The neuroprotective role of tissue inhibitor of metalloproteinase-2 in MPP+- or 6-OHDA-treated SK-N-BE(2)C and SH-SY5Y human neuroblastoma cells

doi:10.1016/j.neulet.2009.10.084

Neuroscience Letters

Volume 468, Issue 2, 4 January 2010, Pages 136-140

https://doi.org/10.1016/j.neulet.2009.10.084 Get rights and content

Abstract

Tissue inhibitors of metalloproteinases (TIMPs) are endogenous inhibitors of matrix metalloproteinases (MMPs), and the aberrant expressions of MMPs are strongly associated with neuroinflammation and neuronal cell death. In the present study, we found that two well-known dopaminergic neurotoxins, MPP⁺ and 6-OHDA, reduced TIMP-2 expression at the mRNA and protein levels in two human neuroblastoma cell lines (SK-N-BE(2)C and SH-SY5Y). To investigate the role of TIMP-2, these cells were transfected with TIMP-2 expression plasmid and viabilities were compared after treating cells with MPP⁺ or 6-OHDA. It was found that TIMP-2 overexpression attenuated the cell deaths induced by MPP⁺ or 6-OHDA, and that the degree of protection conferred was greater for MPP⁺-treated cells. Furthermore, the introduction of TIMP-2 siRNA into SK-N-BE(2)C cells aggravated the cell deaths induced by MPP⁺ or 6-OHDA. These findings collectively show that endogenously expressed TIMP-2 has a neuroprotective role, and they imply that the inhibition of TIMP-2 expression by MPP⁺ or 6-OHDA may contribute, in part, to neuronal cell death. These findings suggest that TIMP-2 expressional enhancement provides a potential therapeutic strategy for the treatment of neurodegenerative diseases such as Parkinson's disease.

Section snippets

Acknowledgement

This work was supported by a grant from Neurobiology Research Program (M10412000010-05N1200-01000) funded by the Ministry of Science and Technology, Republic of Korea.

References (22)

D. Blum et al.
Molecular pathways involved in the neurotoxicity of 6-OHDA, dopamine and MPTP: contributing to the apoptotic theory in Parkinson's disease
Prog. Neurobiol.
(2001)
K. Brew et al.
Tissue inhibitors of metalloproteinases: evolution, structure and function
Biochim. Biophys. Acta
(2000)
W.S. Choi et al.
Characterization of MPP(+)-induced cell death in a dopaminergic neuronal cell line: role of macromolecule synthesis, cytosolic calcium, caspase, and Bcl-2-related proteins
Exp. Neurol.
(1999)
M.L. Corcoran et al.
Tissue inhibitor of metalloproteinase-2 stimulates fibroblast proliferation via a cAMP-dependent mechanism
J. Biol. Chem.
(1995)
J. Dzwonek et al.
Matrix metalloproteinases and their endogenous inhibitors in neuronal physiology of the adult brain
FEBS Lett.
(2004)
N. Fager et al.
Differential spatial distribution and temporal regulation of tissue inhibitor of metalloproteinase mRNA expression during rat central nervous system development
Mech. Dev.
(2000)
S.Y. Kim et al.
Curcumin is a potent broad spectrum inhibitor of matrix metalloproteinase gene expression in human astroglioma cells
Biochem. Biophys. Res. Commun.
(2005)
E. Lambert et al.
TIMPs as multifacial proteins
Crit. Rev. Oncol./Hematol.
(2004)
K.J. Leco et al.
Murine tissue inhibitor of metalloproteinases-4 (Timp-4): cDNA isolation and expression in adult mouse tissues
FEBS Lett.
(1997)
D. Leppert et al.
Matrix metalloproteinases: multifunctional effectors of inflammation in multiple sclerosis and bacterial meningitis
Brain Res. Brain Res. Rev.
(2001)

S. Lorenzl et al.

Tissue inhibitors of matrix metalloproteinases are elevated in cerebrospinal fluid of neurodegenerative diseases

J. Neurol. Sci.

(2003)

Cited by (12)

