摘要
目的探讨姜黄素改善帕金森病(PD)小鼠行为缺陷的作用机制。方法建立1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的PD小鼠模型,采用转棒、爬杆和旷场实验评估小鼠的运动能力,通过免疫荧光染色观察脑黑质部位酪氨酸羟化酶(TH)阳性神经元数量,采用实时荧光定量逆转录聚合酶链反应法检测中脑部位肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β和IL-6等炎症因子mRNA表达水平,采用蛋白质印迹法检测核因子-κB(NF-κB)和核苷酸结合寡聚结构域样受体蛋白3(NLRP3)通路相关基因蛋白表达水平。结果成功建立小鼠PD模型,伴有明显运动障碍,表现转棒维持时间缩短、爬杆时间延长趋势、旷场内总运动距离和中心区运动时间明显缩短;模型小鼠黑质部位TH阳性神经元数量减少(P<0.05);中脑部位相比对照出现神经炎症,表现为促炎因子TNF-α、IL-1β、IL-6、NF-κB的mRNA表达水平升高(均P<0.05),NF-κB、磷酸化NF-κB、NLRP3、半胱氨酸蛋白酶-1(Caspase-1)蛋白表达水平升高(均P<0.05)。80 mg/kg姜黄素治疗模型小鼠的行为缺陷相比未干预的模型小鼠得到明显改善,表现为转棒维持时间明显升高(P<0.05),爬杆时间明显减少,旷场内总运动距离和中心区域活动时间明显升高(P<0.05);免疫荧光显示黑质部位TH阳性DA能神经元数量显著增加(P<0.05);中脑部位TNF-α、IL-1β和NF-κB的mRNA的表达水平明显降低(均P<0.05);NF-κB、NLRP3和Caspase-1蛋白表达水平下降(均P<0.05)。40 mg/kg姜黄素治疗模型小鼠与未干预的模型小鼠各项指标比较差异均无统计学意义(均P>0.05)。结论姜黄素可以改善PD小鼠模型的行为缺陷,逆转黑质部位TH阳性神经元损伤,80 mg/kg姜黄素治疗效果优于40 mg/kg,可能通过降低脑内神经炎症因子表达来实现。
Objective To explore the mechanism by which curcumin improves behavioral deficits in a mouse model of Parkinson's disease(PD).Methods A PD mouse model was established using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).The motor abilities of the mice were assessed using the rotarod,pole,and open-field tests.The number of tyrosine hydroxylase(TH)-positive neurons in the substantia nigra was observed using immunofluorescence staining.The mRNA expression levels of inflammatory factors such as tumor necrosis factorα(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)in the midbrain were detected using quantitative real-time reverse transcription polymerase chain reaction.The protein expression levels of genes related to the nuclear factor-κB(NF-κB)and nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)pathways were measured using Western blotting.Results The PD mouse model was successfully established,with significant motor impairments,including shortened rotarod retention time,prolonged pole test time,and decreased total distance traveled and time spent in the center of the open field.The number of TH-positive neurons in the substantia nigra of the model mice was reduced(P<0.05).Compared with the control group,the midbrain of the model mice exhibited neuroinflammation,characterized by increased mRNA expression levels of pro-inflammatory factors TNF-α,IL-1β,IL-6,and NF-κB(all P<0.05),as well as elevated protein expression levels of NF-κB,p-NF-κB,NLRP3,and Caspase-1(all P<0.05).Compared with non-treated model mice,curcumin treatment at 80 mg/kg significantly improved behavioral deficits in the model mice,with increased rotarod retention time(P<0.05),decreased pole test time,and increased total distance traveled and time spent in the center of the open field(P<0.05).Immunofluorescence revealed a significant increase in the number of TH-positive dopaminergic neurons in the substantia nigra(P<0.05).The mRNA expression levels of TNF-α,IL-1β,and NF-κB in the midbrain were significantly reduced(all P<0.05),and the protein expression levels of NF-κB,NLRP3 and Caspase-1 were decreased(all P<0.05).In contrast,curcumin treatment at 40 mg/kg did not show significant differences in any of the measured parameters compared with the untreated model mice(all P>0.05).Conclusion Curcumin can improve behavioral deficits in the PD mouse model and reverse the damage to TH-positive neurons in the substantia nigra.The therapeutic effect of 80 mg/kg curcumin is superior to that of 40 mg/kg,and this effect may be achieved by reducing the expression of neuroinflammatory factors in the brain.
作者
李文惠
赵志弘
王莉娟
何金晶
刘玉婷
韩秋琴
LI Wenhui;ZHAO Zhihong;WANG Lijuan;HE Jinjing;LIU Yuting;HAN Qiuqin(Collaborative Innovation Center,Shanghai University of Medicine&Health Sciences,Shanghai 201318,China)
出处
《浙江医学》
2025年第12期1233-1239,I0003,共8页
Zhejiang Medical Journal
基金
上海市“科技创新行动计划”启明星培育(扬帆专项)(23YF1418200)
上海市卫生健康委员会卫生行业临床研究专项(青年项目)(20234Y0294)。
关键词
帕金森病小鼠
姜黄素
运动缺陷
神经炎症
Mice with Parkinson's disease
Curcumin
Motor deficits
Neuroinflammation
作者简介
通信作者:韩秋琴,E-mail:hqq@sumhs.edu.cn。
