Pharmacological enhancement of the kallikrein-kinin system promotes anti-fibrotic responses in human mesangial cells.
The aim of the present study was to investigate whether pharmacological enhancement of the renal kallikrein-kinin system using the vasopeptidase inhibitor omapatrilat plays a direct role in modulating the fibrotic responses of human mesangial cells to injury. Treatment with 40 micromol/L omapatrilat was able to reduce macrophage-conditioned medium (MPCM)-induced fibronectin levels without affecting mRNA expression. MPCM injury also suppressed kallikrein and low molecular weight kininogen mRNA. Omapatrilat was able to attenuate this suppression. Bradykinin levels in contrast were increased by MPCM and treatment with omapatrilat further augmented levels. Co-incubation with the bradykinin B2 receptor antagonist HOE 140 attenuated the omapatrilat-induced lowering of fibronectin. Moreover, inhibition of cGMP release had a similar effect. Paradoxically, RT-PCR and Southern blotting demonstrated that bradykinin B2 receptor mRNA levels were down regulated in response to omapatrilat. Western blotting supported this data. Supernatant levels of tissue plasminogen activator (tPA), a product of bradykinin stimulation, were decreased by omapatrilat while cell associated tPA levels were increased. Matrix metalloproteinase-9 (MMP-9) mRNA expression was up regulated by omapatrilat treatment, although no difference in active zymogen levels was observed. In conclusion enhancement of kallikrein-kinin system appears to play a direct role in promoting anti-fibrotic responses in MPCM-injured human mesangial cells.
Pawluczyk
Izabella Z A
IZ
John Walls Renal Unit, Leicester General Hospital, UK. izapl@le.ac.uk
Patel
Samita R
SR
Harris
Kevin P G
KP
D001920:Bradykinin / Q000031:analogs & derivatives / Q000494:pharmacology; D065094:Bradykinin B2 Receptor Antagonists; D017077:Culture Media, Conditioned / Q000633:toxicity; D005353:Fibronectins / Q000378:metabolism; D005355:Fibrosis; D006801:Humans; D016234:Kallikrein-Kinin System / Q000187:drug effects* / Q000502:physiology; D007610:Kallikreins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D007704:Kininogens / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D050527:Mesangial Cells / Q000187:drug effects* / Q000473:pathology*; D011480:Protease Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D043783:Receptor, Bradykinin B1 / Q000235:genetics; D043782:Receptor, Bradykinin B2 / Q000235:genetics; D013841:Thiazepines / Q000494:pharmacology*; D010959:Tissue Plasminogen Activator / Q000032:analysis
D065094:Bradykinin B2 Receptor Antagonists; D017077:Culture Media, Conditioned; D005353:Fibronectins; D007704:Kininogens; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors; D011725:Pyridines; D012333:RNA, Messenger; D043783:Receptor, Bradykinin B1; D043782:Receptor, Bradykinin B2; D013841:Thiazepines; C106266:omapatrilat; C065679:icatibant; D007610:Kallikreins; D010959:Tissue Plasminogen Activator; D020780:Matrix Metalloproteinase 9; D001920:Bradykinin