Effect of microglia on iron metabolism in midbrain dopaminergic neurons and the underlying mechanism: study protocol for an
in vitro cellular experiment

doi:10.3969/j.issn.2095-4344.2017.08.020

Abstract

Abstract:

BACKGROUND: Iron metabolism disorder has been proved to be a key factor of Parkinson’s disease, but research on brain iron metabolism is still immature, and the role of glial cells in the brain iron metabolism remains unclear. Many studies have pointed that microglia plays an important role in the pathogenesis of Parkinson’s disease, which closely related to iron.
OBJECTIVE: To further explore the relationship between microglia and neurons in iron metabolism based on our previous study, and to clarify the role of microglia in iron-induced selective damaged dopaminergic neurons of Parkinson’s disease.
METHODS: This was an in vitro cellular experiment, which was finished in the Medical College of Qingdao University, Shandong Province, China. The effects of conditioned medium of microglia on the survival rate of mesencephalic neurons and iron metabolism were observed by changing the iron levels in microglia. Lipopolysaccharide was used to activate different iron-loaded microglia, and then the effects of conditioned medium on the survival rate of mesencephalic neurons and iron metabolism were determined. The effects of different iron levels in microglia on microglia and inflammatory factors and lactoferrin released from the activated microglia were detected by siRNA and ELISA. The effects of these inflammatory factors and lactoferrin on the survival of mesencephalic neurons and iron metabolism were observed. The iron levels in mesencephalic neurons were changed, and then added into the above conditioned medium to observe the survival of neurons and iron metabolism. This experimental protocol was approved by the Hospital Ethics Committee (No. 01482 311234). All experimental procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals issued by the United States National Institutes of Health.
RESULTS AND CONCLUSION: Our study results show that the function of microglia is closely related to the iron metabolism in nigra dopaminergic neurons. Research on the rational use of microglia to prevent neuronal degeneration gives a strong impetus to the prevention and treatment of Parkinson’s disease.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Microglia, Parkinson Disease, Iron, Tissue Engineering

CLC Number:

R318

Shi Cheng-kui, Chen Zhi-qiang. Effect of microglia on iron metabolism in midbrain dopaminergic neurons and the underlying mechanism: study protocol for an
in vitro cellular experiment[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(8): 1262-1267.

Figures/Tables 2

References

[1] Mirza B, Hadberg H, Thomsen P, et al. The absence of reactive astrocytosis is indicative of a unique inflammatory process in Parkinson’s disease. Neuroscience. 2000;95: 425-432.

[2] Zhang X, Surguladze N, Slagle-Webb B, et al. Cellular iron status influences the functional relationship between microglia and oligodendrocytes. Glia. 2006;54(8):795-804.

[3] Wu DC, Jackson-Lewis V, Vila M, et al. Blockade of microglial activation is neuroprotective in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine mouse model of Parkinson disease. J Neurosci. 2002;22(5):1763-1771.

[4] Peng J, Stevenson FF, Oo ML, et al. Iron-enhanced paraquat-mediated dopaminergic cell death due to increased oxidative stress as a consequence of microglial activation. Free Radic Biol Med. 2009;46(2):312-320.

[5] O'Brien-Ladner AR, Nelson SR, Murphy WJ, et al. Iron is a regulatory component of human IL-1beta production. Support for regional variability in the lung. Am J Respir Cell Mol Biol. 2000;23(1):112-119.

[6] Helyar L, Sherman AR. Iron deficiency and interleukin 1 production by rat leukocytes. Am J Clin Nutr. 1987;46(2): 346-352.

[7] Smirnov IM, Bailey K, Flowers CH, et al. Effects of TNF-alpha and IL-1beta on iron metabolism by A549 cells and influence on cytotoxicity. Am J Physiol. 1999;277(2 Pt1):L257-263.

[8] Eisenstein RS, Tuazon PT, Schalinske KL, et al. Iron-responsive element-binding protein. Phosphorylation by protein kinase C. J Biol Chem. 1993;268:27363-27370.

[9] Schalinske KL, Eisenstein RS. Phosphorylation and activation of both iron regulatory proteins 1 and 2 in HL-60 cells. J Biol Chem. 1996;271:7168-7176.

[10] Kiemer AK, Förnges AC, Pantopoulos K, et al. ANP-induced decrease of iron regulatory protein activity is independent of HO-1 induction. Am J Physiol Gastrointest Liver Physiol. 2004;287:G518-6526.

[11] An L, Sato H, Konishi Y, et al. Expression and localization of lactotransferrin messenger RNA in the cortex of Alzheimer's disease. Neurosci Lett. 2009;452(3):277-280.

[12] Leveugle B, Faucheux BA, Bouras C, et al. Cellular distribution of the iron-binding protein lactotransferrin in the mesencephalon of Parkinson's disease cases. Acta Neuropathol. 1996;91(6):566-572.

[13] Wang J, Jiang H, Xie JX. Time dependent effects of 6-OHDA lesions on iron level and neuronal loss in rat nigrostriatal system. Neurochem Res. 2004;29(12): 2239-2243.

[14] Zhang S, Wang J, Song N, et al. Up-regulation of divalent metal transporter1 is involved in 1-methyl-4-phenylpyridinium (MPP(+))-induced apoptosis in MES23.5 cells. Neurobiol Aging. 2009;30(9):1466-1476.

[15] Wang J, Jiang H, Xie JX. Ferroportin1 and hephaestin are involved in the nigral iron accumulation of 6-OHDA- lesioned rats. Eur J Neurosci. 2007;25(9):2766-2772.

[16] Xu HM, Jiang H, Wang J, et al. Over-expressed human divalent metal transporter 1 is involved in iron accumulation in MES23.5 cells. Neurochem Int. 2008;52(6):1044-1051.

[17] Song N, Wang J, Jiang H, et al. Ferroportin 1 but not hephaestin contributes to iron accumulation in a cell model of Parkinson's disease. Free Radic Biol Med. 2010;48(2): 332-341.

[18] Youdim MB, Fridkin M, Zheng H. Bifunctional drug derivatives of MAO-B inhibitor rasagiline and iron chelator VK-28 as a more effective approach to treatment of brain ageing and ageing neurodegenerative diseases. Mech Ageing Dev. 2005;126(2):317-326.

[19] Hirsch EC. Altered regulation of iron transport and storage in Parkinson's disease. J Neural Transm Suppl. 2006;(71): 201-204.