芍药苷对MPP+/MPTP诱导的帕金森病模型小鼠的保护作用
Protective effect of paeoniflorin on MPP+/MPTP-induced Parkinsonian mice
陆瑶 1鲁婷婷 1郑克迪 1秦鹏 1郑梅竹2
作者信息
- 1. 长春师范大学生命科学学院,吉林长春 130032
- 2. 长春师范大学中心实验室,吉林长春 130032
- 折叠
摘要
帕金森病(Parkinson's disease,PD)是常见的神经退行性疾病,已严重影响老年人的身体健康.为探究芍药苷(peoniflorin)抗帕金森病作用的活性及作用机制,本试验首先建立了 1-甲基-4-苯基-吡啶离子(1-methyl-4-phenylpyr-idinium,MPP+)诱导PC12帕金森细胞模型.通过细胞形态学观察、检测细胞活性、乳酸脱氢酶(LDH)的释放量、细胞内Ca2+浓度、活性氧(ROS)水平以及细胞凋亡检测,对芍药苷的神经保护作用进行初步的活性评价.再利用1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)致 PD 小鼠模型为研究对象,通过检测自主活动和滚轴能力表现,对芍药苷抗PD的作用进一步验证.在此基础上,通过Western blot检测相关凋亡蛋白Bcl-2、Bax、Caspase-3的表达水平研究其作用机制.结果显示,与正常对照组比较,模型组细胞数量减少,细胞体积明显缩小;细胞活性显著降低(P＜0.01);MPP+显著增加PC12细胞LDH释放率(P＜0.01)、ROS水平(P＜0.01)和Ca2+浓度(P＜0.01),同时细胞凋亡率显著增加(P＜0.01).与模型组比较,药物保护组细胞损伤减弱,活细胞数量增多,细胞活性增强;25、50 μmol/L剂量芍药苷保护组显著降低MPP+诱导的PC12细胞LDH释放率(P＜0.01)、ROS水平(P＜0.01)和Ca2+浓度(P＜0.01),显著缓解了 MPP+诱导的细胞凋亡(P＜0.01);MPTP诱导可显著减少小鼠的自主活动次数(P＜0.01)和滚轴时间(P＜0.01),经芍药苷预处理后显著增加小鼠自发站立次数(P＜0.05)和滚轴时间(P＜0.01);25、50 μmol/L芍药苷保护组显著增加了细胞内Bcl-2/Bax表达(P＜0.01),同时降低了Caspase-3的表达(P＜0.01).结果表明,芍药苷对MPP+和MPTP诱导帕金森病体内、外模型均具有神经保护作用,其作用机制是通过增加细胞内Bcl-2/Bax表达,降低了 Caspase-3的表达实现的.
Abstract
Parkinson's disease(PD)is a common neurodegenerative disease,which has seriously af-fected the physical health of elderly people.To investigate the anti-PD activity and mechanism of peoniflorin,1-methyl-4-phenylpyridinium(MPP+)-induced PC12 Parkinson's cell model was es-tablished in this study.Cell activity,release of lactatedehydrogenase(LDH),intracellular Ca2+con-centration,levels of reactive oxygen species(ROS)and apoptosis were detected by morphology observation.To evaluate the neuroprotective effect of paeoniflorin,autonomic dysfunction and roll-er ability were detected,and anti-PD effect of paeoniflorin was verified using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)-induced PD mouse model.On this basis,the expression levels of the proteins related to apoptosis,Bcl-2,Bax and Caspase-3,were detected by Western blot.The results showed that compared with the normal control group,the model group showed a decrease in the number of cells and a significant reduction in the cell body;the cell activity was significantly reduced(P＜0.01);MPP+significantly increased the rate of LDH release(P＜0.01),the level of ROS(P＜0.01),and the concentration of Ca2+ of PC12 cells,while the rate of apoptosis was significantly increased(P＜0.01);compared with the model group,the drug-protected group showed attenuated cell damage,increased number of viable cells,and enhanced cell activity.The level of MPP+-induced LDH release from PC12 cells(P＜0.01),ROS level(P＜0.01),and Ca2+concentration(P＜0.01)significantly decreased in 25 μmol/L and 50 μmol/L paeoniflorin-protected group,and it significantly mitigated MPP+-induced apop-tosis(P＜0.01);MPTP induction could significantly reduce the number of voluntary activities(P＜0.01)and rolling time(P＜0.01),and significantly increase the number of spontaneous standing(P＜0.05)and rolling time(P＜0.01)in mice pretreated with paeoniflorin.Mice in 50 μmol/L paeoniflorin-protected group significantly increased the expression of intracellular Bcl-2/Bax(P＜0.01),and simultaneously decreased the Caspase-3 expression(P＜0.01).In conclu-sion,paeoniflorin exerted neuroprotective effects on both in vivo and ex vivo models of MPP+and MPTP-induced PD,and its mechanism of action was realized by increasing the intracellular Bcl-2/Bax expression and decreasing the expression of Caspase-3.
关键词
芍药苷/帕金森/MPP+/MPTP/凋亡/小鼠Key words
paeoniflorin/Parkinson's disease/MPP+/MPTP/apoptosis/mice引用本文复制引用
基金项目
吉林省自然科学基金资助项目(20210101031JC)
出版年
2024
NETL
NSTL
万方数据