托吡酯对大鼠局灶性脑缺血再灌注的保护机制

Protective effect of topiramate on cerebral ischemia-reperfusion injury in rats

目的探讨托吡酯对大鼠局灶性脑缺血再灌注的保护机制。方法采用拴线法建立大鼠(购自上海斯莱克实验动物有限责任公司)大脑中动脉局灶缺血再灌注损伤模型,并将造模成功大鼠随机分为模型组、托吡酯组,另外设立假手术组,每组10只大鼠,并于术后24 h评价大鼠神经行为变化,测定大鼠脑梗死体积及脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNLE)阳性细胞数,测定肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6 mRNA。所有数据均用SPSS 17.0统计分析,采用t检验分析。结果与假手术组比较,模型组大鼠神经行为评分[(2.85±0.32)分比(0±0)分,t=4.764,P<0.01]、脑梗死体积[(114.68±11.47)比(0±0),t=4.375,P<0.01]、原位缺口末端标记法(TUNEL)阳性细胞数[(48.92±4.89)个比(5.54±0.55)个,t=4.903,P<0.01]、TNF-α[(2.13±0.21)比(1.00±0.13),t=4.678,P<0.01]、IL-1β[(1.89±0.13)比(1.00±0.08),t=4.532,P<0.01]及IL-6 mRNA表达量[(1.98±0.16)比(1.00±0.05),t=4.893,P<0.01],TNF-α[(28.38±2.84)比(12.57±1.26),t=4.679,P<0.01]、IL-1β[(83.89±8.13)比(25.79±2.58),t=4.421,P<0.01]及IL-6[(63.58±6.36)比(33.48±3.35),t=4.249,P<0.01]含量,磷酸化核因子κB抑制蛋白α(p-IκBα)[(0.88±0.07)比(0.07±0.01),t=5.890,P<0.01]及磷酸化核因子-κB(p-NF-κB)p65[(2.32±0.22)比(0.15±0.01),t=5.432,P<0.01]表达量均提高,差异有统计学意义;与模型组比较,托吡酯组大鼠神经行为评分[(1.46±0.15)分比(2.85±0.32)分,t=5.786,P<0.01]、脑梗死体积[(70.38±7.40)比(114.68±11.47),t=5.370,P<0.01]、TUNLE阳性细胞数[(10.89±1.10)个比(48.92±4.89)个,t=5.421,P<0.01]、TNF-α[(1.42±0.14)比(2.13±0.21),t=4.345,P<0.01]、IL-1β[(1.02±0.13)比(1.89±0.13),t=5.124,P<0.01]及IL-6 mRNA表达量[(1.08±0.12)比(1.98±0.16),t=4.875,P<0.01],TNF-α[(15.42±1.54)比(28.38±2.84),t=5.658,P<0.01]、IL-1β[(45.32±4.53)比(83.89±8.13),t=5.378,P<0.01]及IL-6含量[(44.38±4.38)比(63.58±6.36),t=4.209,P<0.01],p-IκBα[(0.18±0.02)比(0.88±0.07),t=5.432,P<0.01]及p-NF-κB p65表达量均降低,差异有统计学意义[(0.16±0.02)比(2.32±0.22),t=5.569,P<0.01]。结论托吡酯能显著的抑制缺血再灌注大鼠脑损伤,与阻断NF-κB信号通路有关。

Objective To explore protective effect of Topiramate on cerebral ischemia-reperfusion injury in rats.Methods The rat model of middle cerebral artery ischemia-reperfusion injury was established by tethering.The successful modeling rats were randomly divided into model group,topiramate group,sham operation group was estabished,10 rats in each group.Twenty four hours after the operation,neurological behavior score,cerebral infarction volume,terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay(TUNEL)positive cell number,the expression of tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6 mRNA and protein,and the expression of nuclear factor-κB(NF-κB)signal pathway related protein was detected.Results Compared with sham group,rats’neurological behavior score[(2.85±0.32)vs.(0±0),t=4.764,P<0.01]、cerebral infarction volume[(114.68±11.47)vs.(0±0),t=4.375,P<0.01],TUNEL positive cell number[(48.92±4.89)cells vs.(5.54±0.55)cells,t=4.903,P<0.01],the expression of TNF-α[(2.13±0.21)vs.(1.00±0.13),t=4.678,P<0.01],IL-1β[(1.89±0.13)vs.(1.00±0.08),t=4.532,P<0.01]and IL-6 mRNA[(1.98±0.16)vs.(1.00±0.05),t=4.893,P<0.01],the expression of TNF-α[(28.38±2.84)vs.(12.57±1.26),t=4.679,P<0.01],IL-1β[(83.89±8.13)vs.(25.79±2.58),t=4.421,P<0.01]and IL-6[(63.58±6.36)vs.(33.48±3.35),t=4.249,P<0.01]protein,the expression of phosphorylated inhibitor of NF-κB(p-IκBα)[(0.88±0.07)vs.(0.07±0.01),t=5.890,P<0.01] and p-NF-κB p65[(2.32±0.22)vs.(0.15±0.01),t=5.432,P<0.01]was increased in model group.Compared with model group,rats’neurological behavior score[(1.46±0.15)vs.(2.85±0.32),t=5.786,P<0.01],cerebral infarction volume[(70.38±7.40)vs.(114.68±11.47),t=5.370,P<0.01],TUNLE positive cell number[(10.89±1.10)cells vs.(48.92±4.89)cells,t=5.421,P<0.01],the expression of TNF-α[(1.42±0.14)vs.(2.13±0.21),t=4.345,P<0.010],IL-1β[(1.02±0.13)vs.(1.89±0.13),t=5.124,P<0.01]and IL-6 mRNA[(1.08±0.12)vs.(1.98±0.16),t=4.875,P<0.01],the expression of TNF-α[(15.42±1.54)vs.(28.38±2.84),t=5.658,P<0.01],IL-1β[(45.32±4.53)vs.(83.89±8.13),t=5.378,P<0.01]and IL-6 protein[(44.38±4.38)vs.(63.58±6.36),t=4.209,P<0.01],the expression of p-IκBα[(0.18±0.02)vs.(0.88±0.07),t=5.432,P<0.01]and p-NF-κB p65[(0.16±0.02)vs.(2.32±0.22),t=5.569,P<0.01]was decreased in Topiramate.Conclusion Topiramate could significantly inhibit brain injury induced by ischemia-reperfusion in rats,which related to blocking the signal pathway of NF-κB.