Blood plasma from patients with Parkinson's disease and multiple system atrophy can enhance the damage effect of α-synuclein aggregates on mitochondrial and lysosome functions
10.3760/cma.j.cn115354-20240814-00484
- VernacularTitle:帕金森病及多系统萎缩患者血浆增强α-突触核蛋白聚集体对细胞线粒体及溶酶体功能的影响
- Author:
Yu LUO
1
;
Chengli WU
1
;
Yong HU
1
;
Min CHEN
1
;
Hanjiang LUO
1
Author Information
1. 广西神经系统疾病临床医学研究中心，桂林医学院附属医院神经科学实验室，广西脑与认知神经科学重点实验室，桂林 541001
- Publication Type:Journal Article
- Keywords:
Parkinson's disease;
Multiple system atrophy;
α-synuclein;
Aggregate;
Mitochondria;
Lysosome
- From:
Chinese Journal of Neuromedicine
2024;23(12):1195-1204
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the effect of α-synuclein (α-Syn) aggregates incubated with plasma from patients with Parkinson's disease (PD) and multiple system atrophy (MSA) on mitochondrial and lysosomal functions, and further clarify the role of α-Syn in pathological mechanism of PD and MSA.Methods:Human neuroblastoma cells (SH-SY5Y) were treated with 10 μg/mL α-Syn aggregates incubated with phosphate buffered solution (PBS), and plasma from healthy controls (HCs), PD patients, and MSA patients, respectively; and the untreated SH-SY5Y cells were used as control group. Mito-Tracker Red CMXRos fluorescent staining was used to detect the mitochondrial membrane potential, Western blotting was used to detect the cytochrome C (Cyt C) expression in the mitochondria and cytoplasm, and adenosine triphosphate (ATP) test kit was used to detect the intracellular ATP level. In addition, the lysosomal associated membrane protein-1 (LAMP1) expression was detected by Western blotting, and cathepsin B activity was detected by Magic Red fluorescent staining.Results:(1) Effects of different α-Syn aggregates on mitochondrial function damage: compared with the control group, the PBS, HC, PD and MSA groups had significantly decreased relative fluorescent intensity (Mito-Tracker Red CMXRos staining) in the SH-SY5Y intracellular mitochondria ( P<0.05); the PD group and MSA group had significantly decreased relative fluorescent intensity compared with PBS group and HC group ( P<0.05); the MSA group had significantly decreased relative fluorescent intensity compared with the PD group (41.27%±5.97% vs. 60.05%±4.24%, P<0.05). Compared with the control group, the HC, PD and MSA groups had significantly decreased mitochondrial Cyt C protein expression, while the PBS, HC, PD and MSA groups had significantly increased cytoplasmic Cyt C protein expression ( P<0.05); the PD group and MSA group had significantly decreased mitochondrial Cyt C protein expression compared with the PBS group and HC group ( P<0.05); the MSA group had significantly increased cytoplasmic Cyt C protein expression compared with the PBS group ( P<0.05); the MSA group had significantly decreased mitochondrial Cyt C protein expression (51.50%±4.22%) and increased cytoplasmic Cyt C protein expression (199.40%±6.73%) compared with the PD group (65.52%±2.18% and 174.00%±16.18%, P<0.05). Compared with the control group, the PBS group, HC group, PD group and MSA group had significantly decreased ATP level ( P<0.05); the PD group and MSA group had significantly decreased ATP level compared with the PBS group and HC group ( P<0.05); the MSA group had significantly decreased ATP level (1.81±0.13 nmol/mg) compared with PD group (2.96±0.29 nmol/mg, P<0.05). (2) Effects of different α-Syn aggregates on lysosomal functions: compared with the control group, the PD group and MSA group had significantly decreased LAMP1 protein expression ( P<0.05); the MSA group had significantly decreased LAMP1 protein expression (60.74%±7.94%) compared with the PBS group, HC group, and PD group (79.70%±4.34%, P<0.05). Compared with the control group, the PBS, HC, PD and MSA groups had significantly enhanced relative fluorescent intensity of lysosomal cathepsin B ( P<0.05); the PD group and MSA group had significantly enhanced relative fluorescent intensity of lysosomal cathepsin B compared with PBS group and HC group ( P<0.05); the MSA group had significantly increased relative fluorescent intensity of lysosomal cathepsin B compared with PD group (399.00%±35.54% vs. 305.90%±24.87%, P<0.05). Conclusion:The α-Syn aggregates incubated with plasma from PD and MSA patients have damage to mitochondrial and lysosome functions, especially those incubated with plasma from MSA patients.
