熄风胶囊对氯化锂-匹罗卡品癫痫大鼠反复自发性发作及海马损伤的干预作用

The Intervention of Xifeng capsule on spontaneous recurrent seizures and hippocampalformation lesion in epileptic rats induced by Lithium-Pilocarpine

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-匹罗卡品癫痫大鼠反复自发性发作及海马损伤的影响。方法将SPF级健康雄性Wistar大鼠160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20只。运用氯化锂-匹罗卡品化学点燃法,复制癫痫大鼠模型,观察熄风胶囊单药和联合用药干预治疗对大鼠反复自发性发作(SRS)的影响;通过常规病理及电镜观察海马结构形态学改变,应用Timm组织化学染色法进行海马苔藓纤维出芽(MFS)的观察。结果 (1)所有干预组大鼠痫样发作平均级别均低于模型组而高于正常组,差异均有统计学意义(P<0.05)。(2)各干预组诱发癫痫持续状态的大鼠SRS发作次数均高于正常组而低于模型组,差异均有统计学意义(P<0.01),其中熄高组SRS次数低于熄中组、TPM组、熄低组,差异均有统计学意义(P<0.05);联髙组SRS次数低于熄低组,差异有统计学意义(P<0.01)。(3)Timm染色结果显示:所有干预组海马CA3始层及齿状回分子层Timm染色颗粒MFS总面积低于模型组,差异均有统计学意义(P<0.05),其中在CA3区熄高组、联髙组、联低组与模型组相比,差异有统计学意义(P<0.01)。(4)光、电镜结果显示:各干预组中联低组海马结构损伤最轻,熄高组海马结构损伤较熄中组、熄低组轻。结论熄风胶囊单药及与TPM联合用药可以有效地减低氯化锂-匹罗卡品癫痫大鼠的SRS,减轻海马损伤,抑制MFS。

Objective To study the intervention of Xifeng capsule single and combination effect on spontaneous recur- rent seizures（SRS） and hippocampal formation lesion in epileptic rats induced by Lithium-Pilocarpine. Methods All experi- mental rats were randomly divided into eight groups： normal control group, model group, Xifeng high-dose group, Xifeng mid- die-dose group, Xifeng low-dose group, Xifeng high-dose ＋ TPM group, Xifeng high-dose ＋ TPM 1/2 dose group, TPM group. We use lithium-pilocarpine chemical ignition method to duplicate the epileptic rat model. And with conventional his- topathological method and transmission electron microscope were utilized to observe morphological changes in hippocampal for- mation,Timm histochemical technique was adopted to study mossy fiber sprouting. Results （1）To compare the situation from the seizures in rats, the average attack level of all rats of Intervention group lower than model group（P〈0. 05）, but higher than the control group （P〈0. 01）. （2） All intervention group, the SRS frequencies obviously lower than model group（P〈 0.05）. （3）In all intervention group, the total aera of mossy fiber sprouting（MFS） of the hippocampal CA3 subfield and dentate gyrus were more than that of normal group（P〈0.05）. Both in the hippocampal CA3 subfield and dentate gyrus, the total aera of mossy fiber sprouting（MFS） have significance difference between Xifeng high-dose group and Xifeng low-dose group（P〈0.05）. （4）Using conventional histopathological method and transmission electron microscope, ultrastructure of neurons in hip-pocampus was regular in normal group but damaged obviously in model group. In all therapy groups. The damage of neuron in hippocampus was ameliorated in Xifeng high-dose ＋ TPM1/2 dose group, in Xifeng high-dose group. The damage of neuron in hippocampus was less than in Xifeng middle-dose group and in Xifeng low-dose group. Conclusion The intervention of Xifeng capsule single and combination can effectively reduce the degree of epileptic seizures in rats,protect hippocampal neurons from being more seriously and deterred hippocampal mossy fiber sprouting（MFS）.