PMID- 15780084
OWN - NLM
STAT- MEDLINE
DCOM- 20050926
LR  - 20061115
IS  - 0085-2538 (Print)
IS  - 0085-2538 (Linking)
VI  - 67
IP  - 4
DP  - 2005 Apr
TI  - Thrombin stimulates proinflammatory and proliferative responses in primary 
      cultures of human proximal tubule cells.
PG  - 1315-29
AB  - BACKGROUND: Fibrin deposition is frequently observed within the 
      tubulointerstitium in various forms of chronic renal disease. This suggests the 
      presence of active components of the coagulation pathway, which may contribute to 
      the progressive deterioration in renal function. The aim of this study was to 
      investigate the proinflammatory and fibroproliferative effects of the coagulation 
      protease thrombin on human proximal tubular cells (PTC) in culture. METHODS: 
      Primary cultures of PTC were established from normal kidney tissue and grown 
      under serum-free conditions with or without thrombin or the protease-activated 
      receptor (PAR) activating peptides TFLLRN-NH(2), SLIGKV-NH(2), and SFLLRN-NH(2) 
      (100 to 400 micromol/L). DNA synthesis (thymidine incorporation), intracellular 
      Ca(2+) mobilization (fura-2 fluorimetry), fibronectin secretion [enzyme-linked 
      immunosorbent assay (ELISA), immunoblotting], monocyte chemoattractant protein-1 
      (MCP-1) secretion (ELISA), and transforming growth factor-beta1 (TGF-beta1) 
      secretion (ELISA) were measured. Reverse transcription-polymerase chain reaction 
      (RT-PCR) was used to assess PAR mRNA expression in these cells. RESULTS: Thrombin 
      enhanced DNA synthesis, fibronectin secretion, MCP-1 secretion, and TGF-beta1 
      secretion in a concentration-dependent manner. Cell injury [lactate dehydrogenase 
      (LDH) release] and cellular protein levels were unaffected. RT-PCR showed that 
      cultures of PTC expressed mRNA transcripts for the thrombin receptors PAR-1 and 
      PAR-3, but not PAR-4. Thrombin and each of the PAR activating peptides enhanced 
      intracellular calcium mobilization. However, the other effects of thrombin were 
      only fully reproduced by the PAR-2-specific peptide, SLIGKV-NH(2), only partially 
      by SFLLRN-NH(2), (a PAR-1 peptide that can activate PAR-2), and not at all by the 
      PAR-1-specific peptide, TFLLRN-NH(2). Thrombin-induced DNA synthesis, 
      fibronectin, and MCP-1 secretion were unaffected by a TGF-beta neutralizing 
      antibody, the matrix metalloproteinase (MMP) inhibitor, GM6001 and the epidermal 
      growth factor (EGF) receptor kinase inhibitor AG1478. CONCLUSION: Thrombin 
      initiates both proinflammatory and fibroproliferative responses in human PTC. 
      These responses which are dependent on its protease activity appear not to be 
      mediated by PAR-1 activation, the autocrine action of thrombin-induced TGF-beta1 
      secretion, MMP activation, or EGF receptor transactivation. The proinflammatory 
      and fibroproliferative actions of thrombin on human PTC may help explain the 
      extent of tubulointerstitial fibrosis observed in kidney diseases where fibrin 
      deposition is evident.
FAU - Vesey, David A
AU  - Vesey DA
AD  - Department of Renal Medicine, University of Queensland, Princess Alexandra 
      Hospital, Brisbane, Queenlsand, Australia. david_vesey@health.qld.gov.au
FAU - Cheung, Catherine W
AU  - Cheung CW
FAU - Kruger, Wade A
AU  - Kruger WA
FAU - Poronnik, Philip
AU  - Poronnik P
FAU - Gobe, Glenda
AU  - Gobe G
FAU - Johnson, David W
AU  - Johnson DW
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Kidney Int
JT  - Kidney international
JID - 0323470
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Oligopeptides)
RN  - 0 (Receptors, Proteinase-Activated)
RN  - 9007-49-2 (DNA)
RN  - EC 3.4.21.5 (Thrombin)
SB  - IM
MH  - Calcium Signaling/drug effects/physiology
MH  - Cell Division/drug effects
MH  - Cells, Cultured
MH  - Culture Media, Serum-Free
MH  - DNA/biosynthesis/drug effects
MH  - Humans
MH  - Inflammation
MH  - Kidney Tubules, Proximal/*cytology/drug effects/physiology
MH  - Oligopeptides/pharmacology
MH  - Receptors, Proteinase-Activated/drug effects/physiology
MH  - Thrombin/*pharmacology
EDAT- 2005/03/23 09:00
MHDA- 2005/09/27 09:00
CRDT- 2005/03/23 09:00
PHST- 2005/03/23 09:00 [pubmed]
PHST- 2005/09/27 09:00 [medline]
PHST- 2005/03/23 09:00 [entrez]
AID - S0085-2538(15)50587-5 [pii]
AID - 10.1111/j.1523-1755.2005.00209.x [doi]
PST - ppublish
SO  - Kidney Int. 2005 Apr;67(4):1315-29. doi: 10.1111/j.1523-1755.2005.00209.x.