托吡酯对蛛网膜下腔出血后神经细胞凋亡影响及机制的实验研究  

作　　者：曹杰[1] 范成普 杜杭根[2] 周冰之[2] 王承[2] 郝必烈[2] 

机构地区：[1]浙江中医药大学第二临床医学院,310053 [2]浙江中医药大学附属第二医院,310005

出　　处：《浙江临床医学》2017年第10期1783-1785,共3页Zhejiang Clinical Medical Journal

基　　金：浙江中医药大学校级科研基金资助项目（2015ZY20）;浙江省教育厅一般科研项目资助项目（Y201636786）

摘　　要：目的从神经细胞凋亡角度探讨托吡酯（TPM）对实验性大鼠蛛网膜下腔出血（SAH）后早期脑损伤（EBI）的神经保护作用，从而为临床应用提供理论依据。方法取140只健康雄性SD大鼠，随机分为5组，分别为假手术组（20只）、SAH模型组（30只）和SAH＋TPM干预组（90X平均分为3组，术后2h分别予TPM20mg＆g、40mg／kg及80mg／kg腹腔注射）。利用颈内动脉穿刺法制作蛛网膜下腔出血模型，对各组在制模后24h进行死亡率统计和神经行为学评分，采用干湿重法检测脑水肿指数，TUNEL／NeuN／DAPI／Merge荧光染色观察神经元细胞凋亡，Western blot法评估Caspase-3及Bcl-2、Bax凋亡相关蛋白表达情况。结果与假手术组比较，SAH模型组和各TPM干预组的死亡率和脑水肿指数上升，神经学评分降低。与SAH模型组比较，3个TPM干预组的神经学评分均上升，脑水肿指数均下降，差异有统计学意义（P〈0．05）。大鼠SAH后24h，Caspase-3YZBcl-2、Bax凋亡相关蛋白表达增强，凋亡细胞增加。TPM干预后，Caspase-3和促凋亡蛋白Bax表达减少，抑制凋亡蛋白Bcl-2表达增加，凋亡细胞数目有所减少，且这种效应与剂量呈正相关。结论TPM可能通过抑制Caspase。3、Bax表达，促进Bcl-2表达，从而减轻SAH后神经元细胞继发性凋亡损伤，在神经功能保护方面具有积极作用。Objective To explore the neuroprotective effect of topiramate on brain injury in rats after experimental subarachnoid hemorrhage from the perspective of neuronal apoptosis, so as to provide theoretical basis for clinical application. Methods Forty healthy male SD rats were randomly divided into 5 groups： sham operation group （ 20 rats ） , SAH mode/group （ 30 rats ） and SAH ＋ TPM intervention group （ 90 rats were divided into 3 groups, 2 hours after operation TPM 20mg/kg, 40mg/kg and 80mg/kg intraperitoneal injection ） . The model of mortality and neurobehavioral scores were made in each group at 24 h after model establishment. The brain edema index was measured by dry and wet weight method. TUNEL/NeuN/DAPI/Merge fluorescence staining were used to observe the apoptosis of neurons. The expression of Caspase-3, Bcl-2 and Bax-related proteins was evaluated by Western blot. Results Compared with the sham group, the mortality and brain edema index of the SAH model group and each TPM intervention group was increased and the neurological score was decreased. Compared with SAH model group, the neurological scores of the three TPM intervention groups was increased and the brain edema index was decreased, the differences were statistically significant （ P〈0.05 ） . After 24 hours of SAH, the expression of Caspase-3, Bcl-2 and Bax was up-regulated and the number of apoptotic cells were increased. After TPM intervention, the expression of Caspase-3 and pro-apoptotic protein Bax was decreased, the expression of Bcl-2 and the number of apoptotic cells were decreased, and the effect was positively correlated with the dose. Conclusions TPM may promote the expression of Bcl-2 by inhibiting the expression of Caspase-3 and Bax, thereby reducing the secondary apoptosis of neurons after SAH, and has a positive effect on neurological function protection.

关 键 词：托吡酯 蛛网膜下腔出血 早期脑损伤 细胞凋亡 凋亡相关蛋白 

分 类 号：R743.35[医药卫生—神经病学与精神病学]