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Disruption to the vascular homoeostasis is detrimental in vascular diseases. This study examined how the combination of homocysteine, adenosine and tumor necrosis factor-alpha (TNF-α) influenced endothelial cell survival. In cultured human-derived cardiovascular (EA.hy926) and cerebrovascular (HBEC-5i) endothelial cells, cell death events were initiated by TNF-α (0.1-10 ng/mL) only when both homocysteine (0.5 mM) and adenosine (0.5 mM) were present. The accelerated cell death events induced by the combination were triggered through excessive apoptosis. This was evident by membrane phospholipid phosphatidylserine externalisation, cell shrinkage and DNA fragmentation, as well as an increase in the expressions and occurrence of active caspase-3 and cleaved poly(ADP-ribose) polymerase (PARP) positive cells. Collectively, homocysteine, adenosine and TNF-α are interrelated in the survival of endothelial cells, and this co-existence should be considered in future drug development for cardiovascular and cerebrovascular diseases.

Combination of TNF-a, Homocysteine and Adenosine Exacerbated Cytotoxicity in Human Cardiovascular and Cerebrovascular Endothelial Cells