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

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.

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Changes in mitochondrial membrane potential during oxidative stress-induced apoptosis in PC12 cells

Journal of Neuroscience Research ◽

10.1002/(sici)1097-4547(19971101)50:3<413::aid-jnr7>3.0.co;2-l ◽

1997 ◽

Vol 50 (3) ◽

pp. 413-420 ◽

Cited By ~ 142

Author(s):

Takumi Satoh ◽

Yasushi Enokido ◽

Hitoshi Aoshima ◽

Yasuo Uchiyama ◽

Hiroshi Hatanaka

Keyword(s):

Oxidative Stress ◽

Membrane Potential ◽

Pc12 Cells ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Induced Apoptosis

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Thrombopoietin Protects Cardiomyocytes from Iron-Overload Induced Oxidative Stress and Mitochondrial Injury

Cellular Physiology and Biochemistry ◽

10.1159/000430173 ◽

2015 ◽

Vol 36 (5) ◽

pp. 2063-2071 ◽

Cited By ~ 11

Author(s):

Shing Chan ◽

Godfrey Chifung Chan ◽

Jieyu Ye ◽

Qizhou Lian ◽

Jianliang Chen ◽

...

Keyword(s):

Oxidative Stress ◽

Membrane Potential ◽

Iron Overload ◽

Protective Effect ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Lipid Hydroperoxides ◽

Ros Production ◽

H9c2 Cells ◽

Induced Apoptosis

Background/Aims: Thalassaemia accompanied with iron-overload is common in Hong Kong. Iron-overload induced cardiomyopathy is the commonest cause of morbidity and mortality in patients with β-thalassaemia. Chronic iron-overload due to blood transfusion can cause cardiac failure. Decreased antioxidant defence and increased ROS production may lead to oxidative stress and cell injury. Iron-overload may lead to heart tissue damage through lipid peroxidation in response to oxidative stress, and a great diversity of toxic aldehydes are formed when lipid hydroperoxides break down in heart and plasma. Methods: Iron entry into embryonic heart H9C2 cells was determined by calcein assay using a fluorometer. Reactive oxygen species (ROS) production in cells treated with FeCl3 or thrombopoietin (TPO) was monitored by using the fluorescent probe H2DCFDA. Changes in mitochondrial membrane potential of H9C2 cells were quantified by using flow cytometry. Results: We demonstrated that iron induced oxidative stress and apoptosis in cardiomyocytes, and that iron increased ROS production and reduced cell viability in a dose-dependent manner. Iron treatment increased the proportion of cells with JC-1 monomers, indicating a trend of drop in the mitochondrial membrane potential. TPO exerted a cardio-protective effect on iron-induced apoptosis. Conclusions: These findings suggest that iron-overload leads to the generation of ROS and further induces apoptosis in cardiomyocytes via mitochondrial pathways. TPO might exert a protective effect on iron-overload induced apoptosis via inhibiting oxidative stress and suppressing the mitochondrial pathways in cardiomyocytes.

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Bcl-2 protects against FCCP-induced apoptosis and mitochondrial membrane potential depolarization in PC12 cells

Biochimica et Biophysica Acta (BBA) - General Subjects ◽

10.1016/s0304-4165(99)00073-2 ◽

1999 ◽

Vol 1428 (2-3) ◽

pp. 357-371 ◽

Cited By ~ 72

Author(s):

Gwenda Dispersyn ◽

Rony Nuydens ◽

Rich Connors ◽

Marcel Borgers ◽

Hugo Geerts

Keyword(s):

Membrane Potential ◽

Pc12 Cells ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Membrane ◽

Induced Apoptosis

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Encircling granulosa cells protects against di-(2-ethylhexyl)phthalate-induced apoptosis in rat oocytes cultured in vitro

Zygote ◽

10.1017/s0967199419000121 ◽

2019 ◽

Vol 27 (4) ◽

pp. 203-213 ◽

Cited By ~ 1

Author(s):

Anima Tripathi ◽

Vivek Pandey ◽

A.N. Sahu ◽

Alok K. Singh ◽

Pawan K. Dubey

Keyword(s):

Oxidative Stress ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Granulosa Cells ◽

