摘要
目的探究芍药苷(PF)在神经干细胞(NSCs)移植治疗帕金森病(PD)过程中的作用。方法2024年2月至2024年9月进行细胞的提取、培养及分化。NSCs从孕13~16 d SD大鼠胚胎中分离培养1周,细胞计数试剂盒-8(CCK-8)检测不同浓度PF(0~500μmol/L)对NSCs活性的影响。设置对照组、PF组(100μmol/L)、XAV939组(2μmol/L)和PF+XAV939组(100μmol/L PF+2μmol/L XAV939)诱导分化,细胞免疫荧光评估分化效果。通过立体定向移植NSCs或NSCs+PF至纹状体,设置PD组(6-OHDA)、Sham组(假手术组)、NSCs组(2×106/10μl)、NSCs+PF组(2×106 NSCs+100μmol/L PF)评估移植疗效。阿扑吗啡诱导旋转试验评估模型稳定性,免疫组化及荧光染色分析移植细胞的存活及分化数量。组间比较采用t检验或方差分析。结果NSCs聚集成神经球(Nestin+)具有多向分化潜能(GFAP+、CNPase+、MAP2+)。100μmol/L PF的NSCs活性显著高于对照组(1.090±0.010比1.006±0.005,t=12.500,P<0.050),可诱导其向MAP2+成熟神经元分化,且PF激活WNT/β-catenin通路诱导TH+多巴胺能神经元分化,XAV939可阻断此效应。PD模型中TH+神经元低于假手术组(0.051±0.018比1.000±0.350,t=4.705,P<0.010)。移植PF+NSCs至PD大鼠纹状体后,旋转次数低于NSCs组(4.33±0.58比6.33±0.58,t=4.240,P<0.05),且PF+NSCs组TH+神经元高于NSCs组(0.699±0.105比0.151±0.101,t=6.532,P<0.05)。结论PF通过Wnt-β连环蛋白通路促进NSCs向多巴胺能神经元分化并增强其活性,改善PD大鼠运动功能。
Objective Parkinson’s disease(PD)is characterized by progressive loss of dopaminergic neurons,and neural stem cell(NSC)therapy has therapeutic potential.This study aimed to explore the role of paeoniflorin(PF)in the treatment of PD by NSC transplantation.Methods Cell extraction,culture and differentiation will be carried out from February 2024 to September 2024.NSCs were isolated from SD rat embryos at 13-16 days of gestation and cultured for a week.The effects of PF(0-500μmol/L)at different concentrations on the activity of NSCs were detected by cell counting kit(CCK-8).The differentiation of control group(0μmol/L PF),PF group(100μmol/L),XAV939 group(2μmol/L)and PF+XAV939 group(100μmol/LPF+2μmol/L XAV939)was induced,and cell immunofluorescence was used to evaluate the differentiation efficiency.NSCs or NSCs+PF were stereotaxically transplanted into the striata.PD group(6-OHDA),Sham group(sham operation),NSCs group(2×106/10μl),and NSCs+PF group(2×106 NSCs+100μmol/L PF)were set up to evaluate the transplantation efficacy.The model stability was evaluated by apomorphine induced rotation test,and the survival and differentiation of transplanted cells were analyzed by immunohistochemistry and fluorescence staining.Comparison between groups was performed by t test or ANOVA.Results NSCs aggregate into neurospheres(Nestin+)and have multi-directional differentiation potential(GFAP+,CNPase+,MAP2+).The activity of NSCs in 100μmol/L PF was significantly higher than that in the control group(1.090±0.010 vs.1.006±0.005,t=12.500,P<0.05),which could induce their differentiation into MAP2+mature neurons,and PF activated the WNT/β-catenin pathway to induce the differentiation of TH+dopaminergic neurons.XAV939 can block this effect.The TH+neurons in the PD model were less than those in the sham group(0.051±0.018 vs.1.000±0.350,t=4.705,P<0.010).After transplanting PF+NSCs into the striatum of PD rats,the number of rotations was less than that in the NSCs group(4.33±0.58 vs.6.33±0.58,t=4.240,P<0.05).Moreover,the number of TH+neurons increased(0.699±0.105 vs.0.151±0.101,t=6.532,P<0.05).Conclusion PF promotes the differentiation of NSCs into dopaminergic neurons and enhances their activity through the Wnt-β-catenin pathway,improves the motor function of PD rats,and provides a new strategy for cell replacement therapy.
作者
彭世君
王乐朋
欧阳佳
刘如恩
Peng Shijun;Wang Lepeng;Ouyang Jia;Liu Ru’en(Department of Neurosurgery,Peking University People’s Hospital,Beijing 100035,China)
出处
《中华实验外科杂志》
2025年第7期1310-1314,共5页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(82074174)
国家重点研发计划项目(2019YFE0117100)。
