小鼠胰腺癌细胞系Pan02来源外泌体有效改善帕金森病模型小鼠运动功能障碍OA
Murine pancreatic cancer cell line Pan02-derived exosomes effectively improve motor dysfunction in mouse models of Parkinson's disease
目的 探究小鼠胰腺癌细胞系 Pan02 源性外泌体(Exos)对帕金森病(PD)模型小鼠的治疗效应.方法 试剂盒法提取 Pan02 细胞源性外泌体,通过透射电镜(TEM)、纳米颗粒跟踪分析及 Western blot 进行鉴定表征.体外通过CCK-8法检测外泌体对 1-甲基-4-苯基吡啶(MPP+)诱导的小鼠中脑多巴胺能神经元细胞(MN9D)损伤模型的保护作用.基于 C57BL/6J 小鼠构建 1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的急性 PD 模型,设置野生型(WT)对照组、PD 模型组及外泌体(Exos)低、中、高剂量(Exo-L、Exo-M、Exo-H)组,尾静脉多次注射外泌体.通过平衡木、爬杆、转棒、旷场及 Y 迷宫实验评估小鼠运动功能及精神行为变化;免疫荧光染色检测黑质及纹状体多巴胺能神经元损伤情况;并构建荷瘤小鼠模型进行初步安全性评价.结果 外泌体可显著逆转 MPP+诱导的 MN9D 细胞活力下降(P＜0.001);与模型组相比,Exo-L 组平衡木穿越时间显著减少(P＜0.001);爬杆时间显著缩短(P＜0.05);Exo-M 组旷场自发运动总距离显著减少(P＜0.05);各组焦虑及认知相关指标差异无统计学意义;外泌体显著减轻MPTP 诱导的黑质酪氨酸羟化酶(TH)阳性神经元丢失(Exo-L 组P＜0.05,Exo-H 组P＜0.01)及纹状体TH 阳性纤维密度降低(Exo-L 组、Exo-M 组 P＜0.05).安全性评价显示,外泌体未促进荷瘤小鼠肿瘤生长.结论 Pan02 细胞源性外泌体可有效缓解 MPP+/MPTP 诱导的多巴胺能神经元损伤,改善 PD 模型小鼠运动功能障碍,且未表现出促瘤风险,具有良好的神经保护作用及安全性.
Objective To investigate the therapeutic effects of murine pancreatic cancer cell line Pan02l-derived exosomes(Exos)on Parkinson's disease(PD)in a mouse model.Methods Pan02 cell-derived exosomes were ex-tracted by commercially available kit and then characterized by transmission electron microscape(TEM),nanoparti-cle tracking analysis and Western blot.In vitro,the protective effect of exosomes on a mouse model of 1-methyl-4-phenylpyridinium(MPP+)-induced MN9D dopaminergic neuron injury was detected by CCK-8 assay.A C57BL/6J mice of acute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced PD was established.The mice were divided into a wild-type(WT)control group,a PD model group,and low,medium,and high-dose exosome groups(Exo-L,Exo-M,Exo-H).Exosomes were administered via multiple tail vein injections.Motor function and neurobehavioral changes were assessed using the balance beam test,pole test,rotarod test,open field test and Y-maze test.Damage to dopaminergic neurons in the substantia nigra and striatum was evaluated by immunofluores-cence microscopy.A tumor-bearing mouse model was also established for a preliminary safety evaluation.Results Exosomes markedly reversed the MPP+-induced decrease in MN9D cell viability(P＜0.001).Compared with the PD model group,the Exo-L group showed a substantial reduction in beam crossing time(P＜0.001)and pole de-scent time(P＜0.05).The Exo-M group exhibited a significant decrease in total distance moved in the open field test(P＜0.05).No notable differences were observed in anxiety and cognition-related behavioral parameters among all groups.Exosomes dramatically mitigated MPTP-induced loss of tyrosine hydroxylase(TH)-positive neurons in the substantia nigra(Exo-L group,P＜0.05;Exo-H group,P＜0.01)and reduction of TH-positive fiber density in the striatum(Exo-L and Exo-M groups,P＜0.05).Safety evaluation confirmed that exosomes did not promote tumor growth in tumor-bearing mice.Conclusions Pan02 cell-derived exosomes effectively alleviate MPP+/MPTP-induced dopaminergic neuron injury,ameliorate motor dysfunction in PD mouse models,and exhibit no tumor-pro-moting risk,demonstrating favorable neuro-protective effects and safety.
张可昕;肖芳艳;李文华;黄薇;刘雁勇;杨楠
中国医学科学院北京协和医学院 基础医学研究所 药理学系,北京 100005江西省药品不良反应监测中心,江西 南昌 330006中国医学科学院北京协和医学院 基础医学研究所 药理学系,北京 100005中国医学科学院北京协和医学院 基础医学研究所 药理学系,北京 100005中国医学科学院北京协和医学院 基础医学研究所 药理学系,北京 100005中国医学科学院北京协和医学院 基础医学研究所 药理学系,北京 100005
医药卫生
帕金森病1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)Pan02细胞外泌体
Parkinson's disease1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)Pan02 cellexosome
《基础医学与临床》 2026 (7)
957-964,8
中国医学科学院医学与健康科技创新工程(2021-I2M-1-020)
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