Clostridium perfringens phospholipase C induced ROS production and cytotoxicity require PKC, MEK1 and NFκB activation.
Clostridium perfringens phospholipase C (CpPLC), also called α-toxin, is the most toxic extracellular enzyme produced by this bacteria and is essential for virulence in gas gangrene. At lytic concentrations, CpPLC causes membrane disruption, whereas at sublytic concentrations this toxin causes oxidative stress and activates the MEK/ERK pathway, which contributes to its cytotoxic and myotoxic effects. In the present work, the role of PKC, ERK 1/2 and NFκB signalling pathways in ROS generation induced by CpPLC and their contribution to CpPLC-induced cytotoxicity was evaluated. The results demonstrate that CpPLC induces ROS production through PKC, MEK/ERK and NFκB pathways, the latter being activated by the MEK/ERK signalling cascade. Inhibition of either of these signalling pathways prevents CpPLC's cytotoxic effect. In addition, it was demonstrated that NFκB inhibition leads to a significant reduction in the myotoxicity induced by intramuscular injection of CpPLC in mice. Understanding the role of these signalling pathways could lead towards developing rational therapeutic strategies aimed to reduce cell death during a clostridialmyonecrosis.
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Laura
L
Monturiol-Gross
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Marietta
M
Flores-Díaz
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Maria Jose
MJ
Pineda-Padilla
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Ana Cristina
AC
Castro-Castro
Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica ; Departamento de Bioquímica, Escuela de Medicina, Universidad de Costa Rica, San José, Costa Rica ; Centro de investigación en estructuras microscópicas, Universidad de Costa Rica, San José, Costa Rica.
Alberto
A
Alape-Giron
D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology*; D015153:Blotting, Western; D016466:CHO Cells; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D003412:Cricetulus; D048369:MAP Kinase Kinase 1 / Q000378:metabolism*; D008545:Melanoma / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology*; D016328:NF-kappa B / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000494:pharmacology*
D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D011493:Protein Kinase C; D048369:MAP Kinase Kinase 1; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens