熄风胶囊干预治疗对氯化锂-匹罗卡品癫痫大鼠海马突触损伤的影响

Effect of Xifeng capsule on hippocampal synaptic injury in epileptic rat induced by lithium chloride-pilocarpine

关键词:  癫痫  熄风胶囊  突触索  生长相关蛋白-43

Key words:  epilepsy  Xifeng capsule  synaptophysin  growth associated protein-43

摘要:

[目的] 观察中药熄风胶囊单药和联合用药干预治疗对氯化锂-匹罗卡品癫痫大鼠海马突触损伤的影响。[方法] 将SPF级健康雄性Wistar大鼠64只随机分为正常组、模型组、熄风胶囊高剂量干预组（熄高组）、熄风胶囊中剂量干预组（熄中组）、熄风胶囊低剂量干预组（熄低组）、熄风胶囊大剂量+托吡酯联合干预组（联高组）、熄风胶囊大剂量+托吡酯1/2剂量联合干预组（联低组）、托吡酯干预组（TPM组）各8只。运用氯化锂-匹罗卡品化学点燃法，复制癫痫大鼠模型，通过采用SABC免疫组化检测突触索（P38）表达水平来反映突触索的生长情况、原位杂交检测生长相关蛋白-43（GAP-43）mRNA的表达水平来反映GAP-43的水平。[结果] 1）所有癫痫大鼠海马CA1、CA3及齿状回分子层及颗粒细胞层均可见深染的颗粒状免疫反应产物，呈点片状分布。所有治疗组海马CA1、CA3起始层及齿状回内分子层P38免疫反应产物总面积显着低于模型组（P<0.01）.联高组海马CA1、CA3、齿状回区P38免疫反应产物总面积与正常组无统计学差异（P>0.05），熄高组海马CA1及CA3区阳性反应物与正常对照组无统计学差异（P>0.05）.在海马CA1区联高组和熄高组优于其余各治疗组（P<0.05）.在海马CA3区联高组作用优于熄低组和TPM组（P<0.05），与其余各治疗组比较无统计学差异（P>0.05）.在海马齿状回区联高组作用与其余各治疗组均有统计学差异（P<0.05）.2）各治疗组大鼠海马颗粒细胞GAP-43 mRNA表达显着低于模型组（P<0.01）.其中海马CA1区联高组和熄高组两组无统计学差异（P>0.05），优于熄中组和熄低组（P<0.05）.在海马CA3区，各治疗组之间无统计学差异（P>0.05）.在海马齿状回区联高组和熄高组与熄中组，熄低组和TPM组之间有统计学差异（P<0.05）.[结论] 熄风胶囊单药和联合用药能有效的干预氯化锂-匹罗卡品癫痫大鼠海马颗粒细胞层及内分子层P38和GAP-43的表达，从而起到减轻海马损伤的作用。

Abstract:

[Objective] To observe the intervention effect of Xifeng capsule only and combined therapy on hippocampal synaptic injury in epileptic rats induced with lithium chloride- pilocarpine. [Methods] The 64 SPF healthy male Wistar rats were randomly divided into normal group, model group, high dose of Xifeng capsule group (high group), medium dose of Xifeng capsule group (medium group), low dose of Xifeng capsule group (low group), high dose of Xifeng capsule+topiramate group (combined high group), high dose of Xifeng capsule + 1/2 dose of topiramate group (combined low group), topiramate group (TPM group). Every group had 8 rats. Applying the ignited method with lithium chloride-pilocarpine chemicals a model of epilepsy was reproduced in rats. Using SABC immunohistochemistry the synapse cable (P38) expression levels was detected to reflect the growth of synapses cable. The expression level of growth of associated protein-43 (GAP-43) mRNA was detected bu in situ hybridization to reflect the level of growth associated protein-43. [Results] 1)The experimental results showed that some visible point patchy distributed as stained granular immune reaction products was found in hippocampal CA1, CA3 and dentate gyrus molecular layer and the granule cell layer in all epileptic rats. The total area of P38 immune reaction products in hippocampal CA1, CA3 and dentate gyrus starting layer molecular layer of all treatment groups was significantly lower than that of the model group (P<0.01). The total area of P38 immune reaction products in hippocampal CA1, CA3 and dentate gyrus starting layer molecular layer of combined high group had no significant difference when compared with that of the normal group (P>0.05). Positive reaction products in hippocampal CA1 and CA3 areas of high group had no significant difference compared with the normal group also (P>0.05). In the hippocampal CA1 region combined high group and high group it was better than the low group and the rest of the treatment groups (P<0.05). In the hippocampal CA3 combined high group it was superior to the low group and TPM group (P<0.05), but showed no significant difference when compared with the rest of the treatment groups (P>0.05). In the dentate gyrus region, combined high group was significantly different compared with the rest of the treatment groups (P<0.05). 2)Hippocampal granule cells GAP-43 mRNA of all treatment groups was significantly lower than that of the model group (P<0.01). Combined high group had no significant difference with high group in hippocampal CA1 region (P>0.05), but was better than medium group and low group (P<0.05). In the CA3 region, there were no significant differences among all treatment groups (P>0.05). In the dentate gyrus region, combined high group and high group were significantly different compared with medium group, low group and TPM group (P<0.05). [Conclusion] Xifeng capsule only and combined therapy can effectively intervene the expression of growth-associated protein-43 in hippocampal granule cell layer and inner molecular layer synapses It plays a role in rats with epilepsy induced by lithium chloride-pilocarpine for reducing the hippocampal damage.