Table 3.
Effect of 24 h of hypoxia on caspase 3 activity and ATP levels in pcDNA3.1 and pcDNA3.1 TSPO-transfected LLC-PK1 cells
|
Hypoxia |
||
|---|---|---|
| Caspase 3 Activity, % | ATP Level, % | |
| EV (24 h) | 100 | 18.21±1.1 |
| TSPO (24 h) | 37±6.2* | 41.21±17.9* |
| EV+PK11195 (24 h) | 97±5.6/96±6.3/95 | 19.30±1.4/18.25±1.6/8.80±2.1† |
| 1 nM/10 nM/100 nM | ||
| TSPO+PK11195 (24 h) | 35±4.6*/37±3.8*/38±4* | 43.11±14.2*/45±12*/46±11* |
| 1 nM/10 nM/100 nM | ||
| EV+Ro5-4864 (24 h) | 99±6.2/98±5.1/97±4.9 | 15.31±1.3/16.3±1.8/8.7±2.3† |
| 1 nM/10 nM/100 nM | ||
| TSPO EV+Ro5-4864 (24 h) | 41±6.4*/43±3.7*/46±4.1* | 41.21±17.9*/44.2±14*/45.3±13* |
| 1 nM/10 nM/100 nM | ||
| EV+clonazepam (24 h) | 96±5.8 | 16.66±1.8 |
| TSPO+clonazepam (24 h) | 38±5.3*/44±4.2*/45±3.9 | 42.3±13.1*/43.1±11*/43.8±16.3* |
| Reoygenation | ||
| EV (24 h) | 100 | 13±0,9 |
| TSPO (24 h) | 24±4,1* | 53±11.8* |
| EV+PK11195 (24 h) | 91±3.3/92±5.1/91±3.7 | 14.10±1.1/13.4±1.9/7.2±3.7 |
| 1 nM/10 nM/100 nM | ||
| TSPO+PK11195 (24 h) | 25±3.8*/17±1.2*/11±3.7* | 54.1±12.2*/65±9.4*/77±12.8* |
| 1 nM/10 nM/100 nM | ||
| EV+Ro5-4864 (24 h) | 90±3.4/92±4.3/93±3;3 | 12.5±2.3/11.53±2.5/6.3±2.0 |
| 1 nM/10 nM/100 nM | ||
| TSPO EV+Ro5-4864 (24 h) | 21±3.4*/12±1.7*/7±1.1 | 51.4±11.9*/69.2±10*/75.3±11* |
| 1 nM/10 nM/100 nM | ||
| EV+clonazepam (24 h) | 90±3.6 | 15.7±2.9 |
| TSPO+clonazepam (24 h) | 28±3.6*/24±3.8*/25±5.6 | 55±10.3*/56.5±10*/58.9±11.6* |
Values are means ± SE of 6 independent experiments. Caspase 3 activity results are expressed as percentage of activity at 24 h of hypoxia and ATP results as percentage between values measured at time 0 and after 24 h of hypoxia and 6 h of reoxygenation. PK 11195 and RO5-4864 were used at 3 different concentrations: 1, 10, 100 nM (left to right in the results). Clonazepam was used at 5 nM. EV, cells with empty virus; TSPO, cells which overexpressed TSPO.
*
P < 0.05 TSPO-transfected LLC-PK1 cells vs. LLC-PK1 with empty virus.
†
P <0.05 100 nM dose vs. 1 and 10 nM doses.