vs 44.5%). The activities of proliferation and survival of EPC was decreased under the high glucose condition. The expression of phospho-Akt and survivin were decreased under the high glucose condition.
Conclusion: This study has the limitation of in-vitro pilot study. These findings suggest that high glucose ameliorate the proliferative and survival capacities of EPC and associate with Akt-Survivin pathway at least in part.
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KEYWORD
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High glucose, Endothelial progenitor cells, Apoptosis, Proliferation, Art
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