| Objective: Carbamazepine(CBZ), as a tricyclic anticonvulsant,is mainly used for clinical anti-convulsant and anti-epileptic. It is one of the first choice for patients with phychomotor seizures, and CBZ can also be used to treat trigeminal neuralgia, glossopharyngeal neuralgia and manic-depression and so on. Because of its wider clinical application,adverse reactions to multiple systems are also receiving close attention increasingly. Adverse effects to the nervous system,such as dizziness, ataxia, drowsiness,impulsive behavior and hallucinations frequently reported in the literature. This experiment is with the purpose of studing the correlation of long-term administration of CBZ to rats and the apoptosis in neurons of these rats, so that anlyse some of the Adverse effects of CBZ from the point of view of animal experiments accodingly,as well as providing a further study of CBZ experimental evidence and experimental direction.Methods:1 Grouping and drug intervention: 30 healthy Sprague-Dawley rats aged ten weeks (weighted 200±20g) were divided into six groups randomly n = 5. Grouped as follows:Group A: Long course high-dose CBZ group, rats were treated with CBZ by 500mg/kg/d deliquescing in normal saline (2ml/100g) for 30 days;Group B: Long course low-dose CBZ group, rats were treated with CBZ by 125mg/kg/d deliquescing in normal saline (2ml/100g) for 30 days;Group C: Long course normal saline group, rats were treated with normal saline (2ml/100g) for 30 days;Group D: Short course high-dose CBZ group, rats were treated with CBZ by 500mg/kg/d deliquescing in normal saline (2ml/100g) for 15 days;Group E: Short course low-dose CBZ group, rats were treated with CBZ by 125mg/kg/d deliquescing in normal saline (2ml/100g) for 15 days;Group F: Short course normal saline group, rats were treated with normal saline (2ml/100g) for 15 days.2 Preparation of samples: Infuse rats with 4% glutaraldehyde from the left ventricle for about half an hour after successfull anesthesia with 4% chloral hydrate. Cut down the frontal lobe and cerebellar of rats and put them into 4% paraformaldehyde devidedly. Conventional gradient dehydration, pellucidum, dipping wax and embedding.3 Apoptosis detection: Select frontal lobe and cerebellum then give them conventional continuous coronal slices, slice thickness of 5μm. Detect the apoptotic neurons in the samples in by TUNEL and the expression of apoptosis-related genes bax and bcl-2.4 Statics: Use One-Way ANOVA to compare the differences between groups, and the comparison between any two groups was achieved by SNK method. P=0.05 as the check standard.Results:1 The comparison of apoptosis in every group after drug intervention in frontal lobe:1.1 Postive neurons detected by TUNEL:thin and scattered TUNEL-positive neurons could be seen in every groups. Comparing the mean of every two groups got the result of P>0.05,The differences were not statistically significant.1.2 Detecting the positive expression of apoptosis-related genes bax and bcl-2 by immunohistochemisty: neurons with positive expression of bax and bcl-2 could be seen in a small amount in every group, expressed as the nucleus and cytoplasm brown. Statisticsed the mean of positive neurons in each group,in any two groups P>0.05, so the differences were not statistically significant. Calculated the radio of positive expression of bax and bcl-2 bax/bcl-2 and statisticsed, in any two groups P>0.05, and the differences were not statistically significant.2 The comparison of apoptosis in every group after drug intervention in cerebellar:2.1 Postive neurons detected by TUNEL: TUNEL-positive neurons increased visibility in Group A, and some were patchy distribution. More TUNEL-positive neurons were also visible in Group D. In the other groups scattered TUNEL-positive neurons could be seen. Comparing Group A and Group C, P<0.01, there was significant difference between the two; Comparing Group B and Group C, P> 0.05, the difference between the two was not significant;Comparing Group D and Group F, P <0.05, there was significant difference between the two; Comparing Group E and Group F, P> 0.05, the difference between the two was not significant;Comparing Group A and Group D, P <0.05, there was significant difference between the two.2.2 Detecting the positive expression of apoptosis-related genes bax and bcl-2 by immunohistochemisty: increased significantly Bax positive neurons could be seen in Group A, more Bax positive neurons were also visible in Group D. Both Bax and Bcl-2 positive neurons were sparse and scattered in other groups. Statistical analysis for Bax positive neurons and Bax/Bcl-2 both showed that: comparing Group A and Group C, P<0.01, there was significant difference between the two; Comparing Group B and Group C, P> 0.05,the difference between the two was not significant; Comparing Group D and Group F,P <0.05, there was significant difference between the two; Comparing Group E and Group F, P> 0.05, the difference between the two was not significant; Comparing Group A and Group D, P <0.05, there was significant difference between the two.Conclusions:1 Fedding adult rats with CBZ do not cause the neurons excessive apoptosis in frontal lobe;2 Fedding adult rats with high dose CBZ can lead to the positive expression of bax in the neurons of cerebellum increasing significantly. It suggests that CBZ may promote apoptosis in neuron by increasing the expression of bax; and the degree of apoptosis is gradually increasing with administration time;3 Fedding adult rats with low dose CBZ do not lead to excessive apoptosis in neurons in cerebellum.
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