宁动颗粒对多巴胺能神经元代谢的影响

Effect of Ningdong Granules on metabolism of dopaminergic neurons

目的研究宁动颗粒对多巴胺能神经元细胞代谢的影响。方法将12只18 d胎龄胎鼠随机分为宁动颗粒组(NDG)、对照组(CTR)、氟哌啶醇组(Hal)、正常组(Normal),每组3只。将每组的多巴胺能神经细胞分别提取分离并培养,采用免疫荧光间接法对多巴胺能神经元细胞进行酪氨酸羟化酶(TH)抗染色;采用神经元轴突微管蛋白anti-Tau1结合FITC荧光(绿光)Ⅱ抗标记、神经元轴突微管蛋白anti-Tau1结合Cy 3荧光(红光)Ⅱ抗标记,分别对多巴胺能神经元细胞及神经元轴突进行荧光染色鉴定,采用酶联免疫吸附法(ELISA)检测18 d胎龄胎鼠DA能神经细胞在药物干预前及干预1、6、12、24 h后多巴胺能神经元细胞培养液中单胺氧化酶(MAO)、DA、5-羟色胺(5-HT)、高香草酸(HVA)、5-羟吲哚乙酸(5-HIAA)含量,并记录各组DA能神经元细胞的代谢情况。结果与同组0 h比较,NDG组干预6、12 h后细胞培养液中MAO、DA、5-HT、HVA含量显著升高(P <0.05);与CTR组同期比较,Hal组干预0～24 h后细胞培养液中MAO、DA、5-HT、HVA含量差异无统计学意义(P> 0.05);干预0～24 h后,各组胎鼠培养液中5-HIAA含量差异无统计学意义(P> 0.05)。结论宁动颗粒能促进多巴胺能神经元代谢,但对5-HT的代谢无显著性影响。

Objective To study the effect of Ningdong Granules on the metabolism of dopaminergic(DA)neurons.Methods Twelve 18-days-old fetuses were randomly divided into Ningdong Granules(NDG)group,control(CTR)group,haloperidol(Hal)group and normal(Normal)group,with 3 rats in each group.DA neurons of each group were separately isolated and cultured,and tyrosine hydroxylase(TH)anti-staining was performed on DA neuron cells by immunofluorescence indirect method.Neuron axon anti-Tau1 anti-labeling combination with FITC fluorescence(green light)Ⅱanti-labeling,neuron axon anti-Tau1 labeling combined with Cy 3 fluorescence(red light)Ⅱanti-labeling,DA neuron cells and neuron axons were identified by fluorescent staining.The contents of MAO,DA,5-HT,HVA and 5-HIAA in DA neuron cell culture medium of 18-days-old fetuses before and 1,6,12 and 24 h after drug intervention were detected by ELISA,and the metabolism of metacells of DA energetic spirit in each group was recorded.Results Compared with 0 h,the contents of MAO,DA,5-HT and HVA in the cell culture medium of NDG group increased significantly after 6 and 12 h of intervention(P<0.05);compared with CTR group at the same time,there was no significant difference in the contents of MAO,DA,5-HT and HVA in the cell culture medium of Hal group after 0-24 h of intervention(P>0.05);after 0-24 h of intervention,the 5-HIAA content in cell culture of fetuses in each group had no significant difference(P>0.05).Conclusion NDG can promote the metabolism of DA neurons,but has no significant effect on the metabolism of 5-HT.