癫痫大鼠海马硫氧还蛋白还原酶活性及cleaved caspase-3表达检测

Detection of thioredoxin reductase activity and the expression of cleaved caspase-3 in hippocampus of seizure rats

目的研究海人酸诱导癫痫大鼠海马组织硫氧还蛋白还原酶(TrxR)活性及cleaved caspase-3表达变化。方法 Sprague-Dawley大鼠随机分为癫痫对照组(NS组)和癫痫组(KA组)。采用脑室注射海人酸制作大鼠癫痫模型。海人酸注射6h取海马组织匀浆,应用5,5-二硫代双(2-硝基苯甲酸)还原法对海马TrxR活性进行测定,采用western blotting法检测cleaved caspase-3表达。结果 NS组、KA组TrxR活性分别为:(71±19)、(116±40)U/mg;KA组cleaved caspase-3的表达较NS组明显增加,差异有统计学意义(P<0.05)。结论癫痫大鼠海马组织TrxR活性明显增加,引起caspase-3裂解,导致神经元凋亡。

Objective To detect and analyze the thioredoxin reductase（TrxR） activity and the expression of cleaved caspase-3 in hippocampus of seizure rats induced by kainic acid（KA）.Methods Adult male Sprague-Daley rats were allotted into 2 groups as followed ：control group（NS） and seizure group（KA）.Seizures were induced by intracerebroventricular injection of KA dissolved in sterile saline.TrxR activity was detected by dithio-bis-nitrobenzoic acid reduction assay.Western blotting was used to analyze the expression of cleaved caspase-3.Results Thioredoxin reductase activity were（71±19）U/mg,（116±40）U/mg respectively.Compared with the control group,the expression of cleaved caspase-3 increased significantly in the seizure group（P0.05）.Conclusion TrxR activity increases in hippocampus of seizure rats induced by KA,subsequently activates caspase-3 and ultimately results in neuronal apoptosis.