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Objective@#To explore the effects of n-butylphthalide (NBP) on mitochondria in hippocampus and learning and memory abilities in rats with chronic alcoholism.@*Methods@#60 male SD rats were randomly divided into three groups on average, including normal group, model group and treatment group, with 20 rats in each group.Rats of model group and treatment group are given 6% (V/V) alcohol solution continuously for 28 d to establish the model of chronic alcoholism.Rats in the treatment group were given butylphthalide for 14 days from the fourteenth day after giving alcohol solution.The Y type electric maze was used to test the learning and memory ability of rats, the content of H2S in the hippocampus and the activity of mitochondrial ATP enzyme were measured by spectrophotometry, and the protein expression of F-actin was detected by Western blot.@*Results@#Compared with the normal group, the learning and memory ability of the rats in the model group were decreased, the content of H2S in the hippocampus were increased, and the activity of mitochondrial ATP enzyme and the expression of F-actin protein were decreased, and most of the mitochondria were damaged under the electron microscope.The training times of the rats in treatment group(61.88±3.61)was lower than that of the model group(82.19±4.87), the ability of learning and memory was improved(P<0.05). Compared with the model group ((1.50±0.07)U/mgprot, (0.08±0.01)), the activity of the mitochondrial ATP enzyme((1.84±0.11)U/mgprot) and the level of F-actin protein(0.12±0.01)in rat hippocampus of treatment group were increased, the difference was statistically significant(both P<0.05). The level of H2S in rat hippocampus of the treatment group ((34.56±2.47) nmol/g) was lower than that of the model group ((44.55±3.71) nmol/g), the difference was statistically significant(P<0.05). Compared with model group, the mitochondrial damage of the hippocampus in the treatment group was improved under electron microscope.@*Conclusion@#NBP can abate mitochondrial damage and improve learning and memory abilities in chronic alcoholism rats.
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Objective To observe the changes of N-methyl-D-aspartate (NMDA) receptor 2B subunit (NR2B) expression in the striatum of chronic alcohol exposured rats at different withdrawal time.Methods 72 male Wistar rats were randomly divided into withdrawal 2h group,withdrawal 6h group,withdrawal 12h group,withdrawal 1d group,withdrawal 3d group and control group,and 12 rats in each group.In the 5 withdrawal groups,ethanol was administered in drinking water at the concentration of 6％ (V/V) for 16 weeks,and rats in control group were maintained with water.After 16 weeks ethanol was removed and ethanol withdrawal syndromes were evaluated.The expression of NR2B protein in the striatum was measured by immunofluorescence and western blot and the expression of NR2B mRNA in the striatum was measured by realtime PCR.Results Compared with withdrawal scores of control group((1.50±0.80)),scores of withdrawal 2h,6h,12h,1d,3d groups ((10.42±2.50),(15.42± 1.93),(9.25±2.01),(7.67± 1.92),(2.25±0.87) respectively) were higher,and the withdrawal scores of withdrawal 6h group were the highest.Compared with the expression of NR2B fluorescence intensity (2210.00± 178.20),the expression of NR2B protein(0.150±0.009) and the expression of NR2B mRNA(0.006±0.001) in the striatum of control group,the expression of NR2B fluorescence intensity (2710.56 ± 194.21),(5035.16 ± 234.41),(3326.23 ± 378.16),(2570.64 ±177.88),the expression of NR2B protein (0.192±0.008),(1.649±0.205),(0.783±0.109),(0.180±0.009) and the expression of NR2B mRNA (0.026±0.002),(0.351±0.034),(0.248± 0.023),(0.024±0.003) of withdrawal 2h,6h,12h,ld groups were significantly higher (P＜0.05),and with the extension of the withdrawal time,the expression was gradually increased.The expression of withdrawal 6h group was the highest,then began to decline,and returned to baseline levels at withdrawal 3 d(P＞0.05).Withdrawal scores were positively correlated with the expression of NR2B protein(r=0.719,P＜0.01),the expression of NR2B protein was positively correlated with the expression of NR2B mRNA(r=0.937,P＜0.01),and the expression of NR2B mRNA was positively correlated with withdrawal scores(r=0.673,P＜0.01).Conclusion The expression of NR2B was up-regulated in the striatum of chronic alcohol exposured rats at different withdrawl time.NR2B protein and NR2B mRNA expression is positively correlated with the withdrawal scores,suggesting that regulating the expression of NR2B may be a new target for the treatment of ethanol withdrawal symptoms.
