Abstract
Purpose: To investigate the neuroprotective effect of sulfanilamide benzotriazole tetrazole (SBT) in neonatal rats exposed to isoflurane, and also to elucidate the underlying mechanism.
 Methods: Rat pups (n = 60) were randomly assigned to six groups of 10 pups each: normal control group, negative control group, 5 mg/kg SBT group, 10 mg/kg SBT group, 15 mg/kg SBT group, and 20 mg/kg SBT group. With exception of normal control group, pups were exposed to isoflurane (0.75 %) for 6 h on postnatal day 7. The negative control group was not treated, while pups in the four treatment groups received 5, 10, 15 and 20 mg/kg SBT, respectively, 1 h after exposure to anaesthesia. TUNEL assay was used to determine the extent of apoptosis in cornu ammonis area-1 (CA-1), cornu ammonis area-3 (CA-3) and dentate gyrus of rat hippocampal tissues. Expressions of apoptotic and anti-apoptotic proteins were determined using Western blotting. Evaluation of learning and memorizing ability was done using Morris water maze test.
 Results: Isoflurane significantly increased the extent of apoptosis in CA-1, CA-3 and dentate gyrus of rat hippocampal tissues (p < 0.05). However, treatment with SBT significantly and dose-dependently reduced neuronal apoptosis (p < 0.05). The expression of caspase 3 was significantly upregulated by isoflurane, but was significantly and dose-dependently down-regulated by SBT (p < 0.05). Isoflurane significantly increased Bax expression, and decreased the expression of bcl-2 (p < 0.05). The effects of isoflurane on the expression of these proteins were significantly and dose-dependently reversed by SBT (p < 0.05). The expression of bcl xL in rat hippocampal tissues was significantly down-regulated by isoflurane, but was significantly and dose-dependently upregulated by SBT (p < 0.05). The escape latency of pups was significantly higher in negative control group than in normal control group, but SBT treatment significantly and dose-dependently reversed this trend (p < 0.05).
 Conclusion: These results suggest that SBT prevents neuronal apoptosis, and improves the ability to learn and memorize in neonatal rats exposed to isoflurane via regulation of apoptotic, JNK and p38 MAPK protein expressions.
Highlights
Tetrazoles are a class of synthetic organic heterocyclic compounds consisting of a 5-membered ring containing four nitrogen atoms and a carbon atom
The present study investigated the neuroprotective effect of sulfanilamide benzotriazole tetrazole (SBT) in neonatal rats exposed to isoflurane, and the underlying mechanism
Isoflurane significantly increased the extent of apoptosis in the cornu ammonis area-1 (CA-1), cornu ammonis area-3 (CA-3) and dentate gyrus of rat hippocampal tissues (p < 0.05)
Summary
Tetrazoles are a class of synthetic organic heterocyclic compounds consisting of a 5-. Rabbit polyclonal anticaspase-3, bcl-xL, bad, p-bad, bax, bcl-2, ERK1/2, p-ERK1/2, JNK, p-JNK, p-c-Jun, p38, p-p38, and β-actin were products of Abcam (USA), while ANY-maze was obtained from Stoelting Co., Ltd. Neuronal apoptosis and protein expression in pups hippocampal tissues were determined. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used for the determination of apoptosis in pups hippocampal tissues. The pups were put into the pool twice daily and allowed to locate the submerged platform within 60 sec. Impairment of cognitive behaviour was determined in the pups on postnatal day 30 using cued recall test. A rod placed around 20 cm above the water in any one of the 40 quadrants randomly served as a cue to enable the pups locate the platform. Pups were placed in the same position throughout the trials to determine the time taken to reach the platform. Values of p < 0.05 were considered statistically significant
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