Fumaric Acids Do Not Directly Influence Gene Expression of Neuroprotective Factors in Highly Purified Rodent Astrocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Preparation and Culture of Highly Purified Astrocytes
2.2. RNA Isolation and Reverse Transcription Polymerase Chain Reaction (RT-PCR)
2.3. Statistical Analysis
3. Results
3.1. DMF is Biologically Active and DMF and MMF are not Toxic in Vitro
3.2. DMF and MMF have No Effect on Growth Factor Gene Expression in Highly Purified Activated Astrocytes
3.3. DMF and MMF have No Effect on Growth Factor and Cytokine Expression in Lipopolysaccharide Stimulated Astrocytes
4. Discussion
5. Limitations of the Work
6. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A. Supplement
A.1. Preparation and Culture of Peripheral Blood Monocytic Cells
A.2. Proliferation Assay
References
- Lassmann, H.; Bruck, W.; Lucchinetti, C.F. The immunopathology of multiple sclerosis: An overview. Brain Pathol. 2007, 17, 210–218. [Google Scholar] [CrossRef] [PubMed]
- Taheri, M.; Ghafouri-Fard, S.; Sayad, A.; Arsang-Jang, S.; Mazdeh, M.; Toghi, M.; Omrani, M.D. Assessment of Protein Prenylation Pathway in Multiple Sclerosis Patients. J. Mol. Neurosci. 2018, 64, 581–590. [Google Scholar] [CrossRef] [PubMed]
- Hauser, S.L.; Oksenberg, J.R. The neurobiology of multiple sclerosis: Genes, inflammation, and neurodegeneration. Neuron 2006, 52, 61–76. [Google Scholar] [CrossRef] [PubMed]
- De Jong, R.; Bezemer, A.C.; Zomerdijk, T.P.; van de Pouw-Kraan, T.; Ottenhoff, T.H.; Nibbering, P.H. Selective stimulation of T helper 2 cytokine responses by the anti-psoriasis agent monomethylfumarate. Eur. J. Immunol. 1996, 26, 2067–2074. [Google Scholar] [CrossRef] [PubMed]
- Kronenberg, J.; Pars, K.; Brieskorn, M.; Prajeeth, C.K.; Heckers, S.; Schwenkenbecher, P.; Skripuletz, T.; Pul, R.; Pavlou, A.; Stangel, M. Fumaric Acids Directly Influence Gene Expression of Neuroprotective Factors in Rodent Microglia. Int. J. Mol. Sci. 2019, 20, 325. [Google Scholar] [CrossRef] [PubMed]
- Linker, R.A.; Lee, D.H.; Ryan, S.; van Dam, A.M.; Conrad, R.; Bista, P.; Zeng, W.; Hronowsky, X.; Buko, A.; Chollate, S.; et al. Fumaric acid esters exert neuroprotective effects in neuroinflammation via activation of the Nrf2 antioxidant pathway. Brain A J. Neurol. 2011, 134, 678–692. [Google Scholar] [CrossRef] [Green Version]
- Scannevin, R.H.; Chollate, S.; Jung, M.Y.; Shackett, M.; Patel, H.; Bista, P.; Zeng, W.; Ryan, S.; Yamamoto, M.; Lukashev, M.; et al. Fumarates promote cytoprotection of central nervous system cells against oxidative stress via the nuclear factor (erythroid-derived 2)-like 2 pathway. J. Pharmacol. Exp. Ther. 2012, 341, 274–284. [Google Scholar] [CrossRef]
- Litjens, N.H.; Burggraaf, J.; van Strijen, E.; van Gulpen, C.; Mattie, H.; Schoemaker, R.C.; van Dissel, J.T.; Thio, H.B.; Nibbering, P.H. Pharmacokinetics of oral fumarates in healthy subjects. Br. J. Clin. Pharmacol. 2004, 58, 429–432. [Google Scholar] [CrossRef] [Green Version]
- Litjens, T.; Harland, M.L.; Roberts, M.L.; Barritt, G.J.; Rychkov, G.Y. Fast Ca(2+)-dependent inactivation of the store-operated Ca2+ current (ISOC) in liver cells: A role for calmodulin. J. Physiol. 2004, 558, 85–97. [Google Scholar] [CrossRef]
- Moharregh-Khiabani, D.; Linker, R.A.; Gold, R.; Stangel, M. Fumaric Acid and its esters: An emerging treatment for multiple sclerosis. Curr. Neuropharmacol. 2009, 7, 60–64. [Google Scholar] [CrossRef]
