Document Detail


Kallikrein gene 'knock-down' by small interfering RNA transfection induces a profibrotic phenotype in rat mesangial cells.
MedLine Citation:
PMID:  18090545     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Emerging evidence suggests that kallikrein exerts renoprotective effects independent of its haemodynamic actions. The aim of the current investigation was to delineate the role of kallikrein in the regulation of fibrosis, by 'knocking down' its expression using specific small interfering RNAs (siRNA). METHODS: Rat mesangial cells were treated with 12, 60, 120 nmol/l kallikrein-specific siRNAs. The consequent cellular genotypes and phenotypes were analysed. RESULTS: Western blotting demonstrated that mesangial cells produced a kallikrein protein, which was of a different molecular weight to urinary kallikrein from rats of the same species. Treatment of cells with siRNA resulted in a dose-dependent decrease in kallikrein mRNA levels, which impacted on other components of the kallikrein-kinin system, dose-dependently reducing bradykinin B2 receptor mRNA expression. Kallikrein suppression resulted in significant increases in fibronectin and transforming growth factor-beta protein levels in culture supernatants over control levels. Gelatin zymography demonstrated a siRNA dose-dependent decrease in active MMP-2 enzyme levels. Bradykinin, an effector molecule of the kallikrein system, is known to stimulate tissue plasminogen activator production. Paradoxically, however, tissue plasminogen activator protein levels were augmented with increasing kallikrein mRNA silencing. This was accompanied by a dose-dependent decrease in low-density lipoprotein receptor-related protein mRNA levels, indicating that increased tissue plasminogen activator levels were due to an attenuation of receptor-mediated protease clearance. CONCLUSION: These data lend strong support to the hypothesis that kallikrein exerts antifibrotic, renoprotective effects that are independent of its classical haemodynamic actions.
Authors:
Izabella Z A Pawluczyk; Eddie K C Tan; David Lodwick; Kevin P G Harris
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of hypertension     Volume:  26     ISSN:  0263-6352     ISO Abbreviation:  J. Hypertens.     Publication Date:  2008 Jan 
Date Detail:
Created Date:  2007-12-19     Completed Date:  2008-08-28     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8306882     Medline TA:  J Hypertens     Country:  England    
Other Details:
Languages:  eng     Pagination:  93-101     Citation Subset:  IM    
Affiliation:
Department of Infection, Immunity and Inflammation, University of Leicester, UK. izap1@le.ac.uk
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
Cells, Cultured
Culture Media, Conditioned / pharmacology
Dose-Response Relationship, Drug
Female
Fibronectins / drug effects,  metabolism
Fibrosis / chemically induced,  genetics*
Gene Expression Regulation, Enzymologic / drug effects,  genetics
LDL-Receptor Related Protein 1 / drug effects,  genetics,  metabolism
Male
Matrix Metalloproteinase 2 / drug effects,  metabolism
Mesangial Cells / drug effects*,  metabolism
Phenotype
RNA, Messenger / antagonists & inhibitors,  genetics
RNA, Small Interfering / pharmacology*
Rats
Rats, Wistar
Receptor, Bradykinin B2 / drug effects,  genetics
Tissue Kallikreins / biosynthesis,  drug effects*,  genetics*
Transfection
Transforming Growth Factor beta / drug effects,  metabolism
Chemical
Reg. No./Substance:
0/Culture Media, Conditioned; 0/Fibronectins; 0/LDL-Receptor Related Protein 1; 0/RNA, Messenger; 0/RNA, Small Interfering; 0/Receptor, Bradykinin B2; 0/Transforming Growth Factor beta; EC 3.4.21.35/Tissue Kallikreins; EC 3.4.24.24/Matrix Metalloproteinase 2

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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