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Acta Physiologica 2008; Volume 194, Supplement 665
The 59th National Congress of the Italian Physiological Society
9/17/2008-9/19/2008
Cagliari, Italy
EFFECT OF ANG II ON [CA2+]I LEVELS IN HUVEC CULTURES
Abstract number: P88
MARTINI1 A, MAZZULLA1 S, BRUNO2 R, NOCITA1 A, MARTINO1 G
1Dept.of Cell Biology, Calabria University - Via P. Bucci 4C, 87030 Rende (CS), Italy
2Dept. of Pharmaco Biology, Calabria University - Via P. Bucci, 87030 Rende (CS), [email protected]
Aim:
Angiotensin II (Ang II) is involved in vascular smooth muscle tone.The aim is to investigate the effects of Ang II on calcium levels of HUVECs
(Human umbelical vein endothelial cells) and by what kind of pathways.
Methods:
HUVEC were obtained in their passages 3–8, grown in EGM2 Bullet Kit (LONZA) containing 10% FBS and grow factors. The confluent cells were trypsinized and treated with Ang II at 10-7 M with or without Wortmannin (Wort), inhibitor of PI3-K/Akt pathway or Efonidipine, T-Type Calcium channel blocker. All chemicals were from Sigma Aldrich.The cells are incubated with Fluo-3AM (Invitrogen-Molecular Probes), fluorescent for the determination of intracellular Ca2+ by microfluorometry (20X). The data evaluation is at 3, 6 and 9 hours. The images were captured by ProImagePlus on Olympus LX 50 microscope and their analysis was performed by NIH imageJ program.
Results:
The IP3 pathway is significantly involved in [Ca2+]i release, infact Wort 10-7 M decreases HUVECs calcium levels in comparison to Ang II. The use of Efonidipine shows that also plasma membrane calcium channels are involved, infact even more significantly at 3 h, and 6 h, and 9 h.
Conclusions:
The Ang II acts by both IP3 pathways and by plasma membrane T-type Calcium channel.
To cite this abstract, please use the following information:
Acta Physiologica 2008; Volume 194, Supplement 665 :P88