主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2020, Vol. 43 ›› Issue (12): 1027-1033.doi: 10.3969/j.issn.1006-2157.2020.12.009

• 中药药理 • 上一篇    下一篇

知母皂苷B-Ⅱ对MPP+,诱导的SH-SY5Y细胞帕金森病模型线粒体的保护作用*

张锦坤, 周梦琪, 马浩洁, 张宇昕, 程翠翠, 杨璐平, 盖聪, 高誉珊, 孙红梅#   

  1. 北京中医药大学中医学院 北京 100029
  • 收稿日期:2020-06-17 出版日期:2020-12-30 发布日期:2021-01-05
  • 通讯作者: #孙红梅,女,博士,教授,博士生导师,主要研究方向: 中医药防治脑病,E-mail: hm-sun@ 163.com
  • 作者简介:张锦坤,女,在读硕士生
  • 基金资助:
    *国家自然科学基金项目( No.81774110),北京中医药大学新教师启动基金项目(No.2020-JYB-XJSJJ-051)

Protective effects of timosaponin B-II on mitochondria of SH-SY5Y cell model of MPP+-induced Parkinson's disease*

Zhang Jinkun, Zhou Mengqi, Ma Haojie, Zhang Yuxin, Cheng Cuicui, Yang Luping, Gai Cong, Gao Yushan, Sun Hongmei#   

  1. School of Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China
  • Received:2020-06-17 Online:2020-12-30 Published:2021-01-05
  • Contact: Prof. Sun Hongmei, Ph. D.,Doctoral Supervisor. School of Chinese Medicine,Beijing University of Chinese Medicine. No.11 Beisanhuan Donglu Road, Chaoyang District, Beijing 100029. E-mail:hm-sun@163.com
  • Supported by:
    National Natural Science Foundation of China (No. 81774110)

摘要: 目的 探讨知母有效成分知母皂苷B-Ⅱ对1-甲基-4-苯基-吡啶离子(MPP+, ) 诱导的帕金森病细胞模型线粒体的保护作用。方法 选择人多巴胺能神经母细胞瘤(SH-SY5Y)细胞,用MPP +, 处理建立帕金森病细胞模型,分别采用终浓度为0.3、1.0、3.0 μmol/L 知母皂苷B-II干预。CCK-8法检测各组细胞存活率,流式细胞术检测细胞凋亡率、细胞活性氧(ROS)水平,激光扫描共聚焦显微镜拍照检测线粒体活性,Image J软件计算线粒体形态因子、网络分支平均长度及网络分支数,JC-1法检测线粒体膜电位,荧光素酶法检测细胞ATP水平。结果 与模型组比较,知母皂苷B-II低、中、高浓度组细胞存活率均上升(P<0.05或P<0.01)、凋亡率降低、线粒体活性提高,形态因子升高、网络分支平均长度及网络分支数增多,拮抗膜电位降低、相对ATP水平均上升、细胞ROS水平均升高(P<0.01),并且观察到知母皂苷B-II中浓度组在拮抗膜电位的降低、提高ATP水平作用更为明显。结论 知母皂苷B-II可改善MPP+, 对SH-SY5Y细胞线粒体功能的损伤,拮抗膜电位的降低、增加ATP生成、降低ROS产生,修复线粒体网络形态,从而减少细胞凋亡,达到治疗帕金森病的作用。

关键词: 知母皂苷B-Ⅱ, 神经母细胞瘤细胞, 帕金森病, 线粒体

Abstract: Objective To explore the protective effects of the effective component of timosaponin B-II (TB-II) on mitochondria in Parkinson’s disease (PD) model induced by 1-methyl-4-phenyl-pyridine (MPP+, ). Methods SH-SY5Y, a human dopaminergic neuroblastoma, was selected and treated with MPP+, to establish a PD cell model. TB-II at concentrations of 0.3, 1.0, and 3.0 μmol/L were used for intervention. CCK-8 was used to detect cell survival rate of different groups, flow cytometry cell apoptosis rate and reactive oxygen species (ROS) level, and laser scanning confocal microscope mitochondrial activity. Image J software was used to evaluate mitochondrial morphology factor, average length of network branch and number of network branches. JC-1 was employed to detect mitochondrial membrane potential, and luciferase cellular ATP level. Results Compared with the model group, the cell survival rate of the TB-II low, medium, and high concentration groups were significantly increased (P<0.05, P<0.01), the apoptosis rate was significantly reduced, the mitochondrial activity, the morphological factors, the average length and the number of network branchesantagonizing the decreace of membrane potential, the relative ATP level, the cellular ROS level were increased significantly (P<0.01). In addition, it was observed that the TB-II medium concentration group performed best in antagonizing the decrease of membrane potential and increasing ATP levels. Conclusion TB-II can limit MPP+, induced damage to SH-SY5Y cell mitochondrial function, antagonize the reduction of membrane potential, increase ATP production, reduce ROS production, repair mitochondrial network morphology, thereby suppressing apoptosis and relieving PD.

Key words: TB-II, SH-SY5Y cells, Parkinson's disease, mitochondria

中图分类号: 

  • R285.5