Minocycline inhibits ICAD degradation and the NF-κB activation induced by 6-OHDA in PC12 cells
2014, Brain Research
Citation Excerpt :
The results of 6-OHDA studies suggest that oxidative stress-mediated apoptotic cell death is a major contributing factor to the selective degeneration of dopaminergic neurons in this PD model (Glinka and Youdim, 1995). It has been shown that 6-OHDA impairs mitochondrial dysfunction and caspase activation in a variety of cell lines and primary neuronal cultures (Kim et al., 2010; Kupershmidt et al., 2010; Liu et al., 2007; Puttonen et al., 2008; Yoon et al., 2011). Minocycline (7-dimethylamino-6-dimethyl-6-deoxytetracycline) is a second-generation, semi-synthetic tetracycline analogue that has been used for over 30 years (Yong et al., 2004) against both Gram-positive and Gram-negative bacteria.
6-Hydroxydopamine (6-OHDA) is a neurotoxin that is commonly employed to induce lesions of the dopaminergic pathways to generating experimental models of Parkinson’s disease (PD) in rodents. Antioxidant and anti-inflammatory therapy approaches have been the focus of attention in the treatment of neurodegenerative. PD and Alzheimer’s diseases, and oxidative stress have been implicated in these diseases. In this study, we investigated the neuroprotective effects of minocycline and the signalling pathway that is possibly involved in a PC12 cell model of PD. The results indicated that 6-OHDA cytotoxicity was accompanied by an increment in lactate dehydrogenase (LDH) release, an increase in caspase-3 protein activity, an increase in ROS generation, MDA content and decrease in the SOD, CAT activities and cell viability. Moreover, treatment with 6-OHDA alone for 24 h resulted in ICAD degradation, increased nuclear translocation of NF-κB, and increased p53 expression. However, pretreatment with minocycline (5, 10, 20 µM) for 24 h significantly reduced LDH release, reduced caspase-3 protein production, reduced ROS production, MDA content and attenuated the decrease in SOD, CAT activities and cell viability. Additionally, minocycline (20 µM) markedly decreased the levels of cleaved ICAD protein, down-regulated p53 activity and inhibited the nuclear translocation of NF-κB. The neuroprotective effects of minocycline were attributable to its potent antioxidant activities, which prevented the nuclear translocation of NF-κB and the subsequent promotion of cell death. Therefore, the present study supports the notion that minocycline may be a promising neuroprotective agent for the treatment of Parkinson’s disease.
Trasferrin receptor 2 gene regulation by microRNA 221 in SH-SY5Y cells treated with MPP<sup>+</sup> as Parkinson's disease cellular model
2013, Neuroscience Research
Citation Excerpt :
All the experiments were performed in triplicate. Evaluation of cell viability under MPP+ treatment Although several authors report different MPP+ concentrations to treat SH-SY5Y to create an in vitro model of Parkinson's disease, the most commonly used concentration is of 0.5 mM (Kim et al., 2010). In order to evaluate the toxicity due to the use of this drug, we performed a viability (MTS) assay, testing two different concentrations of MPP+ after 24, 48 and 72 h.
Parkinson's disease (PD) is one of the most frequent human neurodegenerations. The neurodegeneration in PD is related to cellular iron increase but the mechanisms involved in iron accumulation remain unclear. Transferrin receptor type 2 (TFR2) is a protein expressed on cell membrane and involved in the cellular iron uptake. We hypothesized that microRNA 221 could regulate the expression of TfR2 in an in vitro model of Parkinson's disease, SH-SY5Y cells treated with MPP⁺.
The miRNA 221 was selected by in silico analysis of several miRNAs predicted to target the TFR2 gene in SHSY5Y cells treated with MPP⁺. Taqman miRNA assay was used to evaluate the expression of the selected miRNAs. Using a luciferase assay we demonstrated the inhibition of TFR2 by miRNA 221.
We show that in PD cellular model, TFR2 expression is regulated by miRNA 221. TFR2 and miR 221 are inversely correlated in SHSY5Y cells during the treatment with MPP⁺. Moreover, overexpression of miRNA 221 decreases the expression of TFR2, respectively, at the mRNA and protein levels. The inhibition of endogenous miRNA 221 also is able to regulate TFR2.
These data suggest that miRNA 221 regulate TFR2 in PD model.
Thiamine and Parkinson's disease
2012, Journal of the Neurological Sciences
Citation Excerpt :
MMP-9 is also elevated in MPTP-induced parkinsonism in mice [120]. The application of dopaminergic neurotoxins to two human neuroblastoma cell lines downregulates the transcription and translation of tissue inhibitor of metalloproteinase (TIMP)-2, effectively enhancing MMP activity [121]. Exendin-4, which is an analog of glucagon-like peptide 1 (GLP-1), significantly attenuates the loss of SN neurons and the striatal dopaminergic fibers in the MPTP-induced PD model, and inhibits the expression of MMP-3 [122].
