Animals and drug treatment Male or female Sprague–Dawley rats (180 to 230 g) were employed for the experiments (Shanghai Experimental Animal Center, Chinese Academy of Sciences). Five rats were kept in individual cages with water and food available ad libitum. The animal room
was maintained at 21°C to 23°C, with a 12-h light–dark cycle. All experimental procedures were approved by the Committee of Laboratory Animals, Chinese Academy of Sciences. Rats were intraperitoneally (i.p.) administered with 70-mg/kg dose of 1% PTZ (dissolved in saline) to induced auditory evoked potential (AEP). Control animals received the same amount of saline injections. The seizures were rated according to the following criteria [34, 35]: stage 0, no VX-770 in vitro response; stage I, ear and facial
twitching; stage II, myoclonic jerks without upright position; stage III, myoclonic jerks, upright position with bilateral forelimb clonus; stage IV, clonic-tonic seizure; and stage V, generalized clonic-tonic seizures, loss of postural control. Experimental rats were divided into four groups as follows: group 1, rats were treated with saline; group 2, rats were i.p. injected with a dose of 70 mg/kg PTZ to induce the onset of seizures; group 3, rats were i.p. co-administered with a dose of 70 mg/kg PTZ since i.p. injected with a dose of 500 mg/kg taurine after 30 min; and group 4, rats were i.p. co-administered with a dose of 70 mg/kg PTZ since i.p. injected with a dose of 500 mg/kg GABA after 30 min. After 1 h, the animals were killed, the brains were dissected, SRT2104 order the cerebral cortex and hippocampus tissues were removed, and blood was withdrawn. The brain tissue was rinsed in ice-cold normal saline, added to nine times ice-cold normal saline, homogenized, and centrifuged at 5,000×g for 15 min at 4°C. The blood nearly was centrifuged at 3,000×g for 15 min. The supernatant and serum were obtained and stored in a −20°C refrigerator for MDA Blasticidin S research buy assays and antioxidant enzymes’ (SOD, GSH-Px) activity assays. The
protein concentration was determined by Coomassie Brilliant Blue method. MDA assay and antioxidant enzyme activity measurement The MDA and antioxidant enzymes’ (SOD, GSH-Px) activity of the cerebral cortex and the hippocampus tissue and blood from PTZ-induced AEP were evaluated by MDA assay and antioxidant enzymes’ (SOD, GSH-Px) kits according to the manufacturer’s instructions. Statistics Data were shown as mean ± S.E.M. Statistical evaluation was carried out by one-way analysis of variance (ANOVA) followed by Scheffe’s multiple range tests. P < 0.05 was considered to be significant. Results Incubation products assayed by HPLC and fluorescence The mixture was separated at acidic pH through HPLC and fluorescence after amino acids (5.0 mM) were incubated with MDA (5.0 mM) in 0.2 M PBS, pH 7.4, at 37°C for 48 h.