促红细胞生成素对早期外伤性癫大鼠的保护作用及对bcl-2和caspase-10表达的影响

Protective effect of erythropoietin on the early post-traumatic epilepsy rats and its effects on the expression of bcl-2 and caspase-10

目的研究促红细胞生成素(erythropoietin,EPO)对早期外伤性癫(post-traumatic epilepsy,PTE)大鼠的保护作用,及对bcl-2、caspase-10在皮层脑组织中表达的影响。方法选择30只成年SD大鼠作为研究对象,随机抽样原则分成实验组、模型组以及对照组。对照组大鼠仅予左侧感觉-运动皮层注射5μL生理盐水;模型组和实验组大鼠在相同位置注射等量的100mmol/L Fe Cl3,30min后实验组大鼠腹腔注射r Hu EPO 5000U/kg,模型组大鼠注射等量的生理盐水。观察行为学变化。24h后通过实时荧光定量PCR和Western blotting分别检测大鼠伤侧皮层脑组织中bcl-2、caspase-10的m RNA和蛋白表达水平。结果 (1)行为学:对照组未出现,实验组大鼠频率及发作时间均较模型组减少(P<0.05)。(2)RT-PCR检测:模型组和实验组bcl-2的m RNA表达均较对照组下降,实验组较模型组增高(P<0.05);模型组和实验组caspase-10的m RNA表达均较对照组增高,实验组较模型组下降(P<0.05)。(3)Western blotting检测:模型组和实验组bcl-2的蛋白表达均较对照组下降(P<0.05),实验组bcl-2的蛋白表达较模型组增高(P<0.05);模型组和实验组caspase-10的蛋白表达均较对照组增高,实验组caspase-10的蛋白表达较模型组下降(P<0.05)。结论铁离子致大鼠皮层脑组织中的bcl-2表达下降、caspase-10表达升高;EPO可上调bcl-2的表达水平、下调caspase-10的表达水平,抑制神经细胞凋亡,减轻铁离子致伤大鼠性发作,对早期PTE的脑损伤具有保护作用。

Objective To investigate the effects of erythropoietin on the expression of B cell lymphoma-2（ bcl-2） and cysteinylaspartate specific proteinase-10（ caspase-10） in cortical brain tissue,and to explore the protective effect on the early post-traumatic epilepsy rats. Methods Thirty adult rats were randomly divided into 3 groups. Rats in the control group were injected with 5 μL normal saline only in the left sensori-motor cortex; rats in the model group and experimental group were injected with the same amount of 100 mmol/L Fe Cl 3 at the same position. After 30 min,the rats in the experimental group were injected with r Hu EPO 5000 U/kg. The rats in the model group were injected with equal volume of saline. Changes in behavior were observed. After 24 hours,the m RNA and protein expression of bcl-2 and caspase-10 in rat cortical brain tissue were detected by real-time fluorescence quantitative PCR and Western blotting. Results（ 1） Behavior： there was no epileptic seizure in the control group. The seizure frequency and attack time in the experimental group was lower than those in the model group（ P 0. 05）.（ 2） RT-PCR： bcl-2 m RNA expression in the model group and the experimental group was lower than in the control group,which was higher in the experimental group than in the model group（ P 0. 05）; caspase-10 m RNA expression in the model group and the experimental group was higher than the control group,which was lower in the experimental group than the model group（ P 0. 05）.（ 3） Western blotting detection： the protein expression of bcl-2 in the model group and the experimental group was lower than that in the control group（ P 0. 05）. The expression of bcl-2 protein in the experimental group was higher than that in the model group（ P 0. 05）. The protein expression of caspase-10 in the model group and the experimental group was higher than that in the control group. The expression of caspase-10 protein in the experimental group was lower than that in the model group（ P 0. 05）. Conclusion The expression of bcl-2 is decreased and the expression of caspase-10 is increased in the cortical brain tissues of epileptic rats. EPO has a certain protective effect on the brain of iron-induced epilepsy rats through increasing the expression level of bcl-2 and reducing the expression level of caspase-10 in the cerebral cortex and reducing the frequency and degree of epileptic seizure.