脑络欣通方预处理的骨髓间充质干细胞外泌体促进氧糖剥夺再灌注神经干细胞增殖  

Bone Marrow Mesenchymal Stem Cell-derived Exosomes Pretreated with Naoluo Xintong Decoction Promote the Proliferation of Neural Stem Cells Under the Condition of Oxygen-Glucose Deprivation/Reoxygenation Injury

作　　者：刘明明 张涵智 江慧慧 赵雨彤 洪璐 陈卫东 何玲[3,4] 王妍妍[1] LIU Mingming;ZHANG Hanzhi;JIANG Huihui;ZHAO Yutong;HONG Lu;CHEN Weidong;HE Ling;WANG Yanyan(School of Pharmacy,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Key Laboratory of Chinese Medicinal Formula of Anhui Province,Anhui Hefei 230012,China;College of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Key Laboratory of Xin’an Medicine,Ministry of Education,Anhui Hefei 230038,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]中药复方安徽省重点实验室,安徽合肥230012 [3]安徽中医药大学中医学院,安徽合肥230012 [4]新安医学教育部重点实验室,安徽合肥230038

出　　处：《安徽中医药大学学报》2025年第1期102-108,共7页Journal of Anhui University of Chinese Medicine

基　　金：国家自然科学基金项目(82204671)。

摘　　要：目的观察脑络欣通方(Naoluo Xintong Decoction,NLXTD)预处理后的骨髓间充质干细胞外泌体(bone marrow mesenchymal stem cells derived exosomes,BMSCs-Exos)对氧糖剥夺再灌注(oxygen-glucose deprivation/reperfusion,OGD/R)损伤条件下C17.2小鼠神经干细胞(neural stem cells,NSCs)增殖的影响。方法利用全骨髓法提取原代小鼠BMSCs,采用流式细胞术对BMSCs的表面标志物进行鉴定;收集BMSCs细胞上清并用差速离心法分离Exos,之后运用透射电子显微镜、纳米颗粒跟踪分析和Western blot法进行鉴定;免疫荧光法鉴定C17.2小鼠NSCs,建立OGD/R损伤的NSCs模型,通过观察细胞状态以及利用EdU法检测NLXTD-Exos对NSCs的促增殖作用。结果提取的Exos呈典型茶托状结构。与对照组比较,模型组NSCs脱落较多,EdU阳性细胞率显著降低(P<0.05);与模型组比较,Exos组和NLXTD-Exos组细胞状态改善,漂浮细胞减少,EdU阳性细胞率显著升高(P<0.05);且NLXTD-Exos组的EdU阳性细胞率显著高于Exos组(P<0.05)。结论NLXTD预处理可提高BMSCs-Exos对OGD/R损伤的NSCs增殖的促进作用。Objective To investigate the effect of bone marrow mesenchymal stem cell-derived exosomes(BMSCs-Exos)pretreated with Naoluo Xintong Decoction(NLXTD)on the proliferation of mouse C17.2 neural stem cells(NSCs)under the condition of oxygen-glucose deprivation/reperfusion(OGD/R)injury.Methods The whole bone marrow method was used to extract primary mouse BMSCs,and flow cytometry was used to identify the surface markers of BMSCs.The supernatant of BMSCs was collected and exosomes(Exos)were isolated by differential centrifugation and were then identified by transmission electron microscopy,nanoparticle tracking analysis,and Western blot.Immunofluorescence assay was used to identify the mouse C17.2 NSCs,and a model of NSCs with OGD/R injury was established.Cellular state was observed and the EdU method was used to observe the effect of NLXTD-Exos in promoting the proliferation of NSCs.Results The Exos extracted showed a typical tea tray-like structure.Compared with the control group,the model group had an increase in the shedding of NSCs and a significant reduction in the rate of EdU-positive cells(P<0.05);compared with the model group,the Exos group and the NLXTD-Exos group showed a significant improvement in cellular state,a significant reduction in shed cells,and a significant increase in the rate of EdU-positive cells(P<0.05),and the NLXTD-Exos group had a significantly higher rate of EdU-positive cells than the Exos group(P<0.05).Conclusion NLXTD pretreatment can enhance the effect of BMSCs-Exos in promoting the proliferation of NSCs with OGD/R injury.

关 键 词：脑络欣通方 外泌体 氧糖剥夺再灌注 神经干细胞 

分 类 号：R285.5[医药卫生—中药学]