平痫颗粒对癫痫大鼠海马神经元细胞凋亡及基因表达的影响

Effects of Pingxian Particle on Cell Apoptosis of Hippocampal Neurons and Its Gene Expression in Epileptic Rat Model

目的探讨平痫颗粒在阻抗癫痫形成中对大鼠海马区神经元细胞凋亡影响的分子生物学机制。方法 60只45日龄Wistar大鼠随机分为正常对照组、模型组、阳性对照组及平痫小、大剂量组。除正常对照组外,其余各组大鼠均以戊四唑亚惊厥剂量35 mg/kg腹腔注射诱导癫痫模型。同时平痫大、小剂量组以1.66、0.42 g/mL原药材含量2 mL/d灌胃,阳性对照组给予等量的3.6 mg/mL苯巴比妥混悬液灌胃,模型组、正常对照组胃饲等量双蒸水,持续5周。HE染色观察海马神经元细胞病理形态学改变,免疫组化染色(SABC法)观测bcl-2、bax蛋白阳性细胞的表达。结果给大鼠连续注射戊四唑过程中,模型组大鼠1～4周逐渐出现Ⅰ～Ⅵ级癫痫发作。各治疗组发作次数和级别明显降低,最高为Ⅳ级,组间发作级别、次数相比差异无统计学意义。HE染色显示,各治疗组海马神经元细胞病理表现改善明显。第3、5周末模型组bcl-2表达明显低于正常对照组(P<0.01),bax表达明显高于正常对照组(P<0.01);各治疗组bcl-2表达均明显高于模型组(P<0.01),bax表达均明显低于模型组(P<0.01)。结论平痫颗粒可能是通过促进bcl-2蛋白并抑制bax蛋白的表达,从而减少神经元细胞的凋亡,以达到阻抗癫痫的目的。

Objective To investigate the molecular biological mechanism of Pingxian Particle＇s anti-epileptic effects on hippocampus neurons apoptosis. Methods Sixty Wistar rats were randomly assigned to normal control group, model group, positive control group, Pingxian large dose group and Pingxian small dose group. Except the normal control group, all the groups were intraperitoneally injected with 35 mg/kg pentylenetetrazol to induce epilepsy model. Pingxian large dose （1.66 g/mL） and small dose （0.42 g/mL） groups were intragastrically infused with Pingxian Particle 2 mL/d, the positive control group received 3.6 mg/mL phenobarbital suspension, the normal control group and model group were drenched with distilled water 2 mL/d for 5 weeks. The pathological changes of hippocampal neurons were observed dynamically by HE staining. The bcl-2 and bax protein positive cells were labeled with immunohistochemical SABC to dynamically observe the protein expression. Results The seizttres of model group became more and more serious with kindle times, and the rats appeared grade I-VI behavior during lst--4th week. The onset times and onset grade of treatment groups were fewer and lower, and the highest grade were only IV, there were no significant differences in the onset grade and times between every two groups. The pathology represents of hippocampal neurons of treatment groups improved obviously. At the end of 3rd and 5th week, the number of bcl-2 protein expression at hippocampus in the model group was obviously less than the normal control group {P〈0.01）, and that of Pingxian groups and positive control group were obviously more than the model group （P〈0.01）. The number of bax protein expression at hippocampus in the model group was obviously more than the normal control group （P 〈 0.01）, and that of Pingxian groups and positive control group were obviously less than the model group （P〈0.01）. Conclusion Pingxian Particle may treat epilepsy though promoting the expression of bcl-2 and restraining the expression of bax.