Mechanism Study on LPIN1/PPARA Alleviating the Progression of Parkinson's Disease in Rats by Inhibiting SLC47A1-Mediated Ferroptosis of Neurons
10.3969/j.issn.1671-7414.2024.04.012
- VernacularTitle:LPIN1/PPARA通过抑制SLC47A1介导的神经元铁死亡缓解帕金森病模型大鼠病情进展的机制研究
- Author:
Mingyue CAO
1
;
Wei WANG
;
Meining ZHOU
Author Information
1. 大庆龙南医院/齐齐哈尔医学院第五附属医院检验科,黑龙江大庆 163453
- Keywords:
Parkinson's disease;
LPIN1;
PPARA;
SLC47A1;
ferroptosis
- From:
Journal of Modern Laboratory Medicine
2024;39(4):63-71
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of lipin1(LPIN1)on the progression of Parkinson's disease(PD)in rats and the possible molecular mechanism of its regulation.Methods The PD rat model was established by injection of 6-Hydroxydopamine hydrobromide(6-OHDA)into the medial forebrain tract of rats,and the LPIN1-overexpressing adenovirus was stably transfected to evaluate the behavioral changes of rats.The content of Fe2+and Glutathione(GSH)and the protein level of tyrosine hydroxylase(TH)in rat brain were detected,and HE staining was used to observe the pathological changes in rat brains.The PD cell model was constructed in vitro,and TH,α-synuclein(α-syn),LPIN1 protein levels and cell viability were detected.LPIN1 small interfering siRNA sequence and overexpression vector and peroxisome proliferator-activated receptor(PPARA)small interfering(siRNA)and overexpression vector were transfected,or ferroptosis inducer erastin was used to treat cell for 24 h,then cells were treated with 6-OHDA for 48h.The levels of Fe2+,reactive oxygen species(ROS),malondialdehyde(MDA),GSH and inflammatory factors in cells were detected to evaluate ferroptosis.Cell viability was detected with CCK-8,and the expressions of ferroptosis related proteins were detected with Western blot.The interacting protein PPARA of LPIN1 was predicted by STRING database and verified by Co-IP analysis.The binding site of PPARA to the promoter of solute carrier family 47 member 1(SLC47A1)was predicted by JASPAR bioinformatics and verified by Ch-IP analysis.Results The fur of the rats in the model group was frightened,and PD symptoms such as continuous tremor,slow movement and weakened activity were shown.The motor behavior and PD symptoms of LPIN1 group were improved/alleviated compared with the model group.Compared with the sham operation group,the total distance of the model group was shortened,the average speed was reduced,and the step length was reduced,while the total resting time was prolonged,the step width was widened,and the gait variation rate was increased,and the differences were significant(t=4.470～26.556,all P＜0.05).Compared with sham operation group,Fe2+content in brain tissue of model group was increased,while GSH content and TH protein expression were decreased,with significance differences(t=8.305,13.305,7.709,all P＜0.05).Compared with the model group,the behavioral evaluation,the level of indexes and the pathological changes of brain tissue in LPIN1 group were improved/alleviated.In addition,6-OHDA decreased PC-12 cell viability,reduced the levels of TH and LPIN1 protein,and increased the level of α-syn protein in a dose-dependent manner,and the differences were significant(F=31.023,7.350,9.124,15.841,all P＜0.05).Silencing LPIN1 intensified the inhibitory effect of 6-OHDA on the viability of PC-12 cells(t=2.209,P＜0.05),and overexpression of LPIN1 could counteract the effect of 6-OHDA.Overexpression of Lpin1 decreased the secretion of IL-1β and IL-6,increased the protein levels of SLC47A1 and GPX4,decreased the levels of Fe2+,MDA and ROS,and increased GSH content(t=3.013～11.639,all P＜0.05).Erastin reversed the inhibitory effect of Lpin1 overexpression on ferroptosis,and reduced the viability of PC-12 cells(t=3.087～7.581,all P＜0.05).LPIN1 interacted with PPARA protein and promoted PPARA expression,while PPARA bound to SLC47A1 promoter and promoted SLC47A1 transcriptional activation.Overexpression of PPARA counteracted the effect of Lpin1 silencing on PC-12 cells.Conclusion Overexpression of LPIN1 may reduce neuronal cell apoptosis by inhibiting ferroptosis mediated by PPARA/SLC47A1 axis,thus alleviating the progression of PD model rats.
