| Objective: epilepsy is a common disease, and epidemiological investigation shows that the morbidity rate is 5‰ ~7‰. Seizures could cause not only necrosis or apoptosis of brain neurons as well as change of neurobiology, but also serious psychological disorder to patients and their relations. Status epilepticus(Status epilepticus, SE) is a common clinical critical disease, often resulting in the injury of hippocampus. If SE was not relieved and treated timely, the mortality rate would be high. The study found that the Puma((P53 up-regulated modulator of apoptosis, Puma) apoptosis protein had only a BH3 domain which played an important role in the process of apoptosis. At the same time, if the outer mitochondrial membrane permeability increased, cytochrome C(CytC)could be released and involved in endogenous mitochondrial apoptosis pathway. The damage mechanism of Puma and Cyt C was less study after SE. The experiment used SE rats model which was induced by pilocarpine to study the expression of Puma and CytC after SE 24 hours.Methods:The 62 adult male Sprague-Dawley(SD) rats were randomly divided into the epilepsy group(n=54) and the control group(n=8).The epilepsy group first application of atropine sulfate can effect to antagonize the effect of peripheral choline caused by pilocarpine. Then by intraperitoneal injection of pilocarpine to induce SE model of rats, according to the results, the epilepsy group was divided into status epilepticus group(SE group) and non status epilepticus group(non SE group).Hippocampus tissues of the rats were removed 24 h after SE. The brain tissues were stained by HE to observe the number of neurons in CA1 area of hippocampus, and were stained by TUNEL to survey hippocampus neurons injury and necrosis. The expression of Puma in hippocampus tissues was tested by Western blot and Cyt C was determined by Immunohistochemical. Pearson correlation analysis was adopted to explore the correlation between the expression of Puma and CytC. The control group except pilocarpine with physiological saline instead, and the rest did the same with experimental group.Results: There were 12 rats not induced to SE from the 54 epilepsy group, placed in the non SE group(n=12). There were 42 rats developed to SE in epilepsy group(n=54),the successful rate was 77.8%. And 14 rats died from the 42 SE rats within 24 h, so the mortality was 33.3%. The remaining rats(n=28) survived within 24 hours,and they were classified as SE group(n=28). The control group and non SE group was not death within24 hours. Compared to control group( 103.75±2.12) and non-SE group(100.67±2.19),the numbers of CA1 neurons stained by HE in SE group(54.75±3.37)were significantly decreased(p<0.01). Compared to control group(3.25±2.19) and non-SE group(3.33±2.02),the numbers of CA1 positive neurons stained by TUNEL in SE group(25.14±2.27) were significantly increased(p<0.05).And the number of CytC positive neurons in SE group(14.11±2.38)increased significantly(p<0.05) compared with control group(2.88±2.70)and non SE group(4.42±2.19). Moreover, the expression of Puma in SE group(0.439±0.038)increased significantly(p<0.05) compared with control group(0.016±0.003)and non SE group(0.249±0.034). Linear correlation analysis showed that there was a positive correlation between the expression of Puma and tunel positive neurons in SE group.(r=0.891,p<0.01).Moreover, there was a positive correlation between the expression of Puma and Cyt C in SE group.(r=0.895,P<0.01).Conclusion: There was neuronal apoptosis in hippocampus of rats after SE 24 h and there was a positive correlation between Puma and Cyt C. The expression of Puma and Cyt C increased in hippocampus neurons of rats in SE group, and there was a positive correlation between them. |
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