Abstract
Cypermethrin activates microglia, which is found to be decisive in neurodegeneration in the experimental rats. While the involvement of microglial activation in toxicant-induced neurodegeneration is reported, the effect of low concentration of cypermethrin on the expression of inflammatory proteins from the rat primary microglia is not yet properly understood. The study intended to delineate the effect of low concentration of cypermethrin on the expression and release of proteins from the microglia. Rat primary microglial cells were treated with cypermethrin to check the expression of inflammatory proteins. Cypermethrin-treated microglia conditioned media and cells were collected to measure the expression and release of inflammatory proteins. Cypermethrin augmented the protein kinase C-δ (PKC-δ), inducible nitric oxide synthase (iNOS), phosphorylated mitogen-activated protein kinase (MAPK) p38 and p42/44, matrix metalloproteinase (MMP)-3, and MMP-9 levels in the cell lysate and tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) levels in the microglia conditioned media. Pre-treatment with minocycline, a microglial activation inhibitor or rottlerin, a PKC-δ inhibitor, notably reduced the release of TNF-α in the conditioned media and expression of iNOS protein in the microglia. Minocycline reduced the expression of PKC-δ, phosphorylated p38 and p42/44 MAPKs, MMP-3, and MMP-9 proteins in the microglia. While cypermethrin-treated conditioned media induced the toxicity in the rat primary neurons, minocycline or rottlerin reduced the cypermethrin treated microglia conditioned media-induced toxicity. The outcomes of the present study suggest that cypermethrin activates microglia and releases TNF-α and IL-1β as well as up-regulates the expression of PKC-δ, iNOS, phosphorylated p38 and p42/44 MAPKs, MMP-3, and MMP-9 proteins, which could contribute to neurodegeneration.
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The data are available upon genuine request from the corresponding author.
References
Agrawal S, Singh A, Tripathi P et al (2015) Cypermethrin-induced nigrostriatal dopaminergic neurodegeneration alters the mitochondrial function: a proteomics study. Mol Neurobiol 51:448–465. https://doi.org/10.1007/s12035-014-8696-7
Block ML, Zecca L, Hong JS (2007) Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci 8:57–69. https://doi.org/10.1038/nrn2038
Burguillos MA, Deierborg T, Kavanagh E et al (2011) Caspase signalling controls microglia activation and neurotoxicity. Nature 472:319–324. https://doi.org/10.1038/nature09788
Choi DH, Kim EM, Son HJ et al (2008) A novel intracellular role of matrix metalloproteinase-3 during apoptosis of dopaminergic cells. J Neurochem 106:405–415. https://doi.org/10.1111/j.1471-4159.2008.05399.x
Garrido-Mesa N, Zarzuelo A, Gálvez J (2013) Minocycline: far beyond an antibiotic. Br J Pharmacol 169:337–352. https://doi.org/10.1111/bph.12139
Glass CK, Saijo K, Winner B et al (2010) Mechanisms underlying inflammation in neurodegeneration. Cell 140:918–934. https://doi.org/10.1016/j.cell.2010.02.016
Gottschall PE, Yu X (1995) Cytokines regulate gelatinase A and B (matrix metalloproteinase 2 and 9) activity in cultured rat astrocytes. J Neurochem 64:1513–1520. https://doi.org/10.1046/j.1471-4159.1995.64041513.x
Hirsch EC, Hunot S, Damier P et al (1998) Glial cells and inflammation in Parkinson’s disease: a role in neurodegeneration? Ann Neurol 44:S115–S120. https://doi.org/10.1002/ana.410440717
Kim YS, Kim SS, Cho JJ et al (2005) Matrix metalloproteinase-3: a novel signaling proteinase from apoptotic neuronal cells that activates microglia. J Neurosci 25(14):3701–3711. https://doi.org/10.1523/JNEUROSCI.4346-04.2005
Kim YS, Joh TH (2012) Matrix metalloproteinases new insights into the understanding of neurodegenerative disorders. Biomol Ther Seoul 20(133):143. https://doi.org/10.4062/biomolther.2012.20.2.133