Mitochondrial Membrane ◽

Dependent Manner ◽

Apoptotic Markers ◽

Induced Apoptosis ◽

Ethylhexyl Phthalate

SummaryThe present study investigated if the presence of encircling granulosa cells protected against di(2-ethylhexyl)phthalate (DEHP)-induced oxidative stress in rat oocytes cultured in vitro. Denuded oocytes and cumulus–oocyte complexes (COCs) were treated with or without various doses of DEHP (0.0, 25.0, 50.0, 100, 200, 400 and 800 μM) in vitro. Morphological apoptotic changes, levels of oxidative stress and reactive oxygen species (ROS), mitochondrial membrane potential, and expression levels of apoptotic markers (Bcl2, Bax, cytochrome c) were analyzed. Our results showed that DEHP induced morphological apoptotic changes in a dose-dependent manner in denuded oocytes cultured in vitro. The effective dose of DEHP (400 µg) significantly (P>0.05) increased oxidative stress by elevating ROS levels and the mitochondrial membrane potential with higher mRNA expression and protein levels of apoptotic markers (Bax, cytochrome c). Encircling granulosa cells protected oocytes from DEHP-induced morphological changes, increased oxidative stress and ROS levels, as well as increased expression of apoptotic markers. Taken together our data suggested that encircling granulosa cells protected oocytes against DEHP-induced apoptosis and that the presence of granulosa cells could act positively towards the survival of oocytes under in vitro culture conditions and may be helpful during assisted reproductive technique programmes.

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Olfactomedin 4 Is Essential for Superoxide Production and Sensitizes Oxidative Stress-Induced Apoptosis in Neutrophils.

Blood ◽

10.1182/blood.v114.22.1356.1356 ◽

2009 ◽

Vol 114 (22) ◽

pp. 1356-1356

Author(s):

Wenli Liu ◽

Yueqin Liu ◽

Ruihong Wang ◽

Cuiling Li ◽

Chuxia Deng ◽

...

Keyword(s):

Oxidative Stress ◽

Membrane Potential ◽

Mitochondrial Membrane Potential ◽

Respiratory Chain ◽

Complex I ◽

Mitochondrial Membrane ◽

Mitochondrial Respiratory Chain ◽

H2o2 Production ◽

Induced Apoptosis ◽

Mitochondrial Respiratory

Abstract Abstract 1356 Poster Board I-378 Introduction Olfactomedin 4 (OLFM4), also called hGC-1, GW112 and pDP4, was first identified and specifically expressed in hematopoietic myeloid cells. OLFM4 expression in myeloid cells is regulated by transcription factors, PU1 and NF-κB. It has significant homology in its C-terminal domain with other olfactomedin-related proteins. OLFM4 encodes a 510 amino acid N-linked glycoprotein. The exact biological function of OLFM4, especially in neutrophils, is currently undefined. To characterize the in vivo function of OLFM4, we generated OLFM4 deficient mice (OLFM4-/-) and investigated its potential role in neutrophil functioins. Results 1) In this study, we showed that OLFM4 is a secreted glycoprotein and is also localized in the mitochondria, cytoplasm and cell membrane fractions of neutrophils. We demonstrated that OLFM4 interacts with GRIM-19 (Genes associated with Retinoid-IFN-induced Mortality-19), an apoptosis related protein, in the neutrophil mitochondria using co-immuoprecipitation assay. GRIM-19 is a subunit of complex I of mitochondrial respiratory chain and is essential for maintenance of mitochondrial membrane potential. Our result suggests that OLFM4 appears to be a novel component of complex I of mitochondrial respiratory chain and may be involved in regulation of mitochondrial membrane potential. 2) Mice heterozygous (OLFM4+/-) and homozygous (OLFM4-/-) for the null mutation in OLFM4 appeared to have normal development, fertility, and viability relative to wild-type (WT) mice. Whole blood analysis, differential leukocyte counts, blood chemistry and bone marrow smears were normal in OLFM4-/- mice, suggesting that OLFM4 is not essential for normal development and hematopoiesis in mice. 3) In response to LPS, fMLP and E.coli bacteria challenge, neutrophils from OLFM4-/- mice showed significantly reduced superoxide (O2−) and hydrogen peroxide (H2O2) production compared with WT mice. These results suggest that OLFM4 is an essential component to mediate O2− and H2O2 production in the neutrophil mitochondria under inflammation stimuli. 4) Exogenous H2O2 induced neutrophil apoptosis in a time and dose dependent manner in WT mice, but this induction of apoptosis was significantly reduced in OLFM4-/- mice. This result suggests that OLFM4 sensitizes and mediates H2O2-induced apoptosis in neutrophils. 5) Furthermore, we demonstrated that H2O2-stimulated mitochondrial membrane permeability reduction and caspase-3 and caspase-9 activation were inhibited in the neutrophils of OLFM4-/- mice. This result confirmed our hypothesis that OLFM4 may be involved in maintenance of mitochondrial membrane potential and suggests that OLFM4 may have opposite role as GRIM-19. 6) Moreover, Bax association with mitochondria and the cytoplasmic translocation of Omi/HtrA2 and Smac/DIABLO in response to H2O2 were inhibited in the neutrophils of OLFM4-/- mice. Conclusion Our results suggest: 1) OLFM4 has multiple subcellular localizations including mitochondria, cytoplasm, and cell membrane in neutrophils. The interaction of OLFM4 with GRIM-19 in the mitochondria suggests that OLFM4 is novel component of complex I of mitochondrial respiratory chain in the mitochondria of neutrophils, 2) OLFM4 is a novel mitochondrial molecule that is essential for O2− and H2O2 production in the neutrophils in the presence of inflammation stimuli, 3) Loss of OLFM4 in neutrophils does not trigger spontaneous apoptosis. However, OLFM4 sensitizes oxidative stress-induced apoptosis in mouse neutrophils. OLFM4 is involved in the regulation of mitochondria membrane potential and sensitizes cytoplasmic translocation of Omi/HtrA2 and Smac/DIABLO and caspases-3 and caspase-9 mediated apoptosis in the presence of oxidative stress. Disclosures No relevant conflicts of interest to declare.

