Protection against Nitric Oxide-Induced Apoptosis in Rat Mesangial Cells Demands Mitogen-Activated Protein Kinases and Reduced Glutathione
- University of Erlangen-Nürnberg, Faculty of Medicine, Department of Medicine IV, Experimental Division, Erlangen, Germany
Abstract
Inflammatory diseases such as proliferative glomerulonephritis are associated with the production of nitric oxide (NO), which can initiate apoptotic/necrotic cell death. We studied the role of the p42/44 mitogen-activated protein kinases (MAPKs) and c-Jun N-terminal kinases1/2 (JNK1/2) in NO-evoked cytotoxicity in rat mesangial cells (MC). The NO donor S-nitrosoglutathione time- and concentration-dependently promoted apoptotic cell death as detected by JNK1/2 and caspase-3 activation as well as DNA fragmentation. By using Ro 318220, a JNK1/2 activator, we established a correlation between apoptosis and JNK1/2 activation. Apoptosis is antagonized by the addition of fetal calf serum or the simultaneous generation of NO and superoxide (O2−), another biological inflammatory mediator. Fetal calf serum-induced protection required p42/44 MAPK activation as inhibition of the p42/44 MAPK pathway by the MAPK kinase-1 inhibitor PD 98059 attenuated MC protection. In contrast, cytoprotection by NO/O2− cogeneration demanded reduced glutathione but was p42/44 MAPK unrelated. Depletion of glutathione reversed NO/O2−-evoked survival to cell destruction and reinstalled JNK1/2 activity. In conclusion, different signal transduction pathways facilitate protection against NO-induced JNK1/2 activation and apoptosis in rat MC.
Footnotes
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Send reprint requests to: Dr. Bernhard Brüne, University of Erlangen-Nürnberg, Faculty of Medicine, Loschgestrasse 8, D-91054 Erlangen, Germany. E-mail:mfm423{at}rzmail.uni-erlangen.de
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↵1 These authors contributed equally to this work.
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This work was supported by the Deutsche Forschungsgemeinschaft (Br 999/8-1 and SFB 423, A5) and the European Community.
- Abbreviations:
- MC
- mesangial cells
- GSNO
- S-nitrosoglutathione
- DMNQ
- 2,3-dimethoxy-1,4-naphtoquinone
- IGF
- insulin growth factor
- DTT
- dithiothreitol
- NO
- nitric oxide
- O2−
- superoxide
- JNK1/2
- c-Jun N-terminal kinases1/2
- MAPK
- mitogen-activated protein kinases
- BSO
- l-buthionine-sulfoxamine
- LDH
- lactate dehydrogenase
- GSH
- glutathione
- MKP-1
- mitogen-activated protein kinase phosphatase-1
- GF
- growth factor
- MEK
- mitogen-activated protein kinase kinase
- DEVD
- N-Acetyl-Asp-Glu-Val-Asp-7
- AMC
- amino-4-methylcoumarin
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- Received June 7, 1999.
- Accepted July 14, 1999.
- The American Society for Pharmacology and Experimental Therapeutics
