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  • Serine hydrolase inhibitors block necrotic cell death by preventing calcium overload of the mitochondria and permeability transition pore formation.

Serine hydrolase inhibitors block necrotic cell death by preventing calcium overload of the mitochondria and permeability transition pore formation.

The Journal of biological chemistry (2013-12-04)
Bogeon Yun, HeeJung Lee, Moumita Ghosh, Benjamin F Cravatt, Ku-Lung Hsu, Joseph V Bonventre, Heather Ewing, Michael H Gelb, Christina C Leslie
摘要

Perturbation of calcium signaling that occurs during cell injury and disease, promotes cell death. In mouse lung fibroblasts A23187 triggered mitochondrial permeability transition pore (MPTP) formation, lactate dehydrogenase (LDH) release, and necrotic cell death that were blocked by cyclosporin A (CsA) and EGTA. LDH release temporally correlated with arachidonic acid release but did not involve cytosolic phospholipase A2α (cPLA2α) or calcium-independent PLA2. Surprisingly, release of arachidonic acid and LDH from cPLA2α-deficient fibroblasts was inhibited by the cPLA2α inhibitor pyrrophenone, and another serine hydrolase inhibitor KT195, by preventing mitochondrial calcium uptake. Inhibitors of calcium/calmodulin-dependent protein kinase II, a mitochondrial Ca(2+) uniporter (MCU) regulator, also prevented MPTP formation and arachidonic acid release induced by A23187 and H2O2. Pyrrophenone blocked MCU-mediated mitochondrial calcium uptake in permeabilized fibroblasts but not in isolated mitochondria. Unlike pyrrophenone, the diacylglycerol analog 1-oleoyl-2-acetyl-sn-glycerol and CsA blocked cell death and arachidonic acid release not by preventing mitochondrial calcium uptake but by inhibiting MPTP formation. In fibroblasts stimulated with thapsigargin, which induces MPTP formation by a direct effect on mitochondria, LDH and arachidonic acid release were blocked by CsA and 1-oleoyl-2-acetyl-sn-glycerol but not by pyrrophenone or EGTA. Therefore serine hydrolase inhibitors prevent necrotic cell death by blocking mitochondrial calcium uptake but not the enzyme releasing fatty acids that occurs by a novel pathway during MPTP formation. This work reveals the potential for development of small molecule cell-permeable serine hydrolase inhibitors that block MCU-mediated mitochondrial calcium overload, MPTP formation, and necrotic cell death.

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Sigma-Aldrich
磷脂酶 A 2 from honey bee venom (Apis mellifera), salt-free, lyophilized powder, 600-2400 units/mg protein
Sigma-Aldrich
磷脂酶 A2 来源于猪胰腺, ammonium sulfate suspension, ≥600 units/mg protein
Sigma-Aldrich
磷脂酶 A 2 来源于牛胰腺, lyophilized powder, ≥20 units/mg protein
Avanti
18:1-2:0 DG, 1-oleoyl-2-acetyl-sn-glycerol, neat oil
Sigma-Aldrich
KT109, ≥98% (HPLC)
Avanti
18:1-2:0 DG, 1-oleoyl-2-acetyl-sn-glycerol, chloroform