Up regulation of interleukin-8 expressions induced by mast cell tryptase via protease activated receptor-2 in endothelial cell line.
BACKGROUND
Protease activated receptor-2 is cleaved and activated by trypsin or mast cell tryptase and may play an important role in inflammation. However, it is unknown whether PAR-2 can mediate tryptase-induced inflammatory reaction. This study was conduct to investigate whether PAR-2 could be the activated by mast cell tryptase and medicated the tryptase induced interleukin-8 expression in endothelial cells.


METHODS
Protease activated receptor-2 expression was found in endothelial cell lines ECV304 cell by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Interleukin-8 stimulated by purified human mast cell tryptase was determined by RT-PCR and enzyme linked immunosorbent assay (ELISA). Data were analysed by the S-N-K one-way ANOVA test.


RESULTS
The present study shows that mRNA and protein of protease activated receptor-2 could be expressed in ECV304 cells, and tryptase upregulated the expression levels of both interleukin-8 mRNA and protein. The increased expression of interleukin-8 was inhibited by an antiprotease activated receptor-2 monoclonal antibody, SAM11. An additional band was observed by Western blotting after the incubation of ECV304 cells with tryptase for 2 hours, which suggested that protease activated receptor-2 was activated.


CONCLUSIONS
Protease activated receptor-2 can mediate the mast cell tryptase stimulated expression of interleukin-8 in ECV304 cell.
Lu
Chao
C
Department of Physiology and Pathophysiology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
Zhao
Feng-di
FD
Li
Xiao-Bo
XB
Yin
Lian-Hua
LH
D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D044464:Receptor, PAR-2 / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D012697:Serine Endopeptidases / Q000502:physiology*; D053802:Tryptases; D015854:Up-Regulation
D000911:Antibodies, Monoclonal; D016209:Interleukin-8; D012333:RNA, Messenger; D044464:Receptor, PAR-2; D012697:Serine Endopeptidases; D053802:Tryptases