丙戊酸单药治疗对癫癎患儿中性粒细胞功能影响的自身对照研究

Self control study of the effect of valproate acid monotherapy on neutrophilic function in epileptic children

目的探讨丙戊酸(VPA)单药治疗对癫癎患儿中性粒细胞功能的影响,进一步指导VPA的临床应用。方法选取复旦大学附属儿科医院2006年10月至2008年7月门诊首次确诊并给予VPA单药治疗的癫癎患儿。以二氢若丹明荧光染色外周血中性粒细胞,通过流式细胞仪分别检测患儿服药前和服药6个月时的中性粒细胞自身活化率和刺激指数;收集健康儿童全血,分别加适量VPA配成0、50、100、150和200μg.mL-1浓度进行培养,于培养后6、18和24h检测中性粒细胞自身活化率和刺激指数(SI),观察中性粒细胞功能变化的时间依赖性和浓度依赖性特征。结果研究期间纳入癫癎患儿29例,其中男13例,女16例;全身性发作22例,部分性发作7例;治疗前中性粒细胞自身活化率为(6.41±3.14)%,治疗后为(12.30±7.73)%;治疗前SI为440.6±32.4,治疗后为429.6±26.5;差异均有统计学意义。全身性发作和部分性发作间中性粒细胞自身活化率和SI差异均无统计学意义。中性粒细胞自身活化率随VPA浓度升高而增高,SI随培养时间的延长而显著降低,VPA 50μg·mL-1培养6h后中性粒细胞SI轻度升高,但培养24h后显著降低;VPA 200μg·mL-1培养后各时间点均见中性粒细胞SI显著降低。结论VPA单药治疗癫癎患儿体内存在中性粒细胞的自身活化现象,中性粒细胞自身活化率的升高呈浓度依赖性,SI的降低呈时间依赖性和剂量依赖性,VPA单药治疗可导致中性粒细胞功能的显著下降,且该作用与癫癎发作类型无关。

Objective To explore the effect of valproate acid （VPA） treatment on the neutrophilic function in epileptic children and analyze its mechanism,then further instruct clinic application of VPA. Methods Newly diagnosed epileptic children were recruited in the out-patient clinic of our hospital from October 2006 to July 2008. The auto-activation levels of neutrophils , including auto-activation rates and stimulation index, were detected before and after 6 months VPA treatment respectively by flow cytometry with dihydrorhodamine as fluorochrome. In vitro, the changes of neutrophilic function were explored at different VPA concentration （ control, 50 , 100 and 200 μg · mL^-1 respectively） and different time points （ 6,18 and 24 h respectively） through the culture of whole blood collecting from healthy children , and the feature of the changes of neutrophilic function was studied at different VPA concentrations and different time points. Results 29 newly diagnosed epileptic children were recruited, within which there were 13 boys and 16 girls, 22 general seizures and 7 focal seizures; their age ranged from 8-month-old to 193-monthold and the average age was 60.5 ±49 months before VPA treatment compared to 14-month-old to 200- month -old and 65.8 ±54.5 months after VPA treatment respectively. The auto-activation rate of neutrophils before VPA treatment was （6.41 ± 3.14）% ,while the rate was （ 12.30 ± 7.73 ） % after VPA treatment. The stimulation index was 440.6 ± 32.4 and 429.6 ± 26.5 , respectively. There were significant differences between the two groups in auto-activation rate of neutrophils and stimulation index（ P 〈 0.05 ）, but no difference between general seizures and focal seizures. In vitro, the neutrophlic function degraded significantly in a time-depending way with the culture time prolonged; at 50μg · mL^-1 VPA concentration, the neutrophilic function increased little after 6 h whole blood culture, but degraded strikely after 24 h whole blood culture ; at 200 μg · mL^-1 VPA concentration, the neutrophilic function dropped dramatically at all time points. Conclusions The phenomenon of auto-activation of neutrophils exists in epileptic children treated with VPA, the neutrophilic function degrades in a time-depending and concentration-depending way. VPA monotherapy ,though within the therapeutic blood concentration, will attenuate the neutrophlic function significantly. The impact of VPA on neutrophilc function has no relationship with the type of seizures. Maybe VPA can modulate neutrophihc function by influencing its phagocytosis and oxidation. Epileptic children＇s anti-infective function may be attenuated.