Role of calcium in pancreatic islet cell death by IFN-gamma/TNF-alpha.
We studied the intracellular events associated with pancreatic beta cell apoptosis by IFN-gamma/TNF-alpha synergism. IFN-gamma/TNF-alpha treatment of MIN6N8 insulinoma cells increased the amplitude of high voltage-activated Ca(2+) currents, while treatment with IFN-gamma or TNF-alpha alone did not. Cytosolic Ca(2+) concentration ([Ca(2+)](c)) was also increased by IFN-gamma/TNF-alpha treatment. Blockade of L-type Ca(2+) channel by nifedipine abrogated death of insulinoma cells by IFN-gamma/TNF-alpha. Diazoxide that attenuates voltage-activated Ca(2+) currents inhibited MIN6N8 cell death by IFN-gamma/TNF-alpha, while glibenclamide that accentuates voltage-activated Ca(2+) currents augmented insulinoma cell death. A protein kinase C inhibitor attenuated MIN6N8 cell death and the increase in [Ca(2+)](c) by IFN-gamma/TNF-alpha. Following the increase in [Ca(2+)](c), calpain was activated, and calpain inhibitors decreased insulinoma cell death by IFN-gamma/TNF-alpha. As a downstream of calpain, calcineurin was activated and the inhibition of calcineurin activation by FK506 diminished insulinoma cell death by IFN-gamma/TNF-alpha. BAD phosphorylation was decreased by IFN-gamma/TNF-alpha because of the increased calcineurin activity, which was reversed by FK506. IFN-gamma/TNF-alpha induced cytochrome c translocation from mitochondria to cytoplasm and activation of caspase-9. Effector caspases such as caspase-3 or -7 were also activated by IFN-gamma/TNF-alpha treatment. These results indicate that IFN-gamma/TNF-alpha synergism induces pancreatic beta cell apoptosis by Ca(2+) channel activation followed by downstream intracellular events such as mitochondrial events and caspase activation and also suggest the therapeutic potential of Ca(2+) modulation in type 1 diabetes.
Chang
Inik
I
Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
Cho
Namjoo
N
Kim
Sunshin
S
Kim
Ja Young
JY
Kim
Eunshil
E
Woo
Ji-Eun
JE
Nam
Joo Hyun
JH
Kim
Sung Joon
SJ
Lee
Myung-Shik
MS
D000818:Animals; D017209:Apoptosis; D019703:Calcineurin / Q000502:physiology; D002118:Calcium / Q000502:physiology*; D002352:Carrier Proteins / Q000378:metabolism; D020169:Caspases / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D007371:Interferon-gamma / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000473:pathology*; D051379:Mice; D016688:Mice, Inbred NOD; D010766:Phosphorylation; D021381:Protein Transport; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D051025:bcl-Associated Death Protein
C494820:Bad protein, mouse; D002352:Carrier Proteins; D014409:Tumor Necrosis Factor-alpha; D051025:bcl-Associated Death Protein; D007371:Interferon-gamma; D045304:Cytochromes c; D019703:Calcineurin; D020169:Caspases; D002118:Calcium