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Effect of thyroid hormone on mitochondrial properties and oxidative stress in cells from patients with mtDNA defects

AJP Cell Physiology ◽

10.1152/ajpcell.00415.2007 ◽

2009 ◽

Vol 296 (2) ◽

pp. C355-C362 ◽

Cited By ~ 32

Author(s):

Keir J. Menzies ◽

Brian H. Robinson ◽

David A. Hood

Keyword(s):

Oxidative Stress ◽

Membrane Potential ◽

Thyroid Hormone ◽

Protein Expression ◽

Mitochondrial Membrane Potential ◽

Mitochondrial Biogenesis ◽

Mitochondrial Membrane ◽

Ros Production ◽

Mitochondrial Mass ◽

Mtdna Mutations

Mitochondrial (mt)DNA mutations contribute to various disease states characterized by low ATP production. In contrast, thyroid hormone [3,3′,5-triiodothyronine (T3)] induces mitochondrial biogenesis and enhances ATP generation within cells. To evaluate the role of T3-mediated mitochondrial biogenesis in patients with mtDNA mutations, three fibroblast cell lines with mtDNA mutations were evaluated, including two patients with Leigh's syndrome and one with hypertrophic cardiomyopathy. Compared with control cells, patient fibroblasts displayed similar levels of mitochondrial mass, peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), mitochondrial transcription factor A (Tfam), and uncoupling protein 2 (UCP2) protein expression. However, patient cells exhibited a 1.6-fold elevation in ROS production, a 1.7-fold elevation in cytoplasmic Ca2+ levels, a 1.2-fold elevation in mitochondrial membrane potential, and 30% less complex V activity compared with control cells. Patient cells also displayed 20–25% reductions in both cytochrome c oxidase (COX) activity and MnSOD protein levels compared with control cells. After T3 treatment of patient cells, ROS production was decreased by 40%, cytoplasmic Ca2+ was reduced by 20%, COX activity was increased by 1.3-fold, and ATP levels were elevated by 1.6-fold, despite the absence of a change in mitochondrial mass. There were no significant alterations in the protein expression of PGC-1α, Tfam, or UCP2 in either T3-treated patient or control cells. However, T3 restored the mitochondrial membrane potential, complex V activity, and levels of MnSOD to normal values in patient cells and elevated MnSOD levels by 21% in control cells. These results suggest that T3 acts to reduce cellular oxidative stress, which may help attenuate ROS-mediated damage, along with improving mitochondrial function and energy status in cells with mtDNA defects.

