Table 3.
Effects of DPI analogs on superoxide and H2O2 production in human cells. PMA-induced superoxide production was measured by luminescence assay in HEK293 NOX1, PMNs (NOX2), HL-60 (NOX2), and UACC-257 (NOX5) cells. Hydrogen peroxide production was measured by Amplex Red assay of DPI or analog-treated NOX4 and DUOX2/DUOXA2 overexpression cell lines. The cells were incubated with DPI analogs for 30 min prior to assay measurement. The data represent the mean ± standard deviation (SD) of at least 3 independent experiments.
| Superoxide (O2•−) | Hydrogen peroxide (H2O2) | |||||
|---|---|---|---|---|---|---|
|
| ||||||
| HEK293 NOX1 | PMN (NOX2) | HL-60 (NOX2) | UACC-257 (NOX5) | HEK293 NOX4 | HEK293 DUOX2/DUOX A2 | |
| Baseline ROS producti on rate | 9.88 × 10−2 RLU/min/c ell | 0.55 RLU/min/c ell | 1.01 RLU/min/c ell | 1.34 × 10−2 RLU/min/c ell | 2.0 × 10−4 RFU/min/c ell | 1.0 × 10−4 RFU/min/cell |
|
| ||||||
| DPI IC50 (nM) | 233 ± 122 | 180 ± 36 | 105 ± 55 | 42 ± 27 | 880 ± 214 | 6,488 ± 1,626 |
| 104 IC50 (nM) | 70 ± 57 | 481 ± 44 | 127 ± 20 | 44 ± 21 | 161 ± 153 | 4,136 ± 1,724 |
| 140 IC50 (nM) | 249 ± 116 | 177 ± 60 | 71 ± 15 | 55 ± 20 | 127 ± 110 | 1,990 ± 1,690 |
| 392 IC50 (nM) | 357 ± 268 | 850 ± 197 | 310 ± 53 | 208 ± 129 | 139 ± 143 | 225 ± 24 |
| 428 IC50 (nM) | 136 ± 81 | 324 ± 144 | 57 ± 35 | 35 ± 21 | 260 ± 254 | 412 ± 236 |
| 521 IC50 (nM) | 1,019 ± 361 | 1049 ± 126 | 1,470 ± 308 | 229 ± 123 | 5,533 ± 2,681 | 773 ± 362 |
PMN, polymorphonuclear leukocytes.