PKD prevents H2O2-induced apoptosis via NF-kappaB and p38 MAPK in RIE-1 cells.

Previously, we demonstrated that protein kinase D (PKD) plays a protective role during H(2)O(2)-induced intestinal cell death. Here, we sought to determine whether this effect is mediated by nuclear factor-kappaB (NF-kappaB) and mitogen-activated protein kinases (MAPKs). Treatment with H(2)O(2) activated NF-kappaB in RIE-1 cells; H(2)O(2) also induced the translocation of NF-kappaB p65 as well as phosphorylation of IkappaB-alpha. PKD1 siRNA inhibited H(2)O(2)-induced activation, translocation of NF-kappaB, and phosphorylation of IkappaB-alpha. We also found that overexpression of wild type PKD1 attenuated H(2)O(2)-induced phosphorylation of p38 MAPK and its upstream activator, MAPK kinase (MKK) 3/6, whereas the phosphorylation was increased by PKD1 siRNA or kinase-dead PKD1. Phosphorylation of neither extracellular signal-regulated kinases (ERK) 1/2 nor c-Jun N-terminal kinases (JNK) was altered by PKD1 plasmids or siRNA. Our findings suggest that PKD protects intestinal cells through up-regulation of NF-kappaB and down-regulation of p38 MAPK.

[1]  D. Chung,et al.  Molecular Mechanisms Contributing to Necrotizing Enterocolitis , 2001, Annals of surgery.

[2]  D. Chung,et al.  Oxidative stress-induced intestinal epithelial cell apoptosis is mediated by p38 MAPK. , 2006, Biochemical and biophysical research communications.

[3]  Michael Karin,et al.  Is NF‐κB the sensor of oxidative stress? , 1999 .

[4]  Peter Storz,et al.  Protein Kinase D Mediates Mitochondrion-to-Nucleus Signaling and Detoxification from Mitochondrial Reactive Oxygen Species , 2005, Molecular and Cellular Biology.

[5]  Roger J. Davis,et al.  Selective Activation of p38 Mitogen-activated Protein (MAP) Kinase Isoforms by the MAP Kinase Kinases MKK3 and MKK6* , 1998, The Journal of Biological Chemistry.

[6]  J. Avruch,et al.  Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. , 2001, Physiological reviews.

[7]  S. Harrison,et al.  Structure of an IκBα/NF-κB Complex , 1998, Cell.

[8]  Wei Zhang,et al.  Protein Kinase D Specifically Mediates Apoptosis Signal-regulating Kinase 1-JNK Signaling Induced by H2O2 but Not Tumor Necrosis Factor* , 2005, Journal of Biological Chemistry.

[9]  J. Blay,et al.  Characterization of an epithelioid cell line derived from rat small intestine: demonstration of cytokeratin filaments. , 1984, Cell biology international reports.

[10]  D. Chung,et al.  Signal Transduction Pathways Involved in Oxidative Stress-Induced Intestinal Epithelial Cell Apoptosis , 2005, Pediatric Research.

[11]  Peter Storz,et al.  Protein kinase D mediates a stress‐induced NF‐κB activation and survival pathway , 2003, The EMBO journal.

[12]  P. Baeuerle,et al.  Function and activation of NF-kappa B in the immune system. , 1994, Annual review of immunology.

[13]  M. Mattson,et al.  Activation of NF‐κB protects hippocampal neurons against oxidative stress‐induced apoptosis: Evidence for induction of manganese superoxide dismutase and suppression of peroxynitrite production and protein tyrosine nitration , 1997, Journal of neuroscience research.

[14]  C. Culmsee,et al.  Roles of Nuclear Factor κB in Neuronal Survival and Plasticity , 2000 .

[15]  T. Iglesias,et al.  Phosphorylation‐dependent protein kinase Dactivation , 1999, Electrophoresis.

[16]  K. Hensley,et al.  Redox regulatory mechanisms of cellular signal transduction. , 2000, Archives of biochemistry and biophysics.

[17]  A. Toker,et al.  Protein Kinase C Selectively Regulates Protein Kinase D-Dependent Activation of NF- B in Oxidative Stress Signaling , 2004 .

[18]  Á. Valverde,et al.  Molecular cloning and characterization of protein kinase D: a target for diacylglycerol and phorbol esters with a distinctive catalytic domain. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[19]  M. Karin,et al.  Mammalian MAP kinase signalling cascades , 2001, Nature.

[20]  N. Holbrook,et al.  Cellular response to oxidative stress: Signaling for suicide and survival * , 2002, Journal of cellular physiology.

[21]  D. Chung,et al.  Protein kinase D protects against oxidative stress-induced intestinal epithelial cell injury via Rho/ROK/PKC-δ pathway activation , 2006 .