- Gold, R.; Linker, R.A.; Stangel, M. Fumaric acid and its esters: An emerging treatment for multiple sclerosis with antioxidative mechanism of action. Clin. Immunol. 2012, 142, 44–48. [Google Scholar] [CrossRef] [PubMed]
- Ghoreschi, K.; Bruck, J.; Kellerer, C.; Deng, C.; Peng, H.; Rothfuss, O.; Hussain, R.Z.; Gocke, A.R.; Respa, A.; Glocova, I.; et al. Fumarates improve psoriasis and multiple sclerosis by inducing type II dendritic cells. J. Exp. Med. 2011, 208, 2291–2303. [Google Scholar] [CrossRef] [PubMed]
- Williamson, T.P.; Johnson, D.A.; Johnson, J.A. Activation of the Nrf2-ARE pathway by siRNA knockdown of Keap1 reduces oxidative stress and provides partial protection from MPTP-mediated neurotoxicity. Neurotoxicology 2012, 33, 272–279. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Asadullah, K.; Schmid, H.; Friedrich, M.; Randow, F.; Volk, H.D.; Sterry, W.; Docke, W.D. Influence of monomethylfumarate on monocytic cytokine formation-explanation for adverse and therapeutic effects in psoriasis? Arch. Dermatol. Res. 1997, 289, 623–630. [Google Scholar] [CrossRef] [PubMed]
- Weiner, H.L. The challenge of multiple sclerosis: How do we cure a chronic heterogeneous disease? Ann. Neurol. 2009, 65, 239–248. [Google Scholar] [CrossRef] [PubMed]
- Kimelberg, H.K.; Nedergaard, M. Functions of astrocytes and their potential as therapeutic targets. Neurotherapeutics 2010, 7, 338–353. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sidoryk-Wegrzynowicz, M.; Wegrzynowicz, M.; Lee, E.; Bowman, A.B.; Aschner, M. Role of astrocytes in brain function and disease. Toxicol. Pathol. 2011, 39, 115–123. [Google Scholar] [CrossRef] [PubMed]
- Rothstein, J.D.; Dykes-Hoberg, M.; Pardo, C.A.; Bristol, L.A.; Jin, L.; Kuncl, R.W.; Kanai, Y.; Hediger, M.A.; Wang, Y.; Schielke, J.P.; et al. Knockout of glutamate transporters reveals a major role for astroglial transport in excitotoxicity and clearance of glutamate. Neuron 1996, 16, 675–686. [Google Scholar] [CrossRef]
- Ao, L.Y.; Yan, Y.Y.; Zhou, L.; Li, C.Y.; Li, W.T.; Fang, W.R.; Li, Y.M. Immune Cells After Ischemic Stroke Onset: Roles, Migration, and Target Intervention. J. Mol. Neurosci. 2018, 66, 342–355. [Google Scholar] [CrossRef]
- Skripuletz, T.; Hackstette, D.; Bauer, K.; Gudi, V.; Pul, R.; Voss, E.; Berger, K.; Kipp, M.; Baumgartner, W.; Stangel, M. Astrocytes regulate myelin clearance through recruitment of microglia during cuprizone-induced demyelination. Brain A J. Neurol. 2013, 136, 147–167. [Google Scholar] [CrossRef]
- Wilms, H.; Sievers, J.; Rickert, U.; Rostami-Yazdi, M.; Mrowietz, U.; Lucius, R. Dimethylfumarate inhibits microglial and astrocytic inflammation by suppressing the synthesis of nitric oxide, IL-1beta, TNF-alpha and IL-6 in an in-vitro model of brain inflammation. J. Neuroinflamm. 2010, 7, 30. [Google Scholar] [CrossRef] [PubMed]
- Kotsiari, A.; Voss, E.V.; Pul, R.; Skripuletz, T.; Ragancokova, D.; Trebst, C.; Stangel, M. Interferon-beta treatment normalises the inhibitory effect of serum from multiple sclerosis patients on oligodendrocyte progenitor proliferation. Neurosci. Lett. 2010, 485, 107–111. [Google Scholar] [CrossRef] [PubMed]
- Janssen, S.; Schlegel, C.; Gudi, V.; Prajeeth, C.K.; Skripuletz, T.; Trebst, C.; Stangel, M. Effect of FTY720-phosphate on the expression of inflammation-associated molecules in astrocytes in vitro. Mol. Med. Rep. 2015, 12, 6171–6177. [Google Scholar] [CrossRef] [PubMed]
