ABSTRACT
Objective To evaluate the role of nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase in bupivacaine-induced injury to neurons.Methods SH-SY5Y cells were seeded in 96-well culture plates at a density of 5× 104 cells/well and randomly divided into 3 groups (n=24each) using a random number table:control group (group C),bupivacaine group (group B),and apocynin (NADPH oxidase inhibitor) + bupivacaine group (group A+B).The cells were cultured in a serum-free medium in group C.The cells were cultured in a serum-free medium containing 1 mmol/L bupivacaine in group B.In group A + B,the cells were cultured for 30 min in a serum-free medium containing apocynin 100 μmol/L,and then cultured in a serum-free medium containing 1 mmol/L bupivacaine.At 2,4 and 6 h of incubation,the cells in 6 wells of each group were selected to evaluate the cell viability by MTS assay.At 4 h of incubation,the cells in 6 wells of each group were selected to detect reactive oxygen species (ROS) level by flow cytometry.Results Compared with group C,the cell viability was significantly decreased at 4 and 6 h of incubation,and the production of ROS was increased at 4 h of incubation in group B (P＜ 0.05).Compared with group B,the cell viability was significantly increased at 4 and 6 h of incubation,and the production of ROS was decreased at 4 h of incubation in group B (P＜0.05).Conclusion NADPH oxidase is involved in bupivacaine-induced injury to neurons.
ABSTRACT
Objective To evaluate the effect of docosahexaenoic acid (DHA) on hepatic ischemia-reperfusion (I/R) injury in rats.Methods Fifteen male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were randomly divided into 3 groups (n =5 each) using a random number table: sham operation group (group S), hepatic I/R group (group I/R) , and group DHA.Hepatic I/R was induced by clamping the hepatic pedicle supplying the left and middle lobes of the liver for 60 min, followed by 24 h reperfusion in anesthetized rats.DHA 4 mg/kg was injected intravenously at 30 min before ischemia and 10 min of reperfusion in group DHA.The equal volume of solvent was given instead in S and I/R groups.Blood samples were taken from the inferior vena cava at 24 h of reperfusion for determination of serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities, and resolvin D1 concentrations.The rats were then sacrificed, and the livers were removed for determination of myeloperoxidase (MPO) activity (by spectrophotometry), and interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) mRNA expression (by quantitative real-time reverse transcriptase polymerase chain reaction).The livers were cut into sections which were stained with haematoxylin and eosin, and examined under light microscope.Results Compared to group S, the serum ALT and AST activities, serum resolvin D1 concentrations, and MPO activity, and IL-6 and TNF-α mRNA expression in liver tissues were significantly increased in I/R and DHA groups (P＜0.05).Compared to group Ⅰ/R, the serum resolvin 1D1 concentrations, and MPO activity and TNF-α mRNA expression in liver tissues were significantly decreased (P＜0.05) , and no significant difference was found in the serum ALT and AST activities in group DHA (P＞0.05).There was no significant difference in pathological changes of the liver between group DHA and group I/R.Conclusion DHA can attenuate inflammatory responses during hepatic I/R, but it is not sufficient to mitigate liver injury in rats.