Document Detail


Inhibition of nitric oxide and caspase-3 mediated apoptosis by a tetrapeptide derivative (PEP1261) in cultured synovial fibroblasts from collagen-induced arthritis.
MedLine Citation:
PMID:  16317520     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In this study, the effect of (Boc-Lys (Boc)-Arg-Asp-Ser (tBu)-OtBu), a tetrapeptide derivative (PEP1261) was examined for antiproliferative potency and apoptotic induction. Synovial fibroblasts were isolated from collagen-induced arthritic (CIA) rats and exposed to peptides viz., PEP1261, and parental peptides (KRDS and RGDS). Viability of the cells decreased in the presence of PEP1261 at a lower concentration (0.1 mM) when compared to RGDS and KRDS (1 mM). The treatment of cells with peptides showed induction of apoptosis, resulting in the cleavage of caspase-3 as well as its substrate poly-(ADP-ribose) polymerase (PARP). Pretreatment of cells with caspase-3 inhibitor prevented inhibition of [(3)H] thymidine incorporation, DNA fragmentation, and cleavage of caspase-3 and PARP as confirmed by western blotting as well as annexin-V/PI-staining using flow cytometry. However, caspase-1 and caspase-2 inhibitors did not prevent the peptides from inducing apoptosis indicating that caspase-3 might have a role in the process of apoptosis induced by peptides. Treatment of synovial fibroblasts with nitric oxide donor, S-nitroso-N-acetyl-DL: -penicillamine (SNAP) (500 microM) showed significant elevation of nitric oxide levels and resulted in absence of apoptosis by preventing the inhibition of [(3)H] thymidine incorporation. This was further evidenced by annexin V/propidium iodide (PI) staining and absence of DNA fragmentation, intra cellular caspase-3 activity and PARP cleavage. In contrast, SNAP followed by PEP1261 and parental peptides-induced apoptosis by lowering the levels of nitric oxide. These results suggested that PEP1261 suppressed the proliferation and induced apoptosis in cultured synovial fibroblasts from CIA rats. This study also confirmed that PEP1261 inhibited nitric oxide level in cultured synovial fibroblasts.
Authors:
Dilly Ashok Kumar; K Settu; Kalidindi V S Narayana Raju; K Kumanan; Bhakthavatchalam Murali Manohar; Rengarajulu Puvanakrishnan
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular and cellular biochemistry     Volume:  282     ISSN:  0300-8177     ISO Abbreviation:  Mol. Cell. Biochem.     Publication Date:  2006 Jan 
Date Detail:
Created Date:  2005-11-30     Completed Date:  2006-04-20     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0364456     Medline TA:  Mol Cell Biochem     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  125-39     Citation Subset:  IM    
Affiliation:
Department of Biotechnology, Central Leather Research Institute, Adyar, Chennai 600 020, India.
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MeSH Terms
Descriptor/Qualifier:
Animals
Annexin A5 / metabolism
Apoptosis / drug effects*
Arthritis / chemically induced,  metabolism*,  pathology
Caspase 3
Caspases / metabolism*
Cell Proliferation / drug effects
Cells, Cultured
Collagen Type II
Enzyme Activation
Fibroblasts / drug effects*,  metabolism,  pathology
Lactoferrin / pharmacology*
Male
Nitric Oxide / antagonists & inhibitors*,  metabolism
Oligopeptides / pharmacology
Peptide Fragments / pharmacology*
Poly(ADP-ribose) Polymerases / metabolism
Protein Binding
Rats
Rats, Wistar
Synovial Fluid / cytology
Chemical
Reg. No./Substance:
0/Annexin A5; 0/Boc-Lys(Boc)-Arg-Asp-Ser(tBu)-OtBu; 0/Collagen Type II; 0/Lactoferrin; 0/Oligopeptides; 0/Peptide Fragments; 10102-43-9/Nitric Oxide; 116430-80-9/KRDS peptide; 91037-65-9/arginyl-glycyl-aspartyl-serine; EC 2.4.2.30/Poly(ADP-ribose) Polymerases; EC 3.4.22.-/Casp3 protein, rat; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases

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