- Prajeeth, C.K.; Kronisch, J.; Khorooshi, R.; Knier, B.; Toft-Hansen, H.; Gudi, V.; Floess, S.; Huehn, J.; Owens, T.; Korn, T.; et al. Effectors of Th1 and Th17 cells act on astrocytes and augment their neuroinflammatory properties. J. Neuroinflamm. 2017, 14, 204. [Google Scholar] [CrossRef] [PubMed]
- Treumer, F.; Zhu, K.; Glaser, R.; Mrowietz, U. Dimethylfumarate is a potent inducer of apoptosis in human T cells. J. Investig. Dermatol. 2003, 121, 1383–1388. [Google Scholar] [CrossRef] [PubMed]
- Moharregh-Khiabani, D.; Blank, A.; Skripuletz, T.; Miller, E.; Kotsiari, A.; Gudi, V.; Stangel, M. Effects of fumaric acids on cuprizone induced central nervous system de- and remyelination in the mouse. PLoS ONE 2010, 5, e11769. [Google Scholar] [CrossRef]
- Spencer, C.M.; Crabtree-Hartman, E.C.; Lehmann-Horn, K.; Cree, B.A.; Zamvil, S.S. Reduction of CD8(+) T lymphocytes in multiple sclerosis patients treated with dimethyl fumarate. Neurol. Neuroimmunol. Neuroinflamm. 2015, 2, e76. [Google Scholar] [CrossRef]
- Gross, C.C.; Schulte-Mecklenbeck, A.; Klinsing, S.; Posevitz-Fejfar, A.; Wiendl, H.; Klotz, L. Dimethyl fumarate treatment alters circulating T helper cell subsets in multiple sclerosis. Neurol. Neuroimmunol. Neuroinflamm. 2016, 3, e183. [Google Scholar] [CrossRef]
- Chung, I.Y.; Benveniste, E.N. Tumor necrosis factor-alpha production by astrocytes. Induction by lipopolysaccharide, IFN-gamma, and IL-1 beta. J. Immunol. 1990, 144, 2999–3007. [Google Scholar]
- Rosciszewski, G.; Cadena, V.; Murta, V.; Lukin, J.; Villarreal, A.; Roger, T.; Ramos, A.J. Toll-Like Receptor 4 (TLR4) and Triggering Receptor Expressed on Myeloid Cells-2 (TREM-2) Activation Balance Astrocyte Polarization into a Proinflammatory Phenotype. Mol. Neurobiol. 2018, 55, 3875–3888. [Google Scholar] [CrossRef]
- Heinrichs, D.E.; Yethon, J.A.; Whitfield, C. Molecular basis for structural diversity in the core regions of the lipopolysaccharides of Escherichia coli and Salmonella enterica. Mol. Microbiol. 1998, 30, 221–232. [Google Scholar] [CrossRef] [PubMed]
- Regen, T.; van Rossum, D.; Scheffel, J.; Kastriti, M.-E.; Revelo, N.H.; Prinz, M.; Brück, W.; Hanisch, U.-K. CD14 and TRIF govern distinct responsiveness and responses in mouse microglial TLR4 challenges by structural variants of LPS. Brain Behav. Immun. 2011, 25, 957–970. [Google Scholar] [CrossRef] [PubMed]
- Lucchinetti, C.; Bruck, W.; Parisi, J.; Scheithauer, B.; Rodriguez, M.; Lassmann, H. Heterogeneity of multiple sclerosis lesions: Implications for the pathogenesis of demyelination. Ann. Neurol. 2000, 47, 707–717. [Google Scholar] [CrossRef]
- Kornek, B.; Storch, M.K.; Weissert, R.; Wallstroem, E.; Stefferl, A.; Olsson, T.; Linington, C.; Schmidbauer, M.; Lassmann, H. Multiple sclerosis and chronic autoimmune encephalomyelitis: A comparative quantitative study of axonal injury in active, inactive, and remyelinated lesions. Am. J. Pathol. 2000, 157, 267–276. [Google Scholar] [CrossRef]
- Trapp, B.D.; Peterson, J.; Ransohoff, R.M.; Rudick, R.; Mork, S.; Bo, L. Axonal transection in the lesions of multiple sclerosis. N. Engl. J. Med. 1998, 338, 278–285. [Google Scholar] [CrossRef] [PubMed]
- Storch, M.K.; Piddlesden, S.; Haltia, M.; Iivanainen, M.; Morgan, P.; Lassmann, H. Multiple sclerosis: In situ evidence for antibody- and complement-mediated demyelination. Ann. Neurol. 1998, 43, 465–471. [Google Scholar] [CrossRef]
