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<b><font color="#3366FF"><font size=+4>Journal of APPLIED BIOMEDICINE</font></font></b><br>
ISSN 1214-0287 (on-line)<br>
ISSN 1214-021X (printed)
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Volume 6 (2008), No 4, p 211-224
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Role of intracellular calcium on hydrogen peroxide-induced apoptosis in rat pancreatic acinar AR42J cells
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Sara Morgado, Maria P. Granados, Ignacio Bejarano, Jose J. Lopez, Gines M. Salido, Antonio Gonzalez, Jose A. Pariente
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Address: J. A. Pariente, Department of Physiology, University of Extremadura, 06071 Badajoz, Spain
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<a href="mailto:pariente@unex.es">pariente@unex.es </a>
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Received 23rd October 2008.<br>
Revised 12th November 2008.<br>
Published online 12th December 2008.<br>
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<a name="summary"><b>SUMMARY</b></a><br>
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.
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<a name="keywords"><b>KEY WORDS</b></a><br>
programed cell death; casase-3; cytochrome c;mitochondrion; thapsingargin; dimethyl-BAPTA; Ru360;
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