Effect of plant neutrophil elastase inhibitor on leucocyte migration, adhesion and cytokine release in inflammatory conditions.
OBJECTIVE
The serine and cysteine peptidase inhibitor, BbCI, isolated from Bauhinia bauhinioides seeds, is similar to the classical plant Kunitz inhibitor, STI, but lacks disulphide bridges and methionine residues. BbCI blocks activity of the serine peptidases, elastase (K(iapp) 5.3 nM) and cathepsin G (K(iapp) 160.0 nM), and the cysteine peptidase cathepsin L (K(iapp) 0.2 nM). These three peptidases play important roles in the inflammatory process.


METHODS
We measured the effects of BbCI on paw oedema and on leucocyte accumulation in pleurisy, both induced by carrageenan. Leucocyte-endothelial cell interactions in scrotal microvasculature in Wistar rats were investigated using intravital microscopy. Cytokine levels in pleural exudate and serum were measured by elisa.


RESULTS
Pretreatment of the animals with BbCI (2.5 mg·kg(-1)), 30 min before carrageenan-induced inflammation, effectively reduced paw oedema and bradykinin release, neutrophil migration into the pleural cavity. The number of rolling, adhered and migrated leucocytes at the spermatic fascia microcirculation following carrageenan injection into the scrotum were reduced by BbCI pretreatment. Furthermore, levels of the rat chemokine cytokine-induced neutrophil chemo-attractant-1 were significantly reduced in both pleural exudates and serum from animals pretreated with BbCI. Levels of interleukin-1β or tumour necrosis factor-α, however, did not change.


CONCLUSIONS
Taken together, our data suggest that the anti-inflammatory properties of BbCI may be useful in investigations of other pathological processes in which human neutrophil elastase, cathepsin G and cathepsin L play important roles.
Oliveira
C
C
Department of Biochemistry, Universidade Federal de São Paulo, Rua Três de Maio, São Paulo, Brazil.
Navarro-Xavier
R A
RA
Anjos-Vallota
E A
EA
Martins
J O
JO
Silveira
V L F
VL
Gonçalves
L R C
LR
Araújo
M S
MS
Motta
G
G
Sannomiya
P
P
Oliva
M L V
ML
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000302:isolation & purification / Q000494:pharmacology*; D036481:Bauhinia / Q000737:chemistry; D002351:Carrageenan; D056649:Cathepsin G / Q000037:antagonists & inhibitors / Q000378:metabolism; D056668:Cathepsin L / Q000037:antagonists & inhibitors / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004487:Edema / Q000188:drug therapy / Q000503:physiopathology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007249:Inflammation / Q000188:drug therapy* / Q000503:physiopathology; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors / Q000378:metabolism; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008853:Microscopy / Q000379:methods; D010940:Plant Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D012639:Seeds
D000893:Anti-Inflammatory Agents; C505411:BbCI protein, Bauhinia bauhinoides; D016207:Cytokines; D010940:Plant Proteins; D002351:Carrageenan; D056649:Cathepsin G; D019272:Leukocyte Elastase; D056668:Cathepsin L