- Storch, M.K.; Stefferl, A.; Brehm, U.; Weissert, R.; Wallstrom, E.; Kerschensteiner, M.; Olsson, T.; Linington, C.; Lassmann, H. Autoimmunity to myelin oligodendrocyte glycoprotein in rats mimics the spectrum of multiple sclerosis pathology. Brain Pathol. 1998, 8, 681–694. [Google Scholar] [CrossRef]
- Aboul-Enein, F.; Rauschka, H.; Kornek, B.; Stadelmann, C.; Stefferl, A.; Bruck, W.; Lucchinetti, C.; Schmidbauer, M.; Jellinger, K.; Lassmann, H. Preferential loss of myelin-associated glycoprotein reflects hypoxia-like white matter damage in stroke and inflammatory brain diseases. J. Neuropathol. Exp. Neurol. 2003, 62, 25–33. [Google Scholar] [CrossRef]
- Lassmann, H. Hypoxia-like tissue injury as a component of multiple sclerosis lesions. J. Neurol. Sci. 2003, 206, 187–191. [Google Scholar] [CrossRef]
- Yang, R.; Dunn, J.F. Multiple sclerosis disease progression: Contributions from a hypoxia-inflammation cycle. Mult. Scler. J. 2018. [Google Scholar] [CrossRef]
- Bartels, K.; Grenz, A.; Eltzschig, H.K. Hypoxia and inflammation are two sides of the same coin. Proc. Natl. Acad. Sci. USA 2013, 110, 18351–18352. [Google Scholar] [CrossRef] [Green Version]
- Eltzschig, H.K.; Carmeliet, P. Hypoxia and inflammation. N. Engl. J. Med. 2011, 364, 656–665. [Google Scholar] [CrossRef]
- Cummins, E.P.; Berra, E.; Comerford, K.M.; Ginouves, A.; Fitzgerald, K.T.; Seeballuck, F.; Godson, C.; Nielsen, J.E.; Moynagh, P.; Pouyssegur, J.; et al. Prolyl hydroxylase-1 negatively regulates IkappaB kinase-beta, giving insight into hypoxia-induced NFkappaB activity. Proc. Natl. Acad. Sci. USA 2006, 103, 18154–18159. [Google Scholar] [CrossRef]
- Rostami-Yazdi, M.; Clement, B.; Schmidt, T.J.; Schinor, D.; Mrowietz, U. Detection of metabolites of fumaric acid esters in human urine: Implications for their mode of action. J. Investig. Dermatol. 2009, 129, 231–234. [Google Scholar] [CrossRef]
- Stoof, T.J.; Flier, J.; Sampat, S.; Nieboer, C.; Tensen, C.P.; Boorsma, D.M. The antipsoriatic drug dimethylfumarate strongly suppresses chemokine production in human keratinocytes and peripheral blood mononuclear cells. Br. J. Dermatol. 2001, 144, 1114–1120. [Google Scholar] [CrossRef]
- Kipp, M.; Norkute, A.; Johann, S.; Lorenz, L.; Braun, A.; Hieble, A.; Gingele, S.; Pott, F.; Richter, J.; Beyer, C. Brain-region-specific astroglial responses in vitro after LPS exposure. J. Mol. Neurosci. 2008, 35, 235–243. [Google Scholar] [CrossRef]
- Sawada, M.; Kondo, N.; Suzumura, A.; Marunouchi, T. Production of tumor necrosis factor-alpha by microglia and astrocytes in culture. Brain Res. 1989, 491, 394–397. [Google Scholar] [CrossRef]
- Xiang, Y.; Wei, X.; Chen, L.; Liu, H.; Liu, X.; Wang, T.; Zhang, X. Anti-inflammatory effect of acetylpuerarin on eicosanoid signaling pathway in primary rat astrocytes. J. Mol. Neurosci. 2014, 52, 577–585. [Google Scholar] [CrossRef]
- Lin, S.X.; Lisi, L.; Dello Russo, C.; Polak, P.E.; Sharp, A.; Weinberg, G.; Kalinin, S.; Feinstein, D.L. The anti-inflammatory effects of dimethyl fumarate in astrocytes involve glutathione and haem oxygenase-1. ASN Neuro 2011, 3, AN20100033. [Google Scholar] [CrossRef]
- Brennan, M.S.; Patel, H.; Allaire, N.; Thai, A.; Cullen, P.; Ryan, S.; Lukashev, M.; Bista, P.; Huang, R.; Rhodes, K.J.; et al. Pharmacodynamics of Dimethyl Fumarate Are Tissue Specific and Involve NRF2-Dependent and -Independent Mechanisms. Antioxid. Redox Signal. 2016, 24, 1058–1071. [Google Scholar] [CrossRef]
