目的:探讨白藜芦醇(resveratrol,Res)在氯化锂(LiCl)-重复低剂量匹罗卡品(pilocarpine,Pilo)诱导癫痫持续状态(status epilepticus,SE)大鼠模型中的抗氧化作用及其机制。方法:54只SD大鼠随机分为3组:对照组、癫痫模型组和Res治疗组。采用重复低剂量氯化锂-联合皮罗卡品腹腔注射制备癫痫持续状态SD大鼠模型,并同时给予Res预防性治疗,对实验动物按痫性分级标准进行行为学观测,采用化学比色法检测痫性发作90min后模型动物大脑皮质过氧化氢酶(catalase,CAT)、谷胱甘肽(glutathion,GSH)、超氧化物歧化酶(superoxidedismutase,SOD)和谷胱甘肽还原酶(glutathion reductase,GR)含量变化。采用免疫组织化学法和免疫印迹法(Western blot)检测痫性发作90 min后模型动物大脑皮质中凋亡相关蛋白Bcl-2、Caspase-3和Bax的表达变化。结果:癫痫大鼠大脑皮质CAT、GR、SOD和GSH的含量较对照组显著减少(P〈0.05),凋亡相关蛋白Bcl-2表达下调,该蛋白阳性细胞数减少,而caspase-3和Bax表达显著上调(P〈0.05),两种蛋白阳性细胞数增多(P〈0.05);Res预防性治疗后,治疗组动物痫性发作潜伏期较模型组显著延长(P〈0.05);药物治疗明显上调模型动物大脑皮质中巯醇抗氧化剂(酶)CAT、GR、SOD和GSH的含量(P〈0.05),增加抗凋亡蛋白Bcl-2表达(P〈0.05)和阳性细胞数(P〈0.05),同时抑制促凋亡因子Caspase-3和Bax的表达(P〈0.05)和减少两种蛋白阳性细胞数(P〈0.05)。结论:Res能通过上调癫痫模型大鼠皮质内源性巯基抗氧化物(酶)CAT、GR、SOD和GSH的含量,调控凋亡相关蛋白Bcl-2、caspase-3和Bax的表达变化,对抗SE诱导的神经细胞氧化损伤,发挥神经保护作用。
Objective: This study investigated the antioxidative mechanism of the resveratrol’s in status epilepticus(SE) rats model induced by treating LiCl plus repeated low doses of pilocarpine.Methods: Fifty four SD-rats were divided into three groups randomly: sham group,model group and treatment group.SE model rats were established by injecting LiCl plus repeated low doses of pilocarpine into abdominal cavity.Meantime,rats were treated by peritoneally injecting Resveratrol(30 mg/kg).Then the thiol antioxidants including catalase(CAT),glutathione reductase(GR),superoxide dismutase(SOD)and glutathione(GSH) activities were examined by colorimetric method in different groups.The protein expression of the Bcl-2,caspase-3 and Bax were measured by Western blot and immunohistochemistry in three groups.Results: The SE model rats exhibited a significant decrease of the concentration of CAT,GR,SOD,GSH and Bcl-2,but an increase of the concentration of Bax and caspase-3 compared to sham group(P〈0.05).After treatment with Resveratrol,the treatment group rats exhibited a significant increase in seizure latency,the concentration of the CAT,GR,SOD,GSH,and Bcl-2(P〈0.05),but a decrease in the concentration of caspase-3 and Bax compared to model group(P〈0.05).Conclusion: The results from our study indicate that Resveratrol could resist the oxidative damage of central nervous systems in SE model rats by up-regulating the expression of the thiol antioxidants(including CAT,GR,SOD and GSH) and down-regulating the protein expression of the caspase-3 and Bax.