Involvement of miR-15b-5p in depression-like behavior in Parkinson's disease by down-regulating BDNF/TrkB/PSD95 expressions
10.3760/cma.j.cn115354-20240618-00359
- VernacularTitle:miR-15b-5p通过下调BDNF/TrkB/PSD95表达参与PD中抑郁样行为发生
- Author:
Hanjiang LUO
1
;
Chengli WU
;
Jinyong ZHU
;
Min CHEN
Author Information
1. 桂林医学院附属医院神经科学实验室，广西神经系统疾病临床医学研究中心，广西脑与认知神经科学重点实验室，桂林 541001
- Keywords:
Parkinson's disease;
Depression;
miR-15b-5p;
Brain-derived neurotrophic factor;
Synaptic plasticity
- From:
Chinese Journal of Neuromedicine
2024;23(7):649-658
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the mechanism of miR-15b-5p involving in depression-like behavior in Parkinson's disease (PD).Methods:(1) Eighteen C57BL/6N mice were randomly divided into PD group, intervention group and control group ( n=6). PD models in PD group were established by stereotaxically injecting 0.25 mg/kg rotenone into the right striatum; mice in the intervention group were injected with 0.25 mg/kg rotenone and miR-15b-5p inhibitor lentivirus, while mice in the control group were injected with equal volume of PBS. Four weeks after that, open field test and rotarod test were performed to evaluate the motor ability, and sucrose preference and forced swimming tests were performed to evaluate the depression-like behaviors. And then, proteins and miRNAs in the substantia nigra were extracted; real-time fluorescent quantitative reverse transcription PCR (qRT-PCR) was used to detect the miR-15b-5p expression,and Western blotting and immunofluorescent staining were used to detect the tyrosine hydroxylase (TH), brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), and postsynaptic density protein 95 (PSD95) protein expressions. (2) The 100 nmol/L miR-15b-5p mimic/inhibitor and their negative control sequences were transfected into SH-SY5Y cells on 6-well plates (named miR-15b-5p mimic group, miR-15b-5p mimic control group, miR-15b-5p inhibitor group and miR-15b-5p inhibitor control group, respectively); 48 h after that, BDNF, TrkB and PSD95 protein expressions were detected by Western blotting and miR-15b-5p expression by qRT-PCR. (3) The 100 ng BDNF 3'-UTR wild-type or mutant luciferase reporter vector plasmids and 100 nmol/L miR-15b-5p mimic/inhibitor or their negative control sequences were co-transfected into SH-SY5Y cells on 24-well plates, and luciferase reporter activity assay was performed 48 h after co-transfection to detect the luciferase activity. Results:(1) Compared with the control group, the PD group had significantly reduced movement speed, shortened rotarod drop latency, decreased percentage of sucrose preference, and prolonged immobility time ( P<0.05); compared with the PD group, the intervention group had significantly increased movement speed, prolonged rotarod drop latency, increased percentage of sucrose preference, and shortened immobility time ( P<0.05). (2) Compared with the miR-15b-5p mimic control group, the miR-15b-5p mimic group had significantly increased miR-15b-5p expression, and decreased BDNF, TrkB and PSD95 protein expressions (100.00±5.75 vs. 66.79±5.90; 100.00±5.95 vs. 84.46±5.77; 100.00±7.02 vs. 80.43±3.25, P<0.05). Compared with the miR-15b-5p inhibitor control group, the miR-15b-5p inhibitor group had significantly decreased miR-15b-5p expression, and increased BDNF, TrkB and PSD95 expressions (100.00±6.81 vs. 119.90±5.66; 100.00±2.88 vs. 110.10±4.15; 100.00±2.19 vs. 124.60±11.69, P<0.05). Compared with the control group, PD group had significantly increased miR-15b-5p expression, and significantly decreased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (100.00±9.20 vs. 63.60±12.80; 100.00±9.88 vs. 71.95±10.00; 100.00±5.16 vs. 70.37±8.43; 100.00±7.01 vs. 68.12±10.22; 100.00±12.99 vs. 48.23±12.58) in the substantia nigra ( P<0.05); compared with the PD group, the intervention group had significantly lower miR-15b-5p expression and increased TH, BDNF, TrkB and PSD95 expressions and BDNF fluorescent intensity (63.60±12.80 vs. 90.69±9.84; 71.95±10.00 vs. 93.31±4.50; 70.37±8.43 vs. 88.11±4.10; 68.12±10.22 vs. 89.59±5.93; 48.23±12.58 vs. 83.65±10.52) in the substantia nigra ( P<0.05). (3) Compared with the BDNF 3'-UTR wild-type+miR-15b-5p mimic control group, the BDNF 3'-UTR wild-type+miR-15b-5p mimic group had significantly decreased luciferase activity (100.00±5.07 vs. 90.59±1.75, P<0.05); compared with the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor control group, the BDNF 3'-UTR wild-type+miR-15b-5p inhibitor group had significantly increased luciferase activity (100.00±5.08 vs. 152.20±31.87, P<0.05). Conclusion:MiR-15b-5p is involved in depression-like behavior in PD by down-regulating the BDNF/TrkB/PSD95 expressions.