- Schulze-Topphoff, U.; Varrin-Doyer, M.; Pekarek, K.; Spencer, C.M.; Shetty, A.; Sagan, S.A.; Cree, B.A.; Sobel, R.A.; Wipke, B.T.; Steinman, L.; et al. Dimethyl fumarate treatment induces adaptive and innate immune modulation independent of Nrf2. Proc. Natl. Acad. Sci. USA 2016, 113, 4777–4782. [Google Scholar] [CrossRef] [Green Version]
- Longbrake, E.E.; Ramsbottom, M.J.; Cantoni, C.; Ghezzi, L.; Cross, A.H.; Piccio, L. Dimethyl fumarate selectively reduces memory T cells in multiple sclerosis patients. Mult. Scler. J. 2016, 22, 1060–1070. [Google Scholar] [CrossRef]
- Wu, Q.; Wang, Q.; Mao, G.; Dowling, C.A.; Lundy, S.K.; Mao-Draayer, Y. Dimethyl Fumarate Selectively Reduces Memory T Cells and Shifts the Balance between Th1/Th17 and Th2 in Multiple Sclerosis Patients. J. Immunol. 2017, 198, 3069–3080. [Google Scholar] [CrossRef] [Green Version]
- Chen, H.; Assmann, J.C.; Krenz, A.; Rahman, M.; Grimm, M.; Karsten, C.M.; Kohl, J.; Offermanns, S.; Wettschureck, N.; Schwaninger, M. Hydroxycarboxylic acid receptor 2 mediates dimethyl fumarate’s protective effect in EAE. J. Clin. Investig. 2014, 124, 2188–2192. [Google Scholar] [CrossRef]
- Prajeeth, C.K.; Huehn, J.; Stangel, M. Regulation of neuroinflammatory properties of glial cells by T cell effector molecules. Neural Regen. Res. 2018, 13, 234–236. [Google Scholar] [CrossRef]
- Maier, T.; Guell, M.; Serrano, L. Correlation of mRNA and protein in complex biological samples. FEBS Lett. 2009, 583, 3966–3973. [Google Scholar] [CrossRef] [Green Version]
- De Sousa Abreu, R.; Penalva, L.O.; Marcotte, E.M.; Vogel, C. Global signatures of protein and mRNA expression levels. Mol. Biosyst. 2009, 5, 1512–1526. [Google Scholar] [CrossRef] [Green Version]
- Tarassishin, L.; Suh, H.S.; Lee, S.C. LPS and IL-1 differentially activate mouse and human astrocytes: Role of CD14. Glia 2014, 62, 999–1013. [Google Scholar] [CrossRef] [Green Version]
- Galloway, D.A.; Williams, J.B.; Moore, C.S. Effects of fumarates on inflammatory human astrocyte responses and oligodendrocyte differentiation. Ann. Clin. Transl. Neurol. 2017, 4, 381–391. [Google Scholar] [CrossRef]
Gene | Gene Expression Assay Number |
---|---|
NGF | Rn_01533872_m1 |
BNDF | Rn_00560868_m1 |
GDNF | Rn_00569510_m1 |
PDGFa | Rn_00709363_m1 |
FGF2 | Rn_00570809_m1 |
CNTF | Rn_00755092_m1 |
IL-1β | Rn_00580432_m1 |
IGF-1 | Rn_00710306_m1 |
TNFα | Rn_99999017_m1 |
iNOS | Rn_00561646_m1 |
IL-6 | Rn_01410330_m1 |
HPRT | Rn_01527840_m1 |
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Pars, K.; Gingele, M.; Kronenberg, J.; Prajeeth, C.K.; Skripuletz, T.; Pul, R.; Jacobs, R.; Gudi, V.; Stangel, M. Fumaric Acids Do Not Directly Influence Gene Expression of Neuroprotective Factors in Highly Purified Rodent Astrocytes. Brain Sci. 2019, 9, 241. https://doi.org/10.3390/brainsci9090241
Pars K, Gingele M, Kronenberg J, Prajeeth CK, Skripuletz T, Pul R, Jacobs R, Gudi V, Stangel M. Fumaric Acids Do Not Directly Influence Gene Expression of Neuroprotective Factors in Highly Purified Rodent Astrocytes. Brain Sciences. 2019; 9(9):241. https://doi.org/10.3390/brainsci9090241
Chicago/Turabian StylePars, Kaweh, Marina Gingele, Jessica Kronenberg, Chittappen K Prajeeth, Thomas Skripuletz, Refik Pul, Roland Jacobs, Viktoria Gudi, and Martin Stangel. 2019. "Fumaric Acids Do Not Directly Influence Gene Expression of Neuroprotective Factors in Highly Purified Rodent Astrocytes" Brain Sciences 9, no. 9: 241. https://doi.org/10.3390/brainsci9090241