摘要
目的:探讨虎杖苷(PD)定向诱导骨髓间充质干细胞(BMSCs)向神经分化的潜能及其联合干细胞移植治疗脊髓损伤(SCI)的机制。方法:提取、培养并鉴定小鼠BMSCs细胞,CCK-8检测细胞活性,分组诱导后通过免疫荧光染色,观察PD对BMSCs向神经分化的影响。建立SCI动物模型,随机分组后干预,检测脊髓伤区神经元标志物蛋白及mRNA表达水平,组织形态学变化、行为学评分等,评估PD灌胃联合细胞移植治疗SCI小鼠的疗效。结果:与标准诱导组比较,PD联合诱导能提高BMSCs向神经元样细胞分化的效率,PD联合诱导组细胞中神经元标志物MAP-2、NeuN表达水平显著高于标准诱导组(P<0.05),其中30μmol/L浓度时效果最明显(P<0.05)。动物实验中,细胞移植组、细胞移植+PD灌胃组均能有效缓解脊髓伤区病理改变,上调神经元标志物蛋白、mRNA的表达,促进神经丝跨越胶质瘢痕区生长,并改善动物后肢活动功能,该趋势在细胞移植+PD灌胃组中更显著(P<0.05)。结论:PD通过促进移植BMSCs在体内向神经元样细胞分化,促进轴突修复,改善SCI小鼠后肢运动功能从而发挥治疗作用。
Objective:To explore the potential of polydatin(PD)in inducing the neural differentiation of bone marrow mesenchymal stem cells(BMSCs),and to investigate the mechanism of combined stem cell transplantation therapy in treating spinal cord injury(SCI).Methods:Mouse BMSCs were isolated,cultured and identified.Cell viability was detected by the CCK-8 assay.The PD-promoted neural differentiation of BMSCs was observed by immunofluorescent staining.The moderate SCI model was established and treated after randomly grouping.Protein and mRNA levels of neuronal markers at lesion site were evaluated.The therapeutic effect of PD gavage combined with BMSCs transplantation on SCI mice was assessed by observing the histomorphological changes and behavioral outcomes.Results:Compared with the standard induction group,PD improved the differentiation efficiency of BMSCs into neuron-like cells.The expression levels of neuronal markers MAP-2,NeuN in the combined induction group was significantly higher than those in the standard induction group(P<0.05),with the most significant effect at the concentration of 30μmol/L(P<0.05).In animal experiment,both interventions could effectively alleviate the pathological changes in spinal cord tissues,up-regulate the protein and mRNA expressions of neuronal markers,promote the growth of neurofilament across the glial scar,and improve hindlimb motor function,which were more significant in the PD gavage combined with BMSCs transplantation group(P<0.05).Conclusion:PD exerts therapeutic effect by enhancing the differentiation of transplanted BMSCs into neuron-like cells in vivo,promoting axonal repair,and improving locomotor function of SCI mice.
作者
詹吉恒
栾继耀
罗丹
李兴
侯永辉
侯宇
陈树东
肖志锋
林定坤
ZHAN Ji-heng;LUAN Ji-yao;LUO Dan;LI Xing;HOU Yong-hui;HOU Yu;CHEN Shu-dong;XIAO Zhi-feng;LIN Ding-kun(The Second Clinical Medical College,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405,China;department of Spinal Surgery,Guangdong Provincial Hospital of Chinese Medicine,Guangzhou 510120,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2020年第5期2628-2633,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81673992,No.81704095)
广东省科技计划项目(No.2016 A020226008)
广东省自然科学基金项目(No.2019A1515010323)。
关键词
虎杖苷
骨髓间充质干细胞
神经分化
脊髓损伤
轴突修复
中医经络再生
神经元样细胞
机制
Polydatin
Bone marrow mesenchymal stem cells
Neural differentiation
Spinal cord injury
Axon repair
Neural regeneration in traditional Chinese medicine
Neuron-like cells
Mechanism
