CAJ | 학술논문

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／mL）处理24 h，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。
목적：탐토작약감대α－돌촉핵단백적강해도경－자서화범소－단백매체통로적영향。방법：PC12세포용혹불용 MPP＋（0．5 mmol／L）처리24 h，연후용작약감（50μmol／L）혹뢰파매소（0．2μg／mL）처리24 h，MTT법검측세포증식활성，Western인적검측α－돌촉핵단백、미관상관단백경련3（LC3-Ⅱ）화E1적단백질표체수평，공취초현미경관찰α－돌촉핵단백화LC3-Ⅱ재세포내적형광강도급공정위정황。결과：（1）여MPP＋조상비，뢰파매소화작약감처리후세포과양화경매화초양화물기화매활력균현저하강（P ＜0．001），세포형태회복정상；（2）MPP＋처리도치 LC3－Ⅱ화E1표체수평승고，뢰파매소처리후LC3－Ⅱ증고이E1수평하강，작약감처리기현저상조LC3－Ⅱ（자서）야상조료 E1적표체（범소－단백매체도경）（P ＜0．001），촉진료α－돌촉핵단백적강해；（3）MPP＋증강세포내α－돌촉핵단백화LC3-Ⅱ적형광강도，작약감처리후α－돌촉핵단백적형광강도하강。결론：본연구결과표명작약감동시현저상조자서화범소단백매체도경，촉진료α－돌촉핵단백적강해，감소세포손상。
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 cells 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-Ⅱand E1. MPP+ promoted the increase ofLC3-Ⅱ but inhibited E1. 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 immunofluorescence signal of intracellular α-synuclein and LC3. Fluorescence intensity of α-synuclein decreasedafter PF treatment. Conclusion PF may significantly upregulate both autophagy and ubiquitin proteasome pathways, promote the degradation of α-synuclein and reduce cell damage. These findings suggest Paeoniflorin may be a potential therapy for neurodegenerative diseases.