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Protective Effect of SCUBE1 on Oxidative Stress-induced Apoptosis in Human Ovarian Granulosa Cells

10.21203/rs.3.rs-285634/v1 ◽

2021 ◽

Author(s):

Huijiao Fu ◽

Xuzi Cai ◽

Qiwen Liu ◽

Wei Yang ◽

xuefeng wang

Keyword(s):

Membrane Potential ◽

Western Blot ◽

Cell Viability ◽

Mitochondrial Membrane Potential ◽

Granulosa Cells ◽

Mitochondrial Membrane ◽

Caspase 3 ◽

Cell Counting ◽

Ovarian Granulosa Cells ◽

Induced Apoptosis

Abstract Background: Apoptosis of ovarian granulosa cells (GCs) is a sign of follicular atresia. This study aimed to explore the role and mechanism of signal peptide, CUB domain, epidermal growth factor-like protein1 (SCUBE1) in protecting GCs from apoptosis induced by hydrogen peroxide (H2O2). Methods: Firstly, the expression of SCUBE1 on the ovaries of humans and mice was analyzed by qRT-PCR, western blot and immunohistochemistry. Subsequently, the H2O2 treated GCs were pretreated with SCUBE1 recombinant protein, and their cell viability and proliferation were detected by Cell Counting Kit-8 (CCK-8) assay. The levels of reactive oxygen species (ROS) and mitochondrial membrane potential (ΔΨm) in the cells were determined by DCFH-DA and rhodamine 123, respectively. The percentage of apoptotic cells was analyzed by flow cytometry after staining with Annexin V/PI. The expression levels of pathway related proteins, such as Bcl-2, Bax, p53, caspase-3, were determined by western blot analysis. Finally, the pathogenicity of SCUBE1 (c.1169C>G, p.P390R) were analyzed based on the software.Results: SCUBE1 was expressed in women of all ages and had the highest expression level in the ovaries in multiple organs and tissues of KM mouse. In vitro cell experiments show that SCUBE1 pretreatment reduced H2O2-induced apoptosis and improved cell viability. SCUBE1 also blocked the production of ROS in cells and improved mitochondrial membrane potential. After SCUBE1 pretreatment, anti-apoptotic protein Bcl-2 expression was upregulated, whereas the expression of the pro-apoptotic proteins Bax, Bax/Bcl-2, Caspase-3, and p53 were downregulated. Analysis of the impact of SCUBE1 (c.1169C >G, p.P390R) mutation from the aspect of mutation pathogenicity; protein stability; and gene haplotype insufficiency, indicated that the p.P390R mutation is significantly pathogenic.Conclusions: This is the first time that the potential role of SCUBE1 in protecting GCs from H2O2-induced damage through the mitochondrial pathways, attributing to POI, is studied. SCUBE1 (c.1169C >G, p.P390R) mutation has significant pathogenicity but the specific harm needs to be confirmed by further studies. Trial registration: Not applicable.

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Oxidative stress and mitochondrial membrane potential are involved in the cytotoxicity of perfluorododecanoic acid to neurons

Toxicology and Industrial Health ◽

10.1177/0748233720957534 ◽

2020 ◽

Vol 36 (11) ◽

pp. 892-897

Author(s):

Xuemei Fang ◽

Xingtao Zhang ◽

Hongxia Li

Keyword(s):

Oxidative Stress ◽

Membrane Potential ◽

Pc12 Cells ◽

Mitochondrial Membrane Potential ◽

Cognitive Deficits ◽

Mitochondrial Membrane ◽

Caspase 3 ◽

Oxygen Species ◽

Total Antioxidant ◽

The Brain

Perfluorododecanoic acid (PFDoA), used in numerous commercial products, was recently demonstrated to accumulate in the brain more easily than other perfluorinated compounds and to cause cognitive deficits. In this study, pheochromocytoma 12 (PC12) cells were exposed to doses of PFDoA to explore the cytotoxicity of this compound to neurons. The results showed that treatment with PFDoA decreased PC12 cell viability dose-dependently. Treatment with 50 and 100 µM PFDoA significantly increased reactive oxygen species ( p < 0.01) and malondialdehyde ( p < 0.01) and decreased total antioxidant capacity ( p < 0.05 and p < 0.01, respectively) in PC12 cells. The administration of 50 and 100 µM PFDoA led to a loss of mitochondrial membrane potential (MMP) ( p < 0.05 and p < 0.01, respectively) in PC12 cells. The activity of caspase 3 was obviously increased ( p < 0.05) in 100 µM PFDoA-treated PC12 cells. In general, the results demonstrated that PFDoA exposure could result in the disruption of MMP, which may contribute to the increase of oxidative stress and activation of the apoptotic signaling cascade in PC12 cells.

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