| Objective: Carbamazepine (CBZ) is widely used for clinical aspects. It isone of the main drugs in the treatment seizures and all kinds of peripheralneuropathy, and it can also be used to antimanic-depression, arrhythmia,antidiuretic, to treat tinnitus, facial spasms and alcohol withdrawal syndrome,etc. Because of its broad-spectrum, effective, low price, carbamazepinecurrently applied widely, but at the same time the reports of serious adversereactions to increasing, also is the attention of people. Carbamazepine adversereactions caused by involving multiple systems in the body, including skin andaccessories, central nervous system, the digestive system, blood andcardiovascular system, etc. The nervous system adverse reactions caused bycommon have vertigo, drowsiness, ataxia, hallucinations, memory loss, lack ofconcentration and disorder of consciousness and so on. At present, to reduceor avoid the occurrence of adverse reactions has become one of the mostconcerned issues in people. This experiment is to observe the effects ofcarbamazepine on apoptosis of neurons in the hippocampus and parietal cortexof rats, to discuss the possible mechanism of the occurrence of adversereactions, to offer theory according to avoid the side effects and direct doctorsin choosing the carbamazepine.Methods:1Experimental animals and groups:30SPF male Sprague-Dawley(SD) rats,weighing(200±20)g, were randomly divided into three groups, eachcomponent had2groups, which had five rats. Grouped as follows: N normalsaline group：①rats were treated with normal saline for15days②rats weretreated with normal saline for30days; L low-dose carbamazepine group:①rats were treated with low-dose carbamazepine for15days②rats weretreated with low-dose carbamazepine for15days; H high-dose carbamazepine group:①rats were treated with high-dose carbamazepine for15days②ratswere treated with high-dose carbamazepine for30days.2Drug administrations: Filling the stomach method, continuous irrigationstomach every day. As follows: low-dose carbamazepine group: rats wereirrigated with carbamazepine by125mg/kg/d deliquescing in normal saline(2ml/100g); high-dose carbamazepine group： rats were irrigated withcarbamazepine by500mg/kg/d deliquescing in normal saline (2ml/100g);normal saline group：rats were irrigated with equal dose of normal saline(2ml/100g/d).3Preparation of samples: After successful anesthesia with10%hydrationchlorine aldehyde were intraperitoneal injected, the rats were first infused thenormal saline from left ventricular, and then infused the4%paraformaldehydefirst after fast slow. Then cut down the hippocampus and parietal cortex of ratsquickly and put them into4%paraformaldehyde, Stepwise ethanoldehydration, Xylene transparent, Paraffin embedding.4Samples detection: Selected the hippocampus and parietal cortex then gavethem conventional continuous coronal slices, slice thickness of5μm. Thespecimens were detected by three methods: observed the morphologicalcharacteristics of the nerve cell by HE staining in light microscope; detectedthe apoptotic neurons in the samples by TUNEL; examined the expression ofapoptosis-related genes fas and caspase-3by immunohistochemical SPmethod.5Statics: Use the analysis of variance in the statistical analysis softwareSPSS13.0, with P <0.05for the check standards, think there is statisticalsignificance. Use the One-Way ANOVA to compare the differences betweengroups, and the comparison between any two groups was achieved by SNKmethod.Results:1Results of HE staining:The nerve cells in the hippocampus from the carbamazepine groups wasnot clear, the distance between cells became larger, arranged messy, hyperchromatic nuclei became pyknotic, visible to the empty bubble formedaround, and this phenomenon was especially serious in the high-dose groups.The cell in the normal saline groups arranged regularly, did not see theperformance.The nerve cells in the parietal cortex from the carbamazepine groupsarranged regularly, some time could be seen the cells shrinkage and the emptybubble formed around, and there was no obvious change with the parietalcortex form the normal saline groups.2Results of the TUNEL:More TUNEL-positive cells could be seen in the hippocampus of eachcarbamazepine group. TUNEL-positive neurons increased visibility in thehigh-dose groups (Group H1and Group H2) and were patchy distribution. Inthe other two groups scattered TUNEL-positive neurons could be seen.Comparing Group H2and Group N2, there was significant difference betweenthe two(P<0.05); Comparing Group H1and Group N1, there was significantdifference between the two(P<0.05); Comparing Group L2and Group N2, thedifference between the two was not significant(P>0.05); Comparing Group L1and Group N1, the difference between the two was not significant(P>0.05);Comparing Group H2and Group H1, there was significant difference betweenthe two(P<0.05).Scattered TUNEL-positive neurons could be seen in the parietal cortex ofeach carbamazepine group. Comparing the mean of every two groups, thedifferences were not statistically significant(P>0.05).3Immunohistochemistry results:3.1Expression of fas:Neurons with positive expression of fas could be seen in the hippocampusof each carbamazepine group. Only a little could be seen in the normal salinegroups. The expression of fas increased visibility in the high-dose groups.Comparing the expression of Group H2and Group N2, there was significantdifference between the two(P<0.05); Comparing Group H1and Group N1,there was significant difference between the two(P<0.05); Comparing Group L2and Group N2, the difference between the two was not significant(P>0.05);Comparing Group L1and Group N1, the difference between the two was notsignificant(P>0.05); Comparing Group H2and Group H1, there was significantdifference between the two(P<0.05).Neurons with positive expression of fas could also be seen in the parietalcortex of each carbamazepine group, but the expressions were lower.Comparing the expression of every two groups, the differences were notstatistically significant(P>0.05).3.2Expression of capase-3:Neurons with positive expression of caspase-3could be seen in thehippocampus of each carbamazepine group. The expression of caspase-3increased visibility in the high-dose groups. Comparing the expression ofGroup H2and Group N2, there was significant difference between thetwo(P<0.05); Comparing Group H1and Group N1, there was significantdifference between the two(P<0.05); Comparing Group L2and Group N2, thedifference between the two was not significant(P>0.05); Comparing Group L1and Group N1, the difference between the two was not significant(P>0.05);Comparing Group H2and Group H1, there was significant difference betweenthe two(P<0.05).Neurons with positive expression of caspase-3could also be seen in theparietal cortex of each carbamazepine group, but the expressions were lower.Comparing the expression of every two groups, the differences were notstatistically significant(P>0.05).3.3The relationship between the expression of fas and caspase-3:The expression of fas and caspase-3were positively correlated in thehippocampus of each carbamazepine group.Conclusions:1Feeding rats with carbamazepine do not cause the neurons excessiveapoptosis in the parietal cortex;2Feeding rats with low-dose carbamazepine do not cause the neuronsexcessive apoptosis in the hippocampus; 3Feeding rats with high-dose carbamazepine can cause the cell excessiveapoptosis and lead to the positive expression of fas and caspase-3in theneurons of hippocampus increasing significantly, and the degree of apoptosisis gradually increasing with administration time;4The expression of fas and caspase-3were positively correlated in thehippocampus of each group. It suggests that carbamazepine may promoteapoptosis in the hippocampus neuron by the fas mediated apoptosis way. |
PDF Full Text Request