ISSN 1214-0287 (on-line)
ISSN 1214-021X (printed)

Volume 6 (2008), No 4, p 211-224

Role of intracellular calcium on hydrogen peroxide-induced apoptosis in rat pancreatic acinar AR42J cells

Sara Morgado, Maria P. Granados, Ignacio Bejarano, Jose J. Lopez, Gines M. Salido, Antonio Gonzalez, Jose A. Pariente

Address: J. A. Pariente, Department of Physiology, University of Extremadura, 06071 Badajoz, Spain

Received 23rd October 2008.
Revised 12th November 2008.
Published online 12th December 2008.

Full text article (pdf)

The authors investigated whether cytosolic free calcium concentration ([Ca2+]c) plays a role in hydrogen peroxide-induced pancreatic acinar AR42J cells apoptosis. We analysed mitochondrial depolarization, [Ca2+]c determination and caspase-3 activity by fluorimetric methods, and cytochrome c release by subcellular fractionation and western blotting. The data shown that hydrogen peroxide, which causes a sustained [Ca2+]c increase, induces mitochondrial depolarization and cytochrome c release, and activation of caspase-3. Dimethyl-BAPTA loading did not affect hydrogen peroxide-evoked mitochondrial apoptosis, suggesting that these responses are independent of increases in [Ca2+]c. Treatment with thapsigargin, to induce extensive calcium store depletion and subsequent increases in [Ca2+]c, also stimulates mitochondrial depolarization cytochrome c release, and caspase-3 activation. Similar results were observed in AR42J cells loaded with dimethyl-BAPTA, suggesting that activation of apoptosis by thapsigargin does not require rises in [Ca2+]c. However, the blockade of mitochondrial calcium entry by pretreating with Ru360 showed protection against hydrogen peroxide- and thapsigargin-induced mitochondrial apoptosis. These results indicate that the apoptosis evoked y hydrogen peroxide and thapsigargin is mediated by mitochondrial calcium uptake.

programed cell death; caspase-3; cytochrome c; mitochondrion; thapsingargin; dimethyl-BAPTA; Ru360

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