Mitochondrial dysfunction links ceramide activated HRK expression and cell death.

<h4>Purpose</h4>Cell death is an essential process in normal development and homeostasis. In eyes, corneal epithelial injury leads to the death of cells in underlying stroma, an event believed to initiate corneal wound healing. The molecular basis of wound induced corneal stromal cell de...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Farhan Rizvi, Tom Heimann, Anja Herrnreiter, William J O'Brien
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/4dce8e16114846b1b0660ceb7b90ec18
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:4dce8e16114846b1b0660ceb7b90ec18
record_format	dspace
spelling	oai:doaj.org-article:4dce8e16114846b1b0660ceb7b90ec182021-11-18T06:56:22ZMitochondrial dysfunction links ceramide activated HRK expression and cell death.1932-620310.1371/journal.pone.0018137https://doaj.org/article/4dce8e16114846b1b0660ceb7b90ec182011-03-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21483866/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Purpose</h4>Cell death is an essential process in normal development and homeostasis. In eyes, corneal epithelial injury leads to the death of cells in underlying stroma, an event believed to initiate corneal wound healing. The molecular basis of wound induced corneal stromal cell death is not understood in detail. Studies of others have indicated that ceramide may play significant role in stromal cell death following LASIK surgery. We have undertaken the present study to investigate the mechanism of death induced by C6 ceramide in cultures of human corneal stromal (HCSF) fibroblasts.<h4>Methods</h4>Cultures of HCSF were established from freshly excised corneas. Cell death was induced in low passage (p<4) cultures of HCSF by treating the cells with C6 ceramide or C6 dihydroceramide as a control. Cell death was assessed by Live/Dead cell staining with calcein AM and ethidium homodimer-1 as well as Annexin V staining, caspase activation and TUNEL staining Mitochondrial dysfunction was assessed by Mito Sox Red, JC-1 and cytochrome C release Gene expression was examined by qPCR and western blotting.<h4>Results</h4>Our data demonstrate ceramide caused mitochondrial dysfunction as evident from reduced MTT staining, cyto c release from mitochondria, enhanced generation of ROS, and loss in mitochondrial membrane potential (ΔΨm). Cell death was evident from Live -Dead Cell staining and the inability to reestablish cultures from detached cells. Ceramide induced the expression of the harikari gene(HRK) and up-regulated JNK phosphorylation. In ceramide treated cells HRK was translocated to mitochondria, where it was found to interact with mitochondrial protein p32. The data also demonstrated HRK, p32 and BAD interaction. Ceramide-induced expression of HRK, mitochondrial dysfunction and cell death were reduced by HRK knockdown with HRK siRNA.<h4>Conclusion</h4>Our data document that ceramide is capable of inducing death of corneal stromal fibroblasts through the induction of HRK mediated mitochondria dysfunction.Farhan RizviTom HeimannAnja HerrnreiterWilliam J O'BrienPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 3, p e18137 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Farhan Rizvi Tom Heimann Anja Herrnreiter William J O'Brien Mitochondrial dysfunction links ceramide activated HRK expression and cell death.
description	<h4>Purpose</h4>Cell death is an essential process in normal development and homeostasis. In eyes, corneal epithelial injury leads to the death of cells in underlying stroma, an event believed to initiate corneal wound healing. The molecular basis of wound induced corneal stromal cell death is not understood in detail. Studies of others have indicated that ceramide may play significant role in stromal cell death following LASIK surgery. We have undertaken the present study to investigate the mechanism of death induced by C6 ceramide in cultures of human corneal stromal (HCSF) fibroblasts.<h4>Methods</h4>Cultures of HCSF were established from freshly excised corneas. Cell death was induced in low passage (p<4) cultures of HCSF by treating the cells with C6 ceramide or C6 dihydroceramide as a control. Cell death was assessed by Live/Dead cell staining with calcein AM and ethidium homodimer-1 as well as Annexin V staining, caspase activation and TUNEL staining Mitochondrial dysfunction was assessed by Mito Sox Red, JC-1 and cytochrome C release Gene expression was examined by qPCR and western blotting.<h4>Results</h4>Our data demonstrate ceramide caused mitochondrial dysfunction as evident from reduced MTT staining, cyto c release from mitochondria, enhanced generation of ROS, and loss in mitochondrial membrane potential (ΔΨm). Cell death was evident from Live -Dead Cell staining and the inability to reestablish cultures from detached cells. Ceramide induced the expression of the harikari gene(HRK) and up-regulated JNK phosphorylation. In ceramide treated cells HRK was translocated to mitochondria, where it was found to interact with mitochondrial protein p32. The data also demonstrated HRK, p32 and BAD interaction. Ceramide-induced expression of HRK, mitochondrial dysfunction and cell death were reduced by HRK knockdown with HRK siRNA.<h4>Conclusion</h4>Our data document that ceramide is capable of inducing death of corneal stromal fibroblasts through the induction of HRK mediated mitochondria dysfunction.
format	article
author	Farhan Rizvi Tom Heimann Anja Herrnreiter William J O'Brien
author_facet	Farhan Rizvi Tom Heimann Anja Herrnreiter William J O'Brien
author_sort	Farhan Rizvi
title	Mitochondrial dysfunction links ceramide activated HRK expression and cell death.
title_short	Mitochondrial dysfunction links ceramide activated HRK expression and cell death.
title_full	Mitochondrial dysfunction links ceramide activated HRK expression and cell death.
title_fullStr	Mitochondrial dysfunction links ceramide activated HRK expression and cell death.
title_full_unstemmed	Mitochondrial dysfunction links ceramide activated HRK expression and cell death.
title_sort	mitochondrial dysfunction links ceramide activated hrk expression and cell death.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/4dce8e16114846b1b0660ceb7b90ec18
work_keys_str_mv	AT farhanrizvi mitochondrialdysfunctionlinksceramideactivatedhrkexpressionandcelldeath AT tomheimann mitochondrialdysfunctionlinksceramideactivatedhrkexpressionandcelldeath AT anjaherrnreiter mitochondrialdysfunctionlinksceramideactivatedhrkexpressionandcelldeath AT williamjobrien mitochondrialdysfunctionlinksceramideactivatedhrkexpressionandcelldeath
_version_	1718424165328879616

Mitochondrial dysfunction links ceramide activated HRK expression and cell death.

Ejemplares similares