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ObjectiveTo observe the effect of alcohol exposure during pregnancy on learning and memory and content of hydrogen sulfide(H2S) in the hippocampus of infant rats.MethodsThe animal models of alcohol exposure during pregnancy were made,and the learning and memory were evaluated by Y-maze in adult offspring.Content of H2S and activity of cystathionine-beta-synthase(GBS) in the hippocampus of the brain were evaluated with spectrophotometry;and CBS protein expression in the hippocampus was detected by immunohistochemistry.ResultsThe learning and memory ( (43.00 ± 15.33 ) times) of alcohol exposure during pregnancy group was significantly decreased compared with that of control and drinking groups (( 25.13 ± 12.35 )times and (26.12 ±11.95 ) times,P ＜ 0.05 ) ; spectrophotometry results showed that the content of H2S ( ( 30.32 ± 5.84 ) nmoL/g) of alcohol exposure during pregnancy group was significantly increased compared with that of control ( ( 52.51 ±7.85 ) nmol/g) and drinking groups( (49.93 ± 4.29 ) nmol/g),and the activity of CBS( ( 55.13 ± 4.45 ) nmol/g)of alcohol exposure during pregnancy group was significantly increased (P ＜ 0.01 ) compared with that of control ( (71.06 ± 5.58 ) nmol/g) and drinking groups( (69.96 ± 6.13 ) nmol/g) ; immunohistochemistry showed that the expressionof CBSproteinofalcoholexposureduringpregnancygroupwassignificantlyincreased.ConclusionThe damage effect of alcohol exposure during pregnancy on nerve system of infant rats may interrelate with down-regulation of H2S/CBS in the hippocampus.
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Objective To investigate the effect of butylphthalide (NBP) on H2S content and the expression of NR2B in the hippocampus of alcohol dependence rats. Methods A total of 84 SD male rats were randomly divided into 6 groups. Except for the normal group, other groups were subjected to alcohol solution with concentration of 6% ( V/V) for 28 d. Drug intervention began at the 14th day,and rats in the low,medium,high dose group were treated with NBP with a different concentration. Erden abstinence scoring was used to evaluate the rats withdrawal symptom. H2S content was measured in one side of hippocampus and CBS activity was tested in the other side of hippocampus. Hippocampus of 3 rats from each group was used to investigate NR2B mRNA level. Results Withdrawal symptom score ( 12.27 ± 1. 19),H2S content(30. 25 ±8.82), CBS activity (72. 44 ±7. 46) and NR2B mRNA expression( 19. 47 ±0. 86) in medium dose NBP group rats were lower than withdrawal symptom score(14.09 ±2.21) ,H2S content(44. 50 ±6. 65) , CBS activity(79. 06 ±4. 57) and NR2B mRNA expression (29. 13 ±1.39) in experimental control group (P<0.05). Withdrawal symptom score(12. 18 ±1.08) ,H2S content(33.00 ±5.38) ,CBS activity(67. 81 ±9. 37) and NR2B mRNA expression(23. 12 ± 1. 86) in high dose NBP group rats were lower than experimental control group (P < 0. 05). Conclusion NBP can reduce withdrawal symptoms of alcohol dependence rats,may be related to decreased expression of H2S/CBS system, and NR2B mRNA expression.
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AIM:In order to investigate the molecular mechanism of alcoholism acting on learning and memory,the dysfunction of learning and memory function was observed and the content of nitric oxide(NO)and neuronal nitric oxide synthase(nNOS)were determined in rats with acute alcoholism.METHODS:The mature male Sprague-Dawley rats were randomly divided into two groups.The experimental group animals were intraperitoneally administered with ethanol.The control group animals were injected with saline in the same way.The tests of learning and memory were performed at Y-maze after 6 h.Then brains were removed and the content of NO in brain tissue and nNOS expression in hippocampus CA1,corpus striatum were determined,respectively.RESULTS:(1)The training times to reach qualifying standards of Y-maze in experimental group(34.33?13.04)were higher than those in control group(27.50?8.79,P