TABLE 3.
Effect of protease inhibitors on DPPIV activity on IL-1-stimulated fibroblastsa
| Inhibitor | Concn | % Inhibitionb |
|---|---|---|
| Diprotin A | 1 mM | 88.2 |
| PMSF | 1 mM | 42.9 |
| 0.5 mM | 26.9 | |
| Bestatin | 1 mM | 5.1 |
| Aprotinin | 10 μg/ml | <1 |
| Pepstatin | 10 μg/ml | <1 |
| Leupeptin | 20 μg/ml | <1 |
| 1,10-Phenanthroline | 1 mM | <1 |
| EDTA | 5 mM | 7.6 |
Confluent HGF (donor, FE) were stimulated with 10 ng of IL-1α per ml for 6 days in 96-well plates and then washed with PBS three times, and confluent-monolayer cells were examined by DPPIV assay in the presence or absence of the indicated inhibitors.
Percent inhibition was calculated as [(DPPIV activity on IL-1-stimulated HGF in the absence of inhibitor) − (DPPIV activity on IL-1-stimulated HGF in the presence of inhibitor)]/(DPPIV activity on IL-1-stimulated HGF in the absence of inhibitor) × 100. DPPIV activities of fibroblasts stimulated with or without IL-1α were 33.4 and 9.3 nmol/60 min/culture, respectively. The results are representative of three different experiments with three different donors (FE, NIK, and MH-12).