匹罗卡品致大鼠癫痫持续状态后海马组织中Puma和CytC的表达变化

Changes of expressions of Puma and CytC in hippocampus tissues of rats with pilocarpine-induced status epilepticus

目的研究匹罗卡品致癫痫持续状态(SE)的大鼠模型中,P53上调细胞凋亡调控因子(Puma)和细胞色素C(Cyt C)在大鼠海马组织中的表达情况。方法 10%的匹罗卡品(300 mg/kg)腹腔注射于成年雄性SD大鼠,诱发SE 60 min后终止发作。24 h后取大鼠海马组织,苏木精-伊红(H-E)染色观察海马CA1区神经元的数量,TUNEL染色观察海马神经元的损伤情况。采用Western blotting方法测定海马组织中Puma的表达,免疫组织化学方法检测海马组织中Cyt C的含量。运用Pearson相关分析法分析Puma和Cyt C表达的相关性。结果 SE后24 h,大鼠海马CA1区H-E染色神经元数量显著减少,TUNEL阳性神经元含量明显增加。可见Puma的表达,并可见大量的Cyt C阳性神经元。且Puma和Cyt C的含量变化具有相关性(r=0.906,P=0.000)。结论大鼠SE后24 h海马神经元出现凋亡,Puma和Cyt C在SE大鼠海马组织神经元中的表达含量增加,两者的表达水平成正相关。

Objective To study the expressions of P53 up-regulated modulator of apoptosis（ Puma） and cytochrome C（ CytC） in hippocampus tissues of rats in pilocarpine-induced status epilepticus（ SE） rat model. Methods Adult male Sprague Dawley（ SD） rats were intraperitoneally injected with pilocarpine of 10%（ 300 mg / kg） and the seizure was ceased 60 min after SE was induced. Hippocampus tissues of the rats were removed after 24 h. Brain tissues were stained by H-E and the number of neurons in CA1 area of hippocampus was observed. Then brain tissues were stained by TUNEL and the injury of hippocampus neurons was observed. The Western blotting was adopted to detect the expression of Puma in hippocampus tissues and immunohistochemical method was used to detect the CytC level in hippocampus tissues. The correlation of expressions of Puma and CytC was analyzed by the Pearson correlation analysis. Results The number of stained neurons in the CA1 area of hippocampus greatly decreased 24 h after SE and the number of TUNEL positive neurons significantly increased. Expressions of Puma and many CytC positive neurons appeared. The expression of Puma was correlated with the expression of CytC（ r = 0. 906, P = 0. 000）.Conclusion Apoptosis is found in hippocampus neurons of rats 24 h after SE. Expressions of Puma and CytC increase in hippocampus neurons of rats with SE and the expression of Puma is positively correlated with the expression of CytC.