Mir-141-3p Regulates Apoptosis and Mitochondrial Membrane Potential via Targeting Sirtuin1 in a 1-Methyl-4-Phenylpyridinium in vitro Model of Parkinson’s Disease

Abstract

Objectives. Parkinson’s disease (PD) is a common neurodegenerative disease characterized by the loss of midbrain dopaminergic neurons in the substantia nigra. The present study investigated miR-141-3p/sirtuin1 (SIRT1) activity in a 1-methyl-4-phenylpyridinium- (MPP+-) induced PC12-cell model of PD. Methods. PC12 cells were exposed to MMP+ following induction of differentiation by nerve growth factor (NGF). miR-141-3p and SIRT1 expressions were examined using RT-qPCR and western blot. Cell viability was evaluated using the MTT assay. Apoptosis percentage, reactive oxygen species (ROS) production, and mitochondrial membrane potential (Δψm) were evaluated using flow cytometry. Expression of Nuclear factor-kappa B- (NF-κB-) related proteins was determined by western blot. Bioinformatic analysis, RT-qPCR, and luciferase reporter assay were used to confirm the interaction between miR-141-3p and SIRT1. Results. miR-141-3p was upregulated, and SIRT1 was downregulated in MPP+-treated PC12 cells. MPP+ treatment also upregulated nitric oxide synthase 1 (Nos1) and α-synuclein. miR-141-3p induced apoptosis, oxidative stress, mitochondrial dysfunction, and downregulated the SIRT1 mRNA expression. The luciferase reporter assay showed that SIRT1 was the target of miR-141-3p. SIRT1 transfection attenuated apoptosis, ROS production and maintained Δψm. SIRT1 also downregulated Nos1, tumor necrosis factor-α (TNF-α), interleukin 1 beta (IL-1β), interleukin 6(IL-6) and upregulated B cell lymphoma 2 (Bcl-2) protein. In addition, SIRT1 activator resveratrol blocked the effects of miR-141-3p mimic on Nos1, α-synuclein, and mitochondrial membrane potential. SIRT1 inhibitor sirtinol reversed the biological effects of miR-141-3p. Conclusion. Increased miR-141-3p induced apoptosis, oxidative stress, and mitochondrial dysfunction in MPP+-treated PC12 cells by directly targeting the SIRT1 expression. Our study provided a potential therapeutic strategy for PD.

Astrocytic MicroRNA in Ageing, Inflammation, and Neurodegenerative Disease

“…Further investigation confirmed the IL-1β-induced upregulation of miR-30d-5p and miR-141-3p in both ADEVs and astrocytes ( Gayen et al, 2020 ; Table 1 and Figures 1A,B ). Augmented miR-30d-5p has been shown to upregulate apoptosis and downregulate autophagy by respectively targeting SMAD2 ( Yu and Liu, 2020 ) and the autophagosome-associated protein Beclin 1 ( Zhao et al, 2017 ), while increased miR-141-3p, which downregulates the neuroprotective protein Sirtuin 1, was associated with increased apoptosis and mitochondrial dysfunction ( Zheng et al, 2020 ). Thus, similar effects could occur in both astrocytes and ADEV-ingesting cells ( Figures 1A,B ).…”

“…The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and metabolite MPP+ are taken up into neurons via dopamine transporters, resulting in selective degeneration of dopaminergic neurons accompanied by parkinsonian symptoms ( Kitayama et al, 1998 ) and increased expression of α-syn protein in differentiated PC12 cells ( Zheng et al, 2020 ). Interestingly, exposure of astrocytes to MPP+ induces astrogliosis and the production of pro-inflammatory cytokines ( Yu et al, 2016 ) and alters the miRNA cargo of ADEVs ( Shakespear et al, 2020 ; Table 1 ).…”

Crosstalk between regulatory non-coding RNAs and oxidative stress in Parkinson’s disease

“…Both microRNA-141-3p (miR-141-3p) and microRNA-9-5p (miR-9-5p) were found to target the 3’ UTR of SIRT1 mRNA. Since SIRT1 inhibited the formation of α-syn aggregates, knockdown of miR-141-3p and miR-9-5p may alleviate oxidative stress and boost the viability of Das (Wang Z. et al, 2019 ; Zheng et al, 2020 ). In addition, microglia are thought to have a role in the pathophysiology of PD, since cytotoxic substances released from activated microglia can exacerbate oxidative stress.…”

Revisiting the miR-200 Family: A Clan of Five Siblings with Essential Roles in Development and Disease

“…Interestingly, Delavar et al [ 75 ] showed that miR-141 also targets SIRT1 expression and correlates with PD-related pathogenic processes. Mechanistically, in a 1-methyl-4-phenylpyridinium- (MPP+-) induced in vitro PD model, the upregulation of miR-141-3p induced increased apoptosis, oxidative stress, and mitochondrial membrane potential through the direct targeting of SIRT1 expression [ 69 ]. The same study also shows that resveratrol (a SIRT1 activator) blocked and sirtinol (a SIRT1 inhibitor) reversed the abovementioned biological effects of miR-141-3p, respectively.…”