Inhibition of cathepsin B does not affect the intracellular activation of trypsinogen by cerulein hyperstimulation in isolated rat pancreatic acinar cells.
Activation of trypsinogen is thought to trigger the autodigestive process in acute pancreatitis. The lysosomal enzyme cathepsin B was suggested to cause the activation of trypsinogen because it is known that cathepsin B is able to activate trypsinogen in special circumstances and that lysosomal and digestive enzymes are colocalized within intracellular vacuoles in the early stage of pancreatitis. As yet this hypothesis has been difficult to prove because activated trypsin is difficult to quantify in pancreatitis by conventional enzymatic measurements. We therefore employed an ELISA for trypsin activating peptide (TAP), which is a small peptide cleaved during the activation of trypsinogen and can be determined reliably. Supraphysiological concentrations of cerulein (1 nM-1 microM) resulted in a marked increase in TAP in freshly isolated pancreatic acinar cells, indicating activation of trypsinogen. This activation as determined by the TAP increase was significantly reduced by the serine protease inhibitor Fut-175 but not by the cathepsin B inhibitors E-64 and NCO-700. The concentrations of NCO-700 and E-64 abolished the cathepsin B activity of pancreatic acinar cells but did not significantly reduce the trypsin activity (after enterokinase preincubation); correspondingly the concentrations of Fut-175 used abolished the trypsin activity but did not reduce the cathepsin B activity. The results indicate that an autoactivation of trypsin rather than an activation of trypsinogen by cathepsin B triggers trypsin activation by supramaximal cerulein concentrations.
Klonowski-Stumpe
H
H
Department of Medicine, Heinrich-Heine-University of Düsseldorf, Germany.
Lüthen
R
R
Han
B
B
Sata
N
N
Häussinger
D
D
Niederau
C
C
D000818:Animals; D001550:Benzamidines; D002401:Cathepsin B / Q000037:antagonists & inhibitors*; D002108:Ceruletide / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005765:Gastrointestinal Agents / Q000494:pharmacology*; D006146:Guanidines / Q000494:pharmacology; D008297:Male; D010179:Pancreas / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D017208:Rats, Wistar; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism
D001550:Benzamidines; D005765:Gastrointestinal Agents; D006146:Guanidines; D015842:Serine Proteinase Inhibitors; D002108:Ceruletide; D014357:Trypsin; D002401:Cathepsin B; C032855:nafamostat