Effects of macromolecular synthesis inhibitors, an endonuclease inhibitor, and antioxidants on apoptotic cell death induced by oxidative insults and HNE in PC12 cells
| Insult | Vehicle | Cells with apoptotic nuclei (%) | ||||
|---|---|---|---|---|---|---|
| Cyclohex | Act-D | ATA | VitE | PG | ||
| Cont (2 hr) | 3 ± 1.2 | 1 ± 0.3 | 2 ± 0.6 | 1 ± 0.3 | 0 ± 0.3 | 1 ± 0.3 |
| Cont (72 hr) | 24 ± 4.7 | 11 ± 2.5 | 10 ± 3.3 | 9 ± 2.4 | 10 ± 2.1 | 13 ± 3.1 |
| FeSO4 | 75 ± 3.2* | 16 ± 2.71-160 | 22 ± 3.31-160 | 13 ± 3.31-160 | 13 ± 2.81-160 | 21 ± 2.11-160 |
| Aβ | 72 ± 3.3* | 21 ± 3.41-160 | 23 ± 4.31-160 | 12 ± 2.21-160 | 11 ± 3.51-160 | 14 ± 3.21-160 |
| HNE | 71 ± 6.7* | 29 ± 4.31-160 | 29 ± 4.01-160 | 26 ± 3.21-160 | 59 ± 3.3 | 65 ± 3.8 |
-
Cultures were pretreated for 2 hr with the indicated agents: 0.2% ethanol (Vehicle), 10 μm cycloheximide (Cyclohex), 5 μm actinomycin-D (Act-D), 100 μmaurintricarboxylic acid (ATA), 50 μg/ml vitamin E (VitE), 10 μm propyl gallate (PG), or 1 mmglutathione-ethyl ester (GSH). Some cultures were fixed at that point (Cont, 2 hr), while parallel cultures were exposed for 72 hr to 0.2% ethanol (Cont 72 hr), 1 mm FeSO4, 50 μm Aβ, or 10 μm HNE. Cells then were fixed and stained with Hoescht dye, and percentages of cells exhibiting nuclear condensation and fragmentation were determined. Values are the mean and SEM of determinations made in three or four separate cultures.
-
↵* p < 0.001 compared with value for control vehicle-treated (72 hr) cultures;
-
↵F1-160 p < 0.01 compared with value for vehicle-treated cultures exposed to the same insult. ANOVA with Scheffé’s post hoc tests. Preliminary studies showed that 10 μm cycloheximide reduced levels of protein synthesis by >90% during a 24 hr exposure period (data not shown).
