Thrombin upregulates interleukin-8 in lung fibroblasts via cleavage of proteolytically activated receptor-I and protein kinase C-gamma activation.
Acute and chronic interstitial lung diseases are accompanied by evidence of inflammation and vascular injury. Thrombin activity in bronchoalveolar lavage fluid from such conditions is often increased, as well as interleukin (IL)-8. We observed that conditioned medium from lung fibroblasts exposed to thrombin has chemotactic activity for polymorphonuclear cells, and that this activity can be abolished by antibody to IL-8. We report that thrombin stimulates expression of IL-8 in human lung fibroblasts on both the messenger RNA and protein levels in a time- and dose-dependent manner. Stimulation of IL-8 expression by thrombin is inhibited by specific thrombin inhibitors. Synthetic thrombin receptor agonist peptide-14 mimics thrombin's stimulation of IL-8 expression in a dose-dependent manner consistent with the idea that upregulation of IL-8 by thrombin in human lung fibroblasts requires cleavage of proteolytically activated receptor-I. We demonstrate further that thrombin-induced IL-8 synthesis is regulated by protein kinase (PK) C. PKC-gamma may be involved in the upregulation of lung fibroblast IL-8 by thrombin because stimulation of lung fibroblasts with thrombin caused significant upregulation of PKC-gamma and because PKC-gamma antisense oligonucleotides inhibited the accumulation of PKC-gamma protein and IL-8 protein. Our data suggest that the PKC-gamma isoform increase observed after thrombin stimulation is required for thrombin-induced IL-8 formation by human lung fibroblasts.
Ludwicka-Bradley
A
A
Division of Rheumatology and Immunology, Department of Medicine, Medical University of South Carolina, Charleston, South Carolina 29425, USA.
Tourkina
E
E
Suzuki
S
S
Tyson
E
E
Bonner
M
M
Fenton
J W
JW
Hoffman
S
S
Silver
R M
RM
D001483:Base Sequence; D002633:Chemotaxis / Q000187:drug effects; D017077:Culture Media, Conditioned; D017931:DNA Primers; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D007527:Isoenzymes / Q000378:metabolism*; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D010446:Peptide Fragments / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018179:Receptors, Thrombin / Q000737:chemistry / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology / Q000502:physiology*; D015854:Up-Regulation
D017077:Culture Media, Conditioned; D017931:DNA Primers; D016209:Interleukin-8; D007527:Isoenzymes; D010446:Peptide Fragments; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D018179:Receptors, Thrombin; C072032:thrombin receptor peptide (42-55); C087857:protein kinase C gamma; D011493:Protein Kinase C; D013917:Thrombin