Nuclear orphan receptor Nur77 promotes α-synuclein degradation in Parkinson's disease by autophagy
10.13699/j.cnki.1001-6821.2018.10.018
- VernacularTitle:核孤儿受体Nur77促进α-突触核蛋白自噬性降解在帕金森病中的作用
- Author:
Rui SAI
1
;
Jun-Qiang YAN
;
Jian-Nan WU
;
Meng-Li GUO
;
Shuo WU
;
Liang QIAO
;
Li-Na HUANG
Author Information
1. 河南科技大学临床医学院
- Keywords:
nuclear orphan receptor Nur77;
α-synuclein;
1-methyl-4-pehny1-pyridine
- From:
The Chinese Journal of Clinical Pharmacology
2018;34(10):1199-1202
- CountryChina
- Language:Chinese
-
Abstract:
Objective To demonstrate the nuclear orphan receptor Nur77 agonist fungal polyketone (Cytosporone B,Csn-B) protects the neurons in Parkinson's disease cell model.Methods The SH-SY5Y cells were divided into blank group,control group (Csn-B),model group (MPP +) and experimental group (MPP + + Csn-B).The relative viability of human neuroblastoma cells SH-SY5Y induced by 1-methyl-4-pehny1-pyridine (MPP+) was measured with MTT assay.The expression of microtubule associated protein 1 light chain3 (LC3)-Ⅱ / Ⅰ and α-synuclein protein was detected by Western blotting.The expression of LC3 mRNA and α-synuclein mRNA were detected by qPCR.Immunofluorescence was used to observe the expression of LC3 and α-synuclein in each group.Results The MTT results showed that MPP + significantly inhibited proliferation of SH-SY5Y cells in a dose dependent manner.Western blotting showed that,LC3-Ⅱ and LC3-Ⅰ gray scale in blank group,control group,model group and experimental group were 0.21 ±0.02,0.23 ±0.03,0.32 ±0.03,0.69 ±0.05,LC3-Ⅱ/Ⅰ in model group and experimental group was significantly higher than that in blank group(P ＜ 0.01),and LC3-Ⅱ/Ⅰ in experimental group was significantly higher than that in model group (P ＜ 0.01).The α-synuclein in blank group,control group,model group and experimental group were 0.30 ± 0.05,0.51 ± 0.06,2.18 ± 0.12,0.73 ± 0.08,the expression of α-synuclein in model group and experimental group were significantly higher than that in blank group (P ＜0.01).The expression of α-synuclein protein in experimental group was significantly lower than that in model group (P ＜ 0.01).The relative expression of LC3 mRNA in blank group,control group,model group and experimental group were 3.94 ±0.18,4.04 ±0.13,6.01 ±0.21,7.43 ±0.35,the LC3 mRNA in model group and experimental group were significantly higher than that in blank group (P ＜ 0.01),LC3 mRNA in experimental group was significantly higher than that in model group (P ＜0.01).The α-synuclein mRNA in blank group,control group,model group and experimental group were 3.09 ±0.19,3.47 ±0.24,7.18 ±0.30,4.60 ±0.27,the expression of α-synuclein mRNA in model group and experimental group were significantly higher than that in blank group (P ＜0.01).The expression of α-synuclein mRNA in experimental group was significantly lower than that in model group (P ＜0.01).Immunofluorescence showed that LC3 protein was mainly located in cytoplasm.The expression of LC3 in model group and experimental group were significantly higher than that in blank group,and the expression level of LC3 in experimental group was higher than that in model group.oα-synuclein protein was distributed evenly in the cytoplasm and nucleus.The expression of α-synuclein in model group and experimental group was higher than that in blank group,experimental group was lower than that in model group.Conclusion In Parkinson' s disease cell model,Nur77 agonist can promote the degradation of α-synuclein,the mechanism may be related to increase autophagy level.