Lawson LJ, Perry VH, Dri P, Gordon S (1990) Heterogeneity in the distribution and morphology of microglia in the normal adult mouse brain. Neuroscience 39:151–170. https://doi.org/10.1016/0306-4522(90)90229-w
Lee EJ, Ko HM, Jeong YH et al (2015) β-Lapachone suppresses neuroinflammation by modulating the expression of cytokines and matrix metalloproteinases in activated microglia. J Neuroinflammation 12:133. https://doi.org/10.1186/s12974-015-0355-z
Li Q, Barres BA (2018) Microglia and macrophages in brain homeostasis and disease. Nature Rev Immunol 18:225–242. https://doi.org/10.1038/nri.2017.125
Liberatore GT, Jackson-Lewis V, Vukosavic S et al (1999) Inducible nitric oxide synthase stimulates dopaminergic neurodegeneration in the MPTP model of Parkinson disease. Nat Med 5:1403–1409. https://doi.org/10.1038/70978
Lin L, Desai R, Wang X et al (2017) Characteristics of primary rat microglia isolated from mixed cultures using two different methods. J Neuroinflammation 14:101. https://doi.org/10.1186/s12974-017-0877-7
Lotharius J, Dugan LL, O’Malley KL (1999) Distinct mechanisms underlie neurotoxin-mediated cell death in cultured dopaminergic neurons. J Neurosci 19(4):1284–1293. https://doi.org/10.1523/JNEUROSCI.19-04-01284.1999
Lowry OH, Rosebrough NJ, Farr AL et al (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275 (PMID: 14907713)
Miller RL, Sun GY, Sun AY (2007) Cytotoxicity of paraquat in microglial cells: involvement of PKCδ-and ERK1/2-dependent NADPH oxidase. Brain Res 1167:129–139. https://doi.org/10.1016/j.brainres.2007.06.046
Mishra AK, Mishra S, Rajput C et al (2018) Cypermethrin activates autophagosome formation albeit inhibits autophagy owing to poor lysosome quality: relevance to Parkinson’s disease. Neurotox Res 33(2):377–387. https://doi.org/10.1007/s12640-017-9800-3
Mogi M, Harada M, Riederer P et al (1994) Tumor necrosis factor-α (TNF-α) increases both in the brain and in the cerebrospinal fluid from parkinsonian patients. Neurosci Lett 165:208–210. https://doi.org/10.1016/0304-3940(94)90746-3
Mogi M, Harada M, Narabayashi H et al (1996) Interleukin (IL)-1β, IL-2, IL-4, IL-6 and transforming growth factor-α levels are elevated in ventricular cerebrospinal fluid in juvenile Parkinsonism and Parkinson’s disease. Neurosci Lett 211:13–16. https://doi.org/10.1016/0304-3940(96)12706-3
Mun JY, Lee WY, Han SS (2005) Effects of cypermethrin on the dopaminergic neurons in the progressive hemiparkinsonian rats. Toxicol Mech Methods 15:399–404. https://doi.org/10.1080/15376520500194742
Nikodemova M, Duncan ID, Watters JJ (2006) Minocycline exerts inhibitory effects on multiple mitogen-activated protein kinases and IκBα degradation in a stimulus-specific manner in microglia. J Neurochem 96:314–323. https://doi.org/10.1111/j.1471-4159.2005.03520.x
Rosenberg GA (2009) Matrix metalloproteinases and their multiple roles in neurodegenerative diseases. Lancet Neurol 8:205–216. https://doi.org/10.1016/S1474-4422(09)70016-X
Rozovsky I, Finch CE, Morgan TE (1998) Age-related activation of microglia and astrocytes: in vitro studies show persistent phenotypes of aging, increased proliferation, and resistance to down-regulation. Neurobiol Aging 19:97–103. https://doi.org/10.1016/s0197-4580(97)00169-3
Saura J, Tusell JM, Serratossa J (2003) High-yield isolation of murine microglia by mild trypsinization. Glia 44(3):183–189. https://doi.org/10.1002/glia.10274
Saw S, Weiss A, Khokha R et al (2019) Metalloproteases: on the watch in the hematopoietic niche. Trends Immunol 40:1053–1070. https://doi.org/10.1016/j.it.2019.09.006
Schönwasser DC, Marais RM, Marshall CJ et al (1998) Activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase pathway by conventional, novel, and atypical protein kinase C isotypes. Mol Cell Biol 18:790–798. https://doi.org/10.1128/mcb.18.2.790
