Induction of tumor necrosis factor alpha (TNF) expression in microglia by the accumulation of a superoxide anion donor in rat cerebral cortex

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Kazuyuki Nakajima Takashi Ishijima Hiroshi Kobayashi


Previously we reported that endotoxin-dependent induction of tumor necrosis factor alpha (TNFexpression in microglia was significantly suppressed by the superoxide anion scavenger N-acetyl cysteine (NAC), and that microglia induced TNF in response to a superoxide anion donor 3-(4-morpholinyl)sydnonimine (SIN-1) in vitro. Those findings strongly suggested that superoxide anion is associated with the induction of TNF in microglia. However, whether TNF is actually induced in microglia in vivo remains to be determined. In the present study, we confirmed the ability of microglia to induce TNF in vitro and examined the effects of SIN-1 on microglial induction of TNF in vivo. The accumulation of SIN-1 solution in rat cerebral cortex led to the induction of TNF on the ipsilateral, but not the contralateral side. The levels of TNF in the ipsilateral cortex peaked at 6-12 h post-accumulation. Immunohistochemical study revealed that anti-TNF antibody-positive cells in the SIN-1-injected region were mainly anti-ionized Ca 2+ binding adapter molecule-1 (Iba-1) antibody-positive, suggesting that microglia are a major cell type for inducing TNF. On the other hand, interleukin 1beta (IL-1) and IL-6 were not detected in the SIN-1-injected cortex. Together, these results indicate that microglia induced TNF in vivo in response to superoxide anion.

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NAKAJIMA, Kazuyuki; ISHIJIMA, Takashi; KOBAYASHI, Hiroshi. Induction of tumor necrosis factor alpha (TNF) expression in microglia by the accumulation of a superoxide anion donor in rat cerebral cortex. Medical Research Archives, [S.l.], v. 7, n. 5, may 2019. ISSN 2375-1924. Available at: <>. Date accessed: 12 july 2020. doi:


1. Selmaj KW, Raine CS. Tumor necrosis factor mediates myelin and oligodendrocyte damage in vitro. Ann Neurol. 1988;23:339-346

2. Venters HD, Dantzer R, Kelley KW. A new concept in neurodegeneration: TNFis a silencer of survival signals. Trends Neurosci. 2000;23:175-180

3. Zassler B, Weis C, Humpel C. Tumor necrosis factor-a triggers cell death of sensitized potassium chloride-stimulated cholinergic neurons. Brain Res Mol Brain Res. 2003;113:78-85

4. Sedel F, Béchade C, Vyas S, Triller A. Macrophage-derived tumor necrosis factor a, an early developmental signal for motoneuron death. J Neurosci. 2004;24:2236-2246

5. Cueva Vargas JL, Osswald IK, Unsain N, Aurousseau MR, Barker PA, Bowie D, Di Polo A. Soluble tumor necrosis factor a promotes retinal ganglion cell death in glaucoma via calcium-permeable AMPA receptor activation. J Neurosci. 2015;35:12088-12102

6. Prajapati P, Sripada L, Singh K, Bhatelia K, Singh R, Singh R. TNF-α regulates miRNA targeting mitochondrial complex-I and induces cell death in dopaminergic cells.
Biochim Biophys Acta. 2015;1852:451-461

7. Nakajima K, Tohyama Y, Kohsaka S, Kurihara T. Protein kinase C requirement in the activation of p38 mitogen-activated protein kinase, which is linked to the induction of tumor necrosis factor  in lipopolysaccharide-stimulated microglia. Neurochem Int. 2004;44:205-214

8. Yoshino Y, Yamamoto S, Kohsaka S, Oshiro S, Nakajima K. Superoxide anion contributes to the induction of tumor necrosis factor alpha (TNFα) through activation of the MKK3/6-p38 MAPK cascade in rat microglia. Brain Res. 2011;1422:1-12

9. Hogg N, Darley-Usmar VM, Wilson MT, Moncada S. Production of hydroxyl radicals from the simultaneous generation of superoxide and nitric oxide. Biochem J. 1992;281:419-424

10. Nakajima K, Graeber MB, Sonoda M, Tohyama Y, Kohsaka S, Kurihara T. In vitro proliferation of axotomized rat facial nucleus-derived activated microglia in an autocrine fashion. J Neurosci Res. 2006;84:348-359

11. Maeda S, Nakajima K, Tohyama Y, Kohsaka S. Characteristic response of astrocytes to plasminogen/plasmin to upregulate transforming growth factor beta 3 (TGFb3) production/secretion through proteinase-activated receptor-1 (PAR-1) and the downstream phosphatidylinositol 3-kinase (PI3K)-Akt/PKB signaling cascade. Brain Res. 2009;1305:1-13

12. Lowry OJ, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265-275

13. Ichimiya T, Yamamoto S, Honda Y, Kikuchi R, Kohsaka S, Nakajima K. Functional down-regulation of axotomized rat facial motoneurons. Brain Res. 2013;1507:35-44

14. Arnett HA, Mason J, Marino M, Suzuki K, Matsushima GK, Ting JP. TNFa promotes proliferation of oligodendrocyte progenitors and remyelination. Nat Neurosci. 2001;4:1116-1122

