Table 3. Lysosomal membrane damage induced by sumatriptan in rat liver hepatocytes and protective effect of antioxidants, radical scavengers, lysosomal membrane stabilizers, mitochondrial ATP generators, MPT pore sealing agents and CYP450 inhibitors .
Treatment |
Acridine orange redistribution
(intensity unit of diffused cytosolic green fluorescence) |
||
Incubation time | |||
15 min | 30 min | 60 min | |
Control (intact hepatocytes) | 7.5 ± 2 | 9.9 ± 3 | 11.5 ± 3 |
Sumatriptan (5 mM) | 34.3 ± 2a | 39.5 ± 3a | 53.2 ± 3a |
+Taurine (200 µM) | 14.3 ± 2b | 17.5 ± 3b | 24.6 ± 2b |
+Quercetin (500 µM) | 17.1 ± 2b | 20.4 ± 2b | 28.6 ± 3b |
+NAC (200 µM) | 15.3 ± 2b | 18.6 ± 3b | 26.9 ± 3b |
+Vitamin E (100 µM) | 19.4± 2b | 24.8 ± 2b | 31.5 ± 1b |
+BHT (50 μM) | 19.3 ± 2b | 27.5 ± 1b | 39.1 ± 1b |
+Methylamine (30 mM) | 18.4 ± 2b | 21.1 ± 3b | 30.7 ± 1b |
+Chloroquine (100 μM) | 20.1 ± 2b | 22.2 ± 1b | 36.0 ± 1b |
+3-Methyladenine (5 mM) | 21.1 ± 3b | 25.5 ± 4b | 34.9 ± 3b |
+Fructose (10 mM) | 19.8 ± 2b | 27.3 ± 1b | 38.1 ± 2b |
+L- Gln (1 mM) | 18.3 ± 2b | 26.1 ± 2b | 38.7 ± 3b |
+ TFP (15 µM) | 19.2 ± 3b | 25.5 ± 3b | 32.2 ± 2b |
+Carnitine (2 mM) | 20.3 ± 3b | 24.9 ± 5b | 37.1 ± 3b |
+4- MP (500 µM) | 17.2 ± 2b | 22.9 ± 3b | 31.3 ± 4b |
+Cimetidine (2 mM) | 18.6 ± 2b | 24.7 ± 3b | 33.9 ± 3b |
GSH depleted hepatocytes | 59.8 ± 2 | 87.4 ± 2 | 121.2 ± 1 |
+ Sumatriptan (5 mM) (in GSH depleted hepatocytes) | 45.6 ± 3c | 53.2 ± 2c | 68.2 ± 2c |
Lysosomal membrane fragility was measured as fluorescent intensity unit of diffused cytosolic green fluorescence induced by acridine orange following the redistribution from lysosomes into cytosol in acridine orange loaded hepatocytes. Results are expressed as mean ±S.E. of three separate experiments (n=3).
a Significantly higher than control (p < 0.05).
bSignificantly lower than sumatriptan treated hepatocytes (p < 0.05).
cSignificantly higher than sumatriptan treated hepatocytes (p < 0.05).