C2神经酰胺对帕金森病模型鼠多巴胺能神经元的保护作用  

Protective effect of C2 ceramide on dopaminergic neurons in a mouse model of Parkinson’s disease

作　　者：李家慧 齐雪 朱远峰 禹璐 刘立峰[1] 王鹏[1,2] Li Jiahui;Qi Xue;Zhu Yuanfeng;Yu Lu;Liu Lifeng;Wang Peng(Department of Human Anatomy,Beihua University,Jilin 132013,Jilin Province,China;Laboratory of Neurodegenerative Diseases,School of Basic Medical Sciences,Beihua University,Jilin 132013,Jilin Province,China)

机构地区：[1]北华大学基础医学院人体解剖学教研室,吉林省吉林市132013 [2]北华大学基础医学院神经退行性疾病研究室,吉林省吉林市132013

出　　处：《中国组织工程研究》2024年第11期1653-1659,共7页Chinese Journal of Tissue Engineering Research

基　　金：吉林省自然科学基金(YDZJ202201ZYTS575),项目负责人:王鹏;吉林省卫生与健康技术创新项目(2018J083),项目负责人:王鹏;北华大学研究生创新项目(北华研创合字[2021]054),项目负责人:李家慧。

摘　　要：背景:C2神经酰胺可减少Alpha突触核蛋白(Alpha-Synuclein,α-Syn)寡聚体的形成,其作为蛋白磷酸酶2A激动剂在中枢神经系统对细胞老化具有重要调节作用。目的:探究C2神经酰胺对多巴胺能神经元的保护作用机制。方法:将25只C57BL/6系小鼠随机分为对照组、模型组及C2神经酰胺低、中、高剂量组,每组5只。除对照组之外,其他各组均通过左侧纹状体注射突变型A53Tα-Syn寡聚体建立帕金森病小鼠模型,在纹状体注射第30天后,3个C2神经酰胺各剂量组小鼠一次性经胃灌注溶于生理盐水中的各剂量C2神经酰胺(1,5,10μg/g),对照组及模型组同法灌注等量生理盐水,于造模后第30-90天,连续每天给药,共60 d,期间观察各组小鼠行为学变化。在纹状体注射第90天后,在适度麻醉条件下对各组小鼠灌注取脑,以免疫组织化学染色分析小鼠中脑黑质多巴胺能神经元的变化,以ELISA法检测小鼠中脑α-Syn寡聚化和磷酸化水平,并分析影响α-Syn磷酸化的相关酶活性变化。结果与结论:①C2神经酰胺对经纹状体注射的突变型A53Tα-Syn寡聚体所导致的小鼠帕金森病样运动障碍具有改善作用,且C2神经酰胺高剂量组对帕金森病模型鼠运动障碍的改善作用更为明显(P<0.01);②突变型A53Tα-Syn寡聚体致使小鼠中脑黑质多巴胺能神经元数目明显减少(P<0.01),而C2神经酰胺高剂量组黑质多巴胺能神经元数目则明显增多(P<0.01);③与模型组相比,C2神经酰胺高剂量组小鼠中脑内α-Syn寡聚体和磷酸化α-Syn水平明显降低(P<0.01),而神经酰胺的水平增高(P<0.05),蛋白磷酸酶2A活力明显上调(P<0.01);④上述数据说明,C2神经酰胺可通过改善小鼠中脑组织α-Syn的磷酸化修饰环境,缓解了由寡聚的α-Syn所诱导的神经毒性作用,保护了小鼠中脑黑质多巴胺能神经元数量的减少,从而减轻帕金森病的运动障碍程度。BACKGROUND:C2 ceramide reduces the formation of Alpha-Synuclein(α-Syn)oligomers as the protein phosphatase 2A agonist,which has an important regulatory effect on cell aging in the central nervous system.OBJECTIVE:To investigate the protective mechanism of C2 ceramide on dopaminergic neurons.METHODS:Twenty-five C57BL/6 mice were randomly divided into control group,model group,C2 ceramide low-,medium-and high-dose groups(n=5 per group).Except for the control group,a mouse model of Parkinson’s disease was established by injecting mutant A53Tα-Syn oligomers into the left striatum in the other groups.On the 30th day after the striatal injection,three C2 ceramide groups were intragastrically administered with C2 ceramide(1,5,10μg/g)dissolved in saline at one time,while the control and model groups were administered with the same amount of saline within 30-90 days after modeling,for a total of 60 days.Behavioral changes in each group of mice were observed during this period.On the 90th day after striatal injection,mouse brain tissue was extracted by perfusion under anesthesia,and the changes of dopaminergic neurons in the midbrain substantia nigra were analyzed by immunohistochemical staining.The levels ofα-Syn oligomerization and phosphorylation in the midbrain of mice were detected by ELISA,and the changes of enzyme activities related toα-Syn phosphorylation were analyzed.RESULTS AND CONCLUSION:C2 ceramide had an ameliorating effect on Parkinson’s disease-like dyskinesia in mice caused by the striatal injection of mutant A53Tα-Syn oligomers.High-dose C2 ceramide showed better effects on dyskinesia in mice with Parkinson’s disease(P<0.01).The mutant A53Tα-Syn oligomers significantly reduced the number of dopaminergic neurons in the substantia nigra of mice(P<0.01),while the number of dopaminergic neurons in the substantia nigra increased significantly in the C2 ceramide high-dose group(P<0.01).The levels ofα-Syn oligomers and phosphorylatedα-Syn in the brain were significantly reduced in the C2 ceramide

关 键 词：C2神经酰胺 多巴胺能神经元 帕金森病 Alpha突触核蛋白 蛋白磷酸酶2A 寡聚体 磷酸化 保护作用 

分 类 号：R459.9[医药卫生—治疗学] R363[医药卫生—临床医学] R74