神经生长因子过表达治疗帕金森病模型大鼠的效果及其分子机制  

Efficacy and Molecular Mechanism of Nerve Growth Factor Overexpression in the Treatment of Parkinson’s Disease Model Rats

作　　者：燕燕[1] 崔传举[1] 王玉香[1] 李艾帆[1] YAN Yan;CUI Chuanju;WANG Yuxiang;LI Aifan(Department of Neurology, Zhengzhou First People’s Hospital, Zhengzhou Henan 450000, China)

机构地区：[1]郑州市第一人民医院神经内科,郑州450000

出　　处：《河南医学高等专科学校学报》2022年第1期1-5,共5页Journal of Henan Medical College

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ201909083)。

摘　　要：目的探讨神经生长因子(nerve growth factor,NGF)过表达诱导的脐血干细胞(umbilical cord mesenchymal stem cells,UMSCs)向多巴胺能神经元分化治疗帕金森病(Parkinson’s disease,PD)模型大鼠的效果及其对Tribbles同源蛋白3(TRB3)介导的信号通路的影响。方法建立PD模型,体外培养及诱导UMSCs,将pcDNA-NC、pcDNA-NGF转染至UMSCs中,分为pcDNA-NC组和pcDNA-NGF组。实验设置4组:NC组(大鼠仅进行手术,不进行注射)、PD组(PD模型)、UMSCs组(正常大鼠中注射含有pcDNA-NGF的UMSCs细胞悬液)、UMSCs+PD组(PD模型大鼠中注射含有pcDNA-NGF的UMSCs细胞悬液),每组各9只。采用实时荧光定量聚合酶链反应(qRT-PCR)检测NGF、神经胶质纤维酸性蛋白(GFAP)、神经元细胞内微管相关蛋白2(MAP2)、微管蛋白(Tubulin)、TRB3 mRNA的表达量;观察各组大鼠旋转圈数;采用免疫组化法检测各组大鼠黑质区酪氨酸羟化酶(TH)阳性细胞数;蛋白免疫印迹法(Western blot)检测NGF、TRB3的蛋白表达量。结果与pcDNA-NC组比较,pcDNA-NGF组NGF mRNA及蛋白、巢蛋白(Nestin)、GFAP、MAP2、Tubulin mRNA的表达水平升高(P<0.05)。与NC组比较,PD组大鼠旋转圈数增多(P<0.05);与UMSCs组比较,UMSCs+PD组大鼠旋转圈数增多(P<0.05);与PD组比较,UMSCs+PD组大鼠旋转圈数减少(P<0.05)。与NC组比较,PD组大鼠黑质区TH阳性细胞计数减少(P<0.05);与PD组比较,UMSCs+PD组大鼠黑质区TH阳性细胞计数增多(P<0.05)。与NC组比较,PD组大鼠脑组织中TRB3 mRNA及蛋白表达水平升高(P<0.05);与PD组比较,UMSCs+PD组大鼠脑组织中TRB3 mRNA及蛋白表达水平降低(P<0.05)。结论NGF过表达诱导的UMSCs可促进细胞向多巴胺能神经元分化,从而改善PD大鼠的症状,其作用机制可能是通过调控TRB3信号通路而发挥作用。Objective To investigate the effect of umbilical cord mesenchymal stem cells(UMSCs)induced by nerve growth factor(NGF)overexpression into dopaminergic neurons on Parkinson’s disease(PD)model rats and its effect on TRB3-mediated signaling pathway.Methods A PD model was established,and UMSCs were cultured and induced in vitro and pcDNA-NC and pcDNA-NGF were transfected into UMSCs,which were divided into pcDNA-NC group and pcDNA-NGF group.The experimental was set up into 4 groups:NC group(rats only operated without injection),PD group(PD model),UMSCs group(normal rats were injected with UMSCs cell suspension containing pcDNA-NGF),UMSCs+PD group(PD model rats were injected with a suspension of UMSCs cells suspension containing pcDNA-NGF),9 rats in each group.qRT-PCR was used to detect the expression of NGF,GFAP,MAP2,Tubulin,and TRB3 mRNA.The number of rotations of each group of rats were observed.The number of TH positive cells in the substantia nigra region of each group was detected by immunohistochemistry.Western blot was used to detect the protein expression of NGF and TRB3.Results Compared with the pcDNA-NC group,the expression levels of NGF mRNA and protein,Nestin,GFAP,MAP2,and Tubulin mRNA in the pcDNA-NGF group were significantly increased(P<0.05).Compared with the NC group,the number of rotations in the PD group was significantly increased(P<0.05).Compared with the UMSCs group,the number of rotations in the UMSCs+PD group was significantly increased(P<0.05).Compared with the PD group,the number of rotations in the UMSCs+PD group was significantly reduced(P<0.05).Compared with the NC group,the count of TH positive cells in the substantia nigra area of rats was significantly reduced(P<0.05).Compared with the PD group,the number of TH positive cells in the substantia nigra area in the UMSCs+PD group was increased significantly(P<0.05).Compared with the NC group,the expression levels of TRB3 mRNA and protein in the brain tissue of the PD group were significantly increased(P<0.05).Compared with PD group,the exp

关 键 词：帕金森病 神经生长因子 脐血干细胞 多巴胺能神经元 Tribbles同源蛋白3 

分 类 号：R-332[医药卫生]