15. Leibovich SJ, Polverini PJ, Shepard HM, Wiseman DM, Shively V, Nuseir N. Macrophage-induced angiogenesis is mediated by tumour necrosis factor-a. Nature. 1987;329:630-632

16. Barna BP, Estes ML, Jacobs BS, Hudson S, Ransohoff RM. Human astrocytes proliferate in response to tumor necrosis factor a. J Neuroimmunol. 1990;30:239-343

17. Aloisi F, Carè A, Borsellino G, Gallo P, Rosa S, Bassani A, Cabibbo A, Testa U, Levi G, Peschle C. Production of hemolymphopoietic cytokines (IL-6, IL-8, colony-stimulating factors) by normal human astrocytes in response to IL-1b and tumor necrosis factor-a. J Immunol. 1992;149:2358-2366

18. Panek RB, Lee YJ, Itoh-Lindstrom Y, Ting JP, Benveniste EN. Characterization of astrocyte nuclear proteins involved in IFN-g- and TNF-a-mediated class II MHC gene expression. J Immunol. 1994;153:4555-4564

19. Romero LI, Tatro JB, Field JA, Reichlin S. Roles of IL-1 and TNF-a in endotoxin-induced activation of nitric oxide synthase in cultured rat brain cells. Am J Physiol. 1996;270:R326-332

20. Schwartz M, Solomon A, Lavie V, Ben-Bassat S, Belkin M, Cohen A. Tumor necrosis factor facilitates regeneration of injured central nervous system axons. Brain Res. 1991;545:334-338

21. Neumann H, Schweigreiter R, Yamashita T, Rosenkranz K, Wekerle H, Barde YA. Tumor necrosis factor inhibits neurite outgrowth and branching of hippocampal neurons by a rho-dependent mechanism. J Neurosci. 2002;22:854-862

22. von Zahn J, Möller T, Kettenmann H, Nolte C. Microglial phagocytosis is modulated by pro- and anti-inflammatory cytokines. Neuroreport. 1997;8:3851-3856

23. Buttini M, Appel K, Sauter A, Gebicke-Haerter PJ, Boddeke HW. Expression of tumor necrosis factor  after focal cerebral ischaemia in the rat. Neurosci. 1996;71:1-16

24. Medana IM, Hunt NH, Chaudhri G. Tumor necrosis factor- expression in the brain during fatal murine cerebral malaria: evidence for production by microglia and astrocytes. Am J Pathol. 1997;150:1473-1486

25. Uno H, Matsuyama T, Akita H, Nishimura H, Sugita M. Induction of tumor necrosis factor-a in the mouse hippocampus following transient forebrain ischemia. J Cereb Blood Flow Metab. 1997;17:491-499

26. Barone FC, Arvin B, White RF, Miller A, Webb CL, Willette RN, Lysko PG, Feuerstein GZ. Tumor necrosis factor-. A mediator of focal ischemic brain injury. Stroke 1997;28:1233-1244

27. Maccioni RB, Rojo LE, Fernández JA, Kuljis RO. The role of neuroimmunomodulation in Alzheimer's disease. Ann NY Acad Sci. 2009;1153:240-246

28. Reale M, Iarlori C, Thomas A, Gambi D, Perfetti B, Di Nicola M, Onofrj M. Peripheral cytokines profile in Parkinson's disease. Brain Behav Immun. 2009;23:55-63

29. Cannella B, Raine CS. The adhesion molecule and cytokine profile of multiple sclerosis lesions. Ann Neurol. 1995;37:424-435

30. Yeung MC, Pulliam L, Lau AS. The HIV envelope protein gp120 is toxic to human brain-cell cultures through the induction of interleukin-6 and tumor necrosis factor-. AIDS. 1995;9:137-143

31. Banati RB, Gehrmann J, Schubert P, Kreutzberg GW. Cytotoxicity of microglia. Glia. 1993;7:111-118

32. Benveniste EN. Role of macrophages/microglia in multiple sclerosis and experimental allergic encephalomyelitis. J Mol Med (Berl). 1997;75:165-173

33. Gregersen R, Lambertsen K, Finsen B. Microglia and macrophages are the major source of tumor necrosis factor in permanent middle cerebral artery occlusion in mice. J Cereb Blood Flow Metab. 2000;20:53-65

34. Kempuraj D, Thangavel R, Natteru PA, Selvakumar GP, Saeed D, Zahoor H, Zaheer S, Iyer SS, Zaheer A. Neuroinflammation induces neurodegeneration. J Neurol Neurosurg Spine. 2016;1(1) 1-15

35. Cacci E, Claasen JH, Kokaia Z. Microglia-derived tumor necrosis factor-a exaggerates death of newborn hippocampal progenitor cells in vitro. J Neurosci Res. 2005;80:789-797

36. Chung IY, Benveniste EN. Tumor necrosis factor-a production by astrocytes. Induction by lipopolysaccharide, IFN-g, and IL-1b. J Immunol. 1990;144:2999-3007

37. Kim HM, Shin HY, Lim KH, Ryu ST, Shin TY, Chae HJ, Kim HR, Lyu YS, An NH, Lim KS. Taraxacum officinale inhibits tumor necrosis factor-a production from rat astrocytes. Immunopharmacol Immunotoxicol. 2000;22:519-530

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