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目的探讨反复惊厥对新生大鼠大脑皮层神经元迁移蛋白DCX、LIS1蛋白表达的影响。方法将48只生后7天的Sprague-Dawley大鼠随机分为对照组和惊厥组各24只,惊厥组每日吸入三氟乙醚诱导惊厥发作1次,每次持续30 min,连续6天;对照组不吸入三氟乙醚。两组大鼠分别于日龄14、17、21天各选取8只大鼠断头取脑,用Western Blot技术及免疫组织化学染色观察大鼠大脑皮层DCX、LIS1表达的变化。结果随着日龄增加,对照组新生大鼠大脑皮层DCX和LIS1免疫组化累计光密度(AOD)逐渐降低。与对照组相比,日龄14天时,惊厥组大鼠大脑皮层DCX和LIS1免疫组化AOD值明显降低[DCX:(2920±152)比(3718±222),LIS1:(8146±367)比(11 965±385),P<0.05],17天时大脑皮层LIS1免疫组化AOD值明显增高[(8881±333)比(7164±286),P<0.05],21天时大脑皮层DCX免疫组化AOD值明显增高[DCX:(2776±135)比(2379±110),P<0.05]。Western Blot示与对照组比较,日龄14天时,惊厥组大脑皮层DCX蛋白表达明显降低(P<0.05),21天时,惊厥组大脑皮层LIS1蛋白表达明显增高(P<0.05)。结论LIS1、DCX在正常新生大鼠大脑皮层中表达随时间进展表现出一定的规律性。反复惊厥影响新生大鼠大脑皮层神经移行相关蛋白DCX和LIS1蛋白表达,DCX和LIS1蛋白可能参与惊厥对脑发育的影响。
Objective To investigate the effects of recurrent seizures on the expression of DCX and LIS1 protein in cultured neonate rat cortical neurons. Methods Forty-eight Sprague-Dawley rats (7 days after birth) were randomly divided into control group and convulsion group (24 rats in each group). Convulsion group were infused with trifluoroethyl ether to induce convulsion seizure once every 30 minutes for 6 days. The control group did not inhaled trifluoroethyl ether. The rats in the two groups were decapitated on the 14, 17 and 21 days of age respectively, and the brain was cut out. The changes of DCX and LIS1 expression in the cerebral cortex were observed by Western Blot and immunohistochemical staining. Results With the increase of age, the cumulative optical density (AOD) of DCX and LIS1 in cerebral cortex of control group decreased gradually. Compared with the control group, the AOD values of DCX and LIS1 in the cerebral cortex of the convulsion group were significantly decreased at 14 days of age [DCX: (2920 ± 152) vs (3718 ± 222), LIS1: (8146 ± 367) (11 965 ± 385), P <0.05]. After 17 days, the AOD of LIS1 in the cerebral cortex was significantly higher than that in the control group [(8881 ± 333) vs (7164 ± 286), P <0.05] AOD was significantly higher [DCX: (2776 ± 135) vs (2379 ± 110), P <0.05]. Western Blot showed that compared with the control group, the expression of DCX protein in cerebral cortex of convulsion group was significantly decreased at 14 days of age (P <0.05). At 21 days, the expression of LIS1 protein in cerebral cortex of convulsion group was significantly increased (P <0.05). Conclusion The expression of LIS1 and DCX in normal neonate rat cerebral cortex shows some regularity with the progress of time. Repeated convulsions affect the expression of DCX and LIS1 protein in neonate rat cortical neurons, and the expression of DCX and LIS1 protein may be involved in the effects of convulsion on brain development.