芍药苷对α-突触核蛋白降解通路的影响  

The impact of Paeoniflorin on α-synuclein degradation pathway

作　　者：张永进[1] 王敏[1] 徐晶[1] 张謦芝 李秀明[1] 蔡增林[1] 

机构地区：[1]徐州医学院附属连云港医院神经内科,222002

出　　处：《实用医学杂志》2015年第19期3136-3139,共4页The Journal of Practical Medicine

基　　金：江苏省自然基金面上项目(编号:BK2011402)

摘　　要：目的:探讨芍药苷对α-突触核蛋白的降解途径-自噬和泛素-蛋白酶体通路的影响。方法:PC12细胞用或不用MPP+(0.5 mmol/L)处理24 h,然后用芍药苷(50μmol/L)或雷帕霉素(0.2μg/m L)处理24h,MTT法检测细胞增殖活性,Western印迹检测α-突触核蛋白、微管相关蛋白轻链3(LC3-Ⅱ)和E1的蛋白质表达水平,共聚焦显微镜观察α-突触核蛋白和LC3-Ⅱ在细胞内的荧光强度及共定位情况。结果:(1)与MPP+组相比,雷帕霉素和芍药苷处理后细胞过氧化氢酶和超氧化物歧化酶活力均显著下降(P<0.001),细胞形态恢复正常;(2)MPP+处理导致LC3-Ⅱ和E1表达水平升高,雷帕霉素处理后LC3-Ⅱ增高而E1水平下降,芍药苷处理既显著上调LC3-Ⅱ(自噬)也上调了E1的表达(泛素-蛋白酶体途径)(P<0.001),促进了α-突触核蛋白的降解;(3)MPP+增强细胞内α-突触核蛋白和LC3-Ⅱ的荧光强度,芍药苷处理后α-突触核蛋白的荧光强度下降。结论:本研究结果表明芍药苷同时显著上调自噬和泛素蛋白酶体途径,促进了α-突触核蛋白的降解,减少细胞损伤。Objective To study the impact of Paeoniflorin （PF） on α-synuclein degradation pathway. Methods PC12 cells were treated with or without MPP＋ （0.5mM） for 24 h, then treated with Paeoniflorin （50 uM） or Rapamycin （0.2 μg/ml） for 24 h. The proliferative activity of ceils was detected with the MTT method, and then the protein expression levels of α-synuclein, microtubule-associated protein light chain 3（LC3-Ⅱ） and E1 were detected by Western Blot. The expressions of α-synuclein and LC3 were detected by confocal microscopy. Results （1） CAT and SOD activity were significantly decreased after PF and RAPA treatment compared with MPP＋ （P 〈 0.001）. （2）MPP＋ activated both LC3-Ⅱ and El. MPP＋ promoted the increase of LC3-Ⅱ but inhibited El. PF significantly upregulated both LC3-Ⅱ （autophagy） and E1 expression （ubiquitin- proteasome pathway） （P 〈 0.001）, promoted degradation of α-synuclein, and reduced cell damage. （3） MPP＋ enhanced immunofluoreseenee signal of intraeellular α-synuelein and LC3. Fluorescence intensity of α-synuclein deereasedafter PF treatment. Conclusion PF may significantly upregulate both autophagy and ubiquitin proteasome pathways, promote the degradation of α-synuelein and reduce cell damage. These findings suggest Paeoniflorin may be a potential therapy for neurodegenerative diseases.

关 键 词：芍药苷 Α-突触核蛋白 泛素-蛋白酶体通路 自噬 

分 类 号：R741[医药卫生—神经病学与精神病学]