茸菖胶囊对癫痫后认知障碍幼年大鼠神经发生的影响

Effect of Rongchang Capsule on the Neurogenesis in Young Rats with Cognitive Impairment after Epilepsy

目的观察茸菖胶囊对癫痫后认知障碍幼年大鼠学习记忆能力和神经发生的影响,探讨其治疗癫痫后认知障碍的可能机制。方法 102只SD大鼠随机分为正常组、模型组、中药高剂量组、中药中剂量组、中药低剂量组、西药组。运用戊四唑点燃法复制癫痫大鼠模型,并经水迷宫实验筛选认知障碍大鼠。正常组不予处理,模型组予生理盐水灌胃,中药高、中、低剂量组分别予浓度为0.44g/ml、0.22g/ml、0.11g/ml茸菖胶囊溶液灌胃,西药组予40mg/ml德巴金溶液灌胃,每次3ml,每日2次,均给药28d。每组又分别于5-溴脱氧尿苷嘧啶(BrdU)标记后24h和28d取材。采用水迷宫实验于治疗前后观察大鼠逃避潜伏期和跨越平台次数;观察BrdU阳性细胞形态并计数进行BrdU免疫组化评分,计算新生神经细胞迁移比率(Br-dU/DCX)、新生神经细胞分化为神经元比率(BrdU/NeuN)和新生神经细胞分化为胶质细胞比率(BrdU/GFAP)。结果治疗后西药组和中药高、中剂量组大鼠逃避潜伏期较模型组缩短(P<0.05或P<0.01),并且中药高剂量组较中药中、低剂量组大鼠逃避潜伏期缩短(P<0.05)。中药高剂量组大鼠跨越平台的次数显著高于模型组和中药低剂量组(P<0.05)。与模型组比较,24h时中药高剂量组、西药组和28d时中药高剂量组、中药中剂量组、西药组BrdU免疫组化评分显著减少(P<0.05或P<0.01)。西药组、中药高剂量组、中药中剂量组BrdU/DCX、BrdU/NeuN低于模型组(P<0.05或P<0.01);中药高剂量组BrdU/NeuN明显低于中药中、低剂量组(P<0.01);各组大鼠BrdU/GFAP比较差异无统计学意义(P>0.05)。结论茸菖胶囊可改善癫痫后认知障碍大鼠学习记忆功能,其机制可能为抑制大鼠海马齿状回神经前体细胞增、异常迁移和分化。

Objective To observe the effect and mechanism of Rongchang Capsule [capsule of Lurong （Cornu Cervi Panto trichum） and Shichangpu （Rhizoma Acori Tatarinowii）] on the neurogenesis and learning and memory abilities in young rats with cognitive impairment after epilepsy. Methods The Sprague-Dawley rats were randomized into the normal group （n= 16）, model group （n=18）, high-dose Chinese medicine group （n： 16）, middle-dose Chinese medicine group （n= 18）, low-dose Chinese medicine group （n： 16） and western medicine group （n： 18）. The rat models with cognitive impairment after epilepsy were induced by pentylenetetrazol kindling. The normal group was not treated. The model group was intragastricaliy adminis- tered with normal saline. The high-dose, middle-dose and low-dose Chinese medicine groups were intragastrically administered with 0.44 g/ml, 0.22 g/ml and 0.11 g/ml Rongchang Capsule solution. The western medicine group was intragastrically ad- ministered with 40 mg/ml Depakine Solution. Each group was given intragastric administration 3 ml once, twice daily for 28 days. The brain tissue samples were drawn 24 hours or 28 days after labeling with 5-bromodeoxyuridine pyrimidine （BrdU）. The escape latency and platform crossing times were observed with water maze.test before and after treatment. The morphology and number of BrdU-positive cell were detected and scored with BrdU immunohistoehemistry. The ratios of BrdU/doublecortin （DCX）, BrdU/neuron-specific nuclear protein （NeuN） and BrdU/glial fibrillary acidic protein （GFAP） were calculated. Results Comparing with the model group, the escape latency in the western medicine group, high-close Chinese medicine group and middle-dose Chinese medicine group was significantly shortened （P^0.05 or P^0.01）, and the shortening in the high- dose Chinese medicine group was more significant than those in middle-dose and low dose Chinese medicine groups （P〈0.05）. The platform crossing times in the high-dose Chinese medicine group were significantly higher than those in the model group and low-dose Chinese medicine group （P〈0.05）. Comparing with the model group, the BrdU immunohistochemistry scores were significantly decreased in the high-dose Chinese medicine group and western medicine group 24 hours after labeling withBrdU and in the high-dose Chinese medicine group, middle-dose Chinese medicine group and western medicine group 28 days af＇~er labeling with BrdU （P^0.05 or P-~0.01）. Comparing with the model group, the ratios of BrdU/DCX and BrdU/NeuN were significantly lower in the western medicine group, high-dose Chinese medicine group and middle-dose Chinese medicine group （P（0.05 or P（0.01）. The ratio of BrdU/NeuN in the high-dose Chinese medicine group was significantly lower than that in the middle-dose and low-dose Chinese medicine groups （P^0.01）. There was no significant difference between groups in the ratio of BrdU/GFAP （P^0.05）. Conclusion Rongchang Capsule can improve the learning and memory abilities in rats with cognitive impairment after epilepsy. Its mechanism may be relevant to inhibiting the proliferation, abnormal migration and differentiation of neural precursor cells in dentate gyrus.