Scholz R, Sobotka M, Caramoy A et al (2015) Minocycline counter-regulates pro-inflammatory microglia responses in the retina and protects from degeneration. J Neuroinflammation 12:209. https://doi.org/10.1186/s12974-015-0431-4
Shen S, Yu S, Binek J et al (2005) Distinct signaling pathways for induction of type II NOS by IFNγ and LPS in BV 2 microglial cells. Neurochem Int 47:298–307. https://doi.org/10.1016/j.neuint.2005.03.007
Singh AK, Tiwari MN, Dixit A et al (2011) Nigrostriatal proteomics of cypermethrin-induced dopaminergic neurodegeneration: microglial activation-dependent and -independent regulations. Toxicol Sci 122:526–538. https://doi.org/10.1093/toxsci/kfr115
Singh AK, Tiwari MN, Prakash O et al (2012) A current review of cypermethrin-induced neurotoxicity and nigrostriatal dopaminergic neurodegeneration. Curr Neuropharmacol 10(1):64–71. https://doi.org/10.2174/157015912799362779
Singh AK, Tiwari MN, Upadhyay G et al (2012a) Long term exposure to cypermethrin induces nigrostriatal dopaminergic neurodegeneration in adult rats: postnatal exposure enhances the susceptibility during adulthood. Neurobiol Aging 33(2):404–415. https://doi.org/10.1016/j.neurobiolaging.2010.02.018
Tikka T, Fiebich BL, Goldsteins G et al (2001) Minocycline, a tetracycline derivative, is neuroprotective against excitotoxicity by inhibiting activation and proliferation of microglia. J Neurosci 21(8):2580–2588. https://doi.org/10.1523/JNEUROSCI.21-08-02580.2001
Toro R, Downward GS, Mark MVD et al (2019) Parkinson’s disease and long-term exposure to outdoor air pollution: a matched case-control study in the Netherlands. Environ Int 129:28–34. https://doi.org/10.1016/j.envint.2019.04.069
Weinert M, Selvakumar T, Tierney TS et al (2015) Isolation culture and long term maintenance of primary mesencephalic dopaminergic neurons from embryonic rodent brains. J Vis Exp 96:52475. https://doi.org/10.3791/52475
Wen J, Ribeiro R, Zhang Y (2011) Specific PKC isoforms regulate LPS-stimulated iNOS induction in murine microglial cells. J Neuroinflammation 8:38. https://doi.org/10.1186/1742-2094-8-38
Whitton PS (2007) Inflammation as a causative factor in the aetiology of Parkinson’s disease. Br J Pharmacol 150:963–976. https://doi.org/10.1038/sj.bjp.0707167
Woo MS, Park JS, Choi IY et al (2008) Inhibition of MMP-3 or -9 suppresses lipopolysaccharide-induced expression of proinflammatory cytokines and iNOS in microglia. J Neurochem 106(2):770–780. https://doi.org/10.1111/j.1471-4159.2008.05430.x
Zhang W, Liu HT (2002) MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res 12:9–18. https://doi.org/10.1038/sj.cr.7290105
Acknowledgements
We show our gratitude to the University Grants Commission, India, for endowing with the financial assistance to Saumya Mishra, the first author of the manuscript. The Council of Scientific and Industrial Research, India, granted the research fellowship to Charul Rajput. The CSIR-IITR communication number of this article is 3654.
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The first author, Saumya Mishra, designed, generated, acquired and analyzed the data and wrote the preliminary version of the manuscript. Charul Rajput extensively assisted Saumya Mishra in performing a few experiments as well as in data compilation and analysis. Mahendra Pratap Singh participated in the study design and exhaustively revised the lexis of the manuscript. All three authors have gone through the final version of the manuscript and agreed with its substance.
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Mishra, S., Rajput, C. & Singh, M.P. Cypermethrin Induces the Activation of Rat Primary Microglia and Expression of Inflammatory Proteins. J Mol Neurosci 71, 1275–1283 (2021). https://doi.org/10.1007/s12031-020-01753-y
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DOI: https://doi.org/10.1007/s12031-020-01753-y