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Inhibition of ATP‐induced calcium influx by homocysteine in human umbilical vein endothelial cells
Author(s) -
Cortés Magdalena P.,
Becerra Juan P.,
Vinet Raúl,
Álvarez Rocío,
Quintana Irene
Publication year - 2013
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1002/cbin.10077
Subject(s) - umbilical vein , calcium , homocysteine , nitric oxide , chemistry , endocrinology , medicine , intracellular , calcium in biology , incubation , cytosol , hyperhomocysteinemia , biochemistry , in vitro , enzyme
Mechanisms involved in the association between hyperhomocysteinemia and vascular occlusive diseases remain unclear. Homocysteine (Hcy) may disturb calcium (Ca 2+ ) cytosolic regulation in endothelial cells, a process that can directly affect the synthesis of vasoactive substances, such as nitric oxide (NO). We have investigated the effect of acute and chronic incubation with high concentrations of Hcy (100 and 500 μmol/L) on the changes in the intracellular Ca 2+ concentration ([Ca 2+ ] i ) induced by ATP, using primary cultures of human umbilical vein endothelial cells (HUVEC). The changes in [Ca 2+ ] i , expressed as Δ F t / F b , were measured using the microspectrofluorimetric technique with Fluo‐3 as Ca 2+ indicator. HUVEC acutely exposed to Hcy did not produce significant effects on any of the parameters studied. However, chronic exposition (24 h) caused a significant decrease in the speed of store‐mediated Ca 2+ entry, expressed as (Δ F t / F b )/ t (s −1 ). Exposure of HUVEC to 100 and 500 µmol/L Hcy gave significantly lower values (0.019 ± 0.002 s −1 , n = 5 and 0.021 ± 0.004 s −1 , n = 6, respectively) compared to the controls (0.046 ± 0.004 s −1 , n = 8, P < 0.003). This was detected only when the sustained phase of the ATP‐induced [Ca +2 ] i increase was isolated. These results demonstrate that high concentrations of Hcy can affect the mechanisms involved in [Ca 2+ ] i regulation of HUVEC, and that alteration occurs specifically in the sustained phase, which has been directly associated with NO synthesis.