丹参对急性痫性痉挛大鼠模型的影响

The Influence of Salvia Militiorrhiza on Acute Seizure Model Rat

目的观察丹参对急性痫性痉挛模型大鼠行为学、脑电图和脑内胶质原纤维酸性蛋白(GFAP)表达的影响,观察丹参的抗痫效果。方法将24只大鼠随机分为戊四氮(PTZ)致痫组、丹参干预组和正常对照组,戊四氮致痫组大鼠腹腔注射戊四氮(75 mg/kg);丹参干预组先给大鼠腹腔注射丹参注射液(10 ml/kg),30 min后再腹腔注射戊四氮;正常对照组大鼠腹腔注射等量的生理盐水。大鼠注射戊四氮后,观察其行为学改变,描记脑电图。各组大鼠于戊四氮注射24 h和72 h后处死,取含有海马的脑组织行GFAP免疫组织化学染色,光镜观察、照相,并用图像分析系统对各组大鼠海马结构内GFAP阳性细胞进行计数和平均灰度值测定。结果 (1)行为学改变:丹参干预组大鼠痫性发作等级和发作时间明显小于戊四氮致痫组(P<0.05)。(2)脑电图显示,两组在发作期无明显的脑电图改变;在发作后恢复期,丹参组与PTZ组相比,脑电图出现棘波的频率低(1次/5s vs 8次/5s),波幅低。(3)免疫组织化学结果显示:①24 h组:与正常组大鼠相比,PTZ组海马内GFAP表达量增多,以齿状回和CA3区最为明显。GFAP阳性细胞突起增多、增粗、分支增多,细胞着色加深。丹参治疗组GFAP表达较注射PTZ组增多,阳性细胞数量增加,突起数目增多,长度增长,细胞着色更深。②72 h组:PTZ组和丹参组与正常组相比,GFAP的表达无明显差异。结论丹参可以明显降低急性痫性痉挛模型大鼠的发作等级和发作时间,癫痫发生后24 h海马内GFAP表达增多对机体可能有保护作用;丹参可能是通过促进GFAP表达增高来发挥治疗作用。

Objective In order to investigate the effect of Salvia militiorrhiza（SMR） on the acute seizure model,the behavior,electroencephalogram of the model and the expression of GFAP were observed within rat brain during the episode of epilepsy and after treatment.Methods 24 rats were divided into three groups randomly： PTZ（pentylenetetrazol） induced seizure group（10 rats）,SMR treatment group（10 rats） and normal control group（4 rats）.PTZ induced seizure group＇s rats were received a dose of 75 mg/kg/body weight PTZ intraperitoneally;Parenteral solution of SMR（10ml/kg） were injetcted into the SMR treatment group＇s rats abdominal cavity just 30 minutes prior to the injection of PTZ.Normal control group＇s rats were received same volume saline instead of PTZ.Behavior observation and EEG were observed through the behavior of the rats after injection of PTZ.Immunohistochemistry was performed on the GFAP-positive cells within hippocampus.Results（1） Behavior changes： The stages of seizure and the duration of convulsion of the SMR group were lower than that of PTZ group（P〈0.05）.（2） EEG records contained obvious epileptic electric discharge in PTZ group including abnormal spikes,polyspike discharge,and spike-wave complexes.The EEG records of SMR group contained epileptic electric discharge in a low degree,it showed intervallic polyspike discharge.（3） Immunohistochemistry results was shown that： ①24 h group： Compare with normal control group,the expression of GFAP within hippocampus of PTZ group was obviously increased,especially seen in dentate gyrus and CA3 region,wherein GFAP-positive cells had more branches,along with dense staining compare with PTZ group;the expression of GFAP in SMR treatment group was increased,wherein GFAP-positive cells＇ number and length were both increased.The staining of them was more condense than PTZ group.② 72h group： The expression of GFAP has no significant difference among PTZ group,SMR group and normal control group.Conclusion The inhibition of SMR on acute epileptic spasm is obvious;The increased expression of GFAP within hippocampus 24 hours after seizure perhaps has protection for animals,which suggests the GFAP perhaps takes part in SMR therapeutical effect.