Fasciola hepatica proteolytic activity in liver revealed by in situ zymography.
Fasciola hepatica secretes cysteine proteases that play a role in facilitating parasite migration. The aim of this study was to detect the inhibition of the proteolytic activity of F. hepatica cysteine proteases in the liver of C57BL/6 cathepsin B knockout mice (cat B-/-) and wild-type controls (cat B+/+) by intraperitoneal administration of N-[N-(L-3-trans-carboxyoxirane-2-carbonyl)-L-leucyl]-agmatine, (E-64) using the film in situ zymography (FIZ) technique and image analysis. The FIZ technique revealed that intraperitoneal administration of E-64 dramatically reduced (85%) F. hepatica proteolytic activity in the liver of experimentally infected mice with no discernable side effects. These results suggest the usefulness of the FIZ for determining in vivo activity of F. hepatica proteases, as well as their inhibition by intraperitoneal administration of E-64 in hepatic tissue of infected mice.
Alcalá-Canto
Yazmín
Y
Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, México, DF 04510. yalcala@prodigy.net.mx
Ibarra-Velarde
Froylán
F
Gracia-Mora
Jesús
J
Sumano-López
Héctor
H
D000818:Animals; D002401:Cathepsin B / Q000172:deficiency* / Q000235:genetics; D002403:Cathepsins / Q000037:antagonists & inhibitors; D004796:Clinical Enzyme Tests / Q000379:methods; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D005210:Fasciola hepatica / Q000201:enzymology* / Q000502:physiology; D005211:Fascioliasis / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D007274:Injections, Intraperitoneal; D007930:Leucine / Q000008:administration & dosage / Q000031:analogs & derivatives; D008099:Liver / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout
D005780:Gelatin; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; D007930:Leucine; C024974:E 64