复方灯盏花滴丸对海马神经元一氧化氮和一氧化氮合酶的影响

目的研究复方灯盏花滴丸(FDD)对谷氨酸致大鼠原代海马神经元的NO和一氧化氮合酶的影响。方法取SD大鼠32只,每8只分别用生理盐水、尼莫地平、FDD高、低剂量灌胃,85h后,取每只大鼠股动脉血,制成5%的含药血清。另取新生24h SD乳鼠大脑原代培养海马神经元并鉴定后,随机分为对照组、尼莫地平组、FDD高、低剂量组和模型组,前4组分别加入上述配制对应的药物血清,模型组不加含药血清。以谷氨酸制成损伤模型。采用MTT法测定细胞存活率,酶法检测细胞释放NO量,细胞内总一氧化氮合酶(tNOS)和诱导型一氧化氮合酶(iNOS)活性。结果与对照组比较,模型组细胞存活率下降,NO、tNOS和iNOS活性增加(P0.01),与模型组比较,FDD高、低剂量组均能明显增加细胞存活率、减少NO释放、降低tNOS活性和iNOS活性(P0.05,P0.01)。结论 FDD对谷氨酸致原代培养大鼠海马神经元损伤有保护作用,其作用机制可能与降低一氧化氮合酶的活性、尤其iNOS活性及减少NO释放有关。

The effcts of Fu Fang Deng Zan Hua Drop Pill on nitric oxide and nitric oxide synthase in hippocampal neurons

Objective To explore the effect of Fu Fang Deng Zan Hua Di Wan(FDD) on glutamate (Glu) excitotoxicity in cultured primary hippocampal neurons of rats and to provide an experimental basis for FDD to be applied to the treatment of patients with cerebal ischemia injury. Methods Seropharmacological method was used to obtain the drug-containing serum.The primary hippocampal neurons of rat were cultured for 10 days and divided into 5 groups:control,model, nimodipine,5.00 g/kg FDD,and 1.25 g/kg FDD.The injury model was established by exposing hippocampal neurons to 500 μmol/L Glu for 20 minutes,5% drug-containing serum was added to all groups of cultured neurons except the model group.The following indexes were observed: the viability of cultured hippocampal neurons by MTT assay,NO content,the activity of total nitric oxide synthase(tNOS) and induced nitric oxide synthase(iNOS).Results Compared with control group,the viability of cultured hippocampal neurons in model group decreased,NO content,the activity of tNOS and iNOS were increased obviously,but when the cells were pretreated with the serum containing FDD,the above Glu-induced changes were prevented,the number of surviving cells increased significantly,NO content,the activity of tNOS and iNOS decreased markedly compared with model group.Conclusion FDD has protective effects against Glu excitotoxicity to cultured primary hippocampal neurons of rats,which possibly related with reducing NO content,the activity of tNOS and iNOS.