Parkinson's disease (PD) is the second most common form of neurodegeneration in the elderly population. PD is clinically characterized by tremors, rigidity, slowness of movement and postural imbalance. A significant association has been demonstrated between PD and low levels of thiamine in the serum, which suggests that elevated thiamine levels might provide protection against PD. Genetic studies have helped identify a number of factors that link thiamine to PD pathology, including the DJ-1 gene, excitatory amino acid transporters (EAATs), the α-ketoglutarate dehydrogenase complex (KGDHC), coenzyme Q10 (CoQ10 or ubiquinone), lipoamide dehydrogenase (LAD), chromosome 7, transcription factor p53, the renin–angiotensin system (RAS), heme oxygenase-1 (HO-1), and poly(ADP-ribose) polymerase-1gene (PARP-1). Thiamine has also been implicated in PD through its effects on L-type voltage-sensitive calcium channels (L-VSCC), matrix metalloproteinases (MMPs), prostaglandins (PGs), cyclooxygenase-2 (COX-2), reactive oxygen species (ROS), and nitric oxide synthase (NOS).
Recent studies highlight a possible relationship between thiamine and PD. Genetic studies provide opportunities to determine which proteins may link thiamine to PD pathology. Thiamine can also act through a number of non-genomic mechanisms that include protein expression, oxidative stress, inflammation, and cellular metabolism. Further studies are needed to determine the benefits of using thiamine as a treatment for PD.
Dopaminergic neurotoxicant 6-OHDA induces oxidative damage through proteolytic activation of PKCδ in cell culture and animal models of Parkinson's disease
2011, Toxicology and Applied Pharmacology
Citation Excerpt :
Results from MPTP and 6-OHDA studies suggest that oxidative stress-mediated apoptotic cell death is a major contributing factor to the selective degeneration of dopaminergic neurons in the PD model. 6-OHDA has been shown to impair mitochondrial dysfunction and caspase activation in a variety of cell lines and primary neuronal cultures (Choi et al., 1999; Dodel et al., 1999; Elkon et al., 2004; Jordan et al., 2004; Kim et al., 2010; Kupershmidt et al., 2010; Liu et al., 2007; Puttonen et al., 2008; Yoon et al., 2010). However, the role of cell signaling molecules downstream of caspases in the degeneration of dopaminergic neurons in PD is yet to be identified.
The neurotoxicant 6-hydroxydopamine (6-OHDA) is used to investigate the cellular and molecular mechanisms underlying selective degeneration of dopaminergic neurons in Parkinson's disease (PD). Oxidative stress and caspase activation contribute to the 6-OHDA-induced apoptotic cell death of dopaminergic neurons. In the present study, we sought to systematically characterize the key downstream signaling molecule involved in 6-OHDA-induced dopaminergic degeneration in cell culture and animal models of PD. Treatment of mesencephalic dopaminergic neuronal N27 cells with 6-OHDA (100 μM) for 24 h significantly reduced mitochondrial activity and increased cytosolic cytochrome c, followed by sequential activation of caspase-9 and caspase-3. Co-treatment with the free radical scavenger MnTBAP (10 μM) significantly attenuated 6-OHDA-induced caspase activities. Interestingly, 6-OHDA induced proteolytic cleavage and activation of protein kinase C delta (PKCδ) was completely suppressed by treatment with a caspase-3-specific inhibitor, Z-DEVD-FMK (50 μM). Furthermore, expression of caspase-3 cleavage site-resistant mutant PKCδ^D327A and kinase dead PKCδ^K376R or siRNA-mediated knockdown of PKCδ protected against 6-OHDA-induced neuronal cell death, suggesting that caspase-3-dependent PKCδ promotes oxidative stress-induced dopaminergic degeneration. Suppression of PKCδ expression by siRNA also effectively protected N27 cells from 6-OHDA-induced apoptotic cell death. PKCδ cleavage was also observed in the substantia nigra of 6-OHDA-injected C57 black mice but not in control animals. Viral-mediated delivery of PKCδ^D327A protein protected against 6-OHDA-induced PKCδ activation in mouse substantia nigra. Collectively, these results strongly suggest that proteolytic activation of PKCδ is a key downstream event in dopaminergic degeneration, and these results may have important translational value for development of novel treatment strategies for PD.
TIMP2 ameliorates blood-brain barrier disruption in traumatic brain injury by inhibiting Src-dependent VE-cadherin internalization
2024, Journal of Clinical Investigation
Metalloproteinases and their tissue inhibitors in Alzheimer’s disease and other neurodegenerative disorders
2019, Cellular and Molecular Life Sciences

View all citing articles on Scopus

View full text

The neuroprotective role of tissue inhibitor of metalloproteinase-2 in MPP+- or 6-OHDA-treated SK-N-BE(2)C and SH-SY5Y human neuroblastoma cells

Abstract

Section snippets

Acknowledgement

Prog. Neurobiol.

Biochim. Biophys. Acta

Exp. Neurol.

J. Biol. Chem.

FEBS Lett.

Mech. Dev.

Biochem. Biophys. Res. Commun.

Crit. Rev. Oncol./Hematol.

FEBS Lett.

Brain Res. Brain Res. Rev.

J. Neurol. Sci.

The neuroprotective role of tissue inhibitor of metalloproteinase-2 in MPP⁺- or 6-OHDA-treated SK-N-BE(2)C and SH-SY5Y human neuroblastoma cells