Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
Journal of Neuroinflammation
Published in: Journal of Neuroinflammation 1/2011

Open Access 01-12-2011 | Research

Pro-inflammatory cytokines and lipopolysaccharide induce changes in cell morphology, and upregulation of ERK1/2, iNOS and sPLA2-IIA expression in astrocytes and microglia

Authors: Wenwen Sheng, Yijia Zong, Arwa Mohammad, Deepa Ajit, Jiankun Cui, Dongdong Han, Jennifer L Hamilton, Agnes Simonyi, Albert Y Sun, Zezong Gu, Jau-Shyong Hong, Gary A Weisman, Grace Y Sun

Published in: Journal of Neuroinflammation | Issue 1/2011

Login to get access

Abstract

Background

Activation of glial cells, including astrocytes and microglia, has been implicated in the inflammatory responses underlying brain injury and neurodegenerative diseases including Alzheimer's and Parkinson's diseases. Although cultured astrocytes and microglia are capable of responding to pro-inflammatory cytokines and lipopolysaccharide (LPS) in the induction and release of inflammatory factors, no detailed analysis has been carried out to compare the induction of iNOS and sPLA2-IIA. In this study, we investigated the effects of cytokines (TNF-alpha, IL-1beta, and IFN-gamma) and LPS + IFN-gamma to induce temporal changes in cell morphology and induction of p-ERK1/2, iNOS and sPLA2-IIA expression in immortalized rat (HAPI) and mouse (BV-2) microglial cells, immortalized rat astrocytes (DITNC), and primary microglia and astrocytes.

Methods/Results

Cytokines (TNF-alpha, IL-1beta, and IFN-gamma) and LPS + IFN-gamma induced a time-dependent increase in fine processes (filopodia) in microglial cells but not in astrocytes. Filopodia production was attributed to IFN-gamma and was dependent on ERK1/2 activation. Cytokines induced an early (15 min) and a delayed phase (1 ~ 4 h) increase in p-ERK1/2 expression in microglial cells, and the delayed phase increase corresponded to the increase in filopodia production. In general, microglial cells are more active in responding to cytokines and LPS than astrocytes in the induction of NO. Although IFN-gamma and LPS could individually induce NO, additive production was observed when IFN-gamma was added together with LPS. On the other hand, while TNF-alpha, IL-1beta, and LPS could individually induce sPLA2-IIA mRNA and protein expression, this induction process does not require IFN-gamma. Interestingly, neither rat immortalized nor primary microglial cells were capable of responding to cytokines and LPS in the induction of sPLA2-IIA expression.

Conclusion

These results demonstrated the utility of BV-2 and HAPI cells as models for investigation on cytokine and LPS induction of iNOS, and DITNC astrocytes for induction of sPLA2-IIA. In addition, results further demonstrated that cytokine-induced sPLA2-IIA is attributed mainly to astrocytes and not microglial cells.
Appendix
Available only for authorised users
Literature
1.
Benveniste EN, Nguyen VT, O'Keefe GM: Immunological aspects of microglia: relevance to Alzheimer's disease. Neurochem Int. 2001, 39: 381-391. 10.1016/S0197-0186(01)00045-6.CrossRefPubMed
2.
Yenari MA, Kauppinen TM, Swanson RA: Microglial activation in stroke: therapeutic targets. Neurotherapeutics. 2011, 7: 378-391.CrossRef
3.
Van Eldik LJ, Thompson WL, Ralay Ranaivo H, Behanna HA, Martin Watterson D: Glia proinflammatory cytokine upregulation as a therapeutic target for neurodegenerative diseases: function-based and target-based discovery approaches. Int Rev Neurobiol. 2007, 82: 277-296.CrossRefPubMed
4.
5.
6.
Schubert P, Ferroni S: Pathological glial reactions in neurodegenerative disorders: prospects for future therapeutics. Expert Rev Neurother. 2003, 3: 279-287. 10.1586/14737175.3.3.279.CrossRefPubMed
7.
Lu X, Ma L, Ruan L, Kong Y, Mou H, Zhang Z, Wang Z, Wang JM, Le Y: Resveratrol differentially modulates inflammatory responses of microglia and astrocytes. J Neuroinflammation. 2010, 7: 46-10.1186/1742-2094-7-46.PubMedCentralCrossRefPubMed
8.
Possel H, Noack H, Putzke J, Wolf G, Sies H: Selective upregulation of inducible nitric oxide synthase (iNOS) by lipopolysaccharide (LPS) and cytokines in microglia: in vitro and in vivo studies. Glia. 2000, 32: 51-59. 10.1002/1098-1136(200010)32:1<51::AID-GLIA50>3.0.CO;2-4.CrossRefPubMed
9.
Shen S, Yu S, Binek J, Chalimoniuk M, Zhang X, Lo SC, Hannink M, Wu J, Fritsche K, Donato R, Sun GY: Distinct signaling pathways for induction of type II NOS by IFNgamma and LPS in BV-2 microglial cells. Neurochem Int. 2005, 47: 298-307. 10.1016/j.neuint.2005.03.007.CrossRefPubMed
10.
Thampithak A, Jaisin Y, Meesarapee B, Chongthammakun S, Piyachaturawat P, Govitrapong P, Supavilai P, Sanvarinda Y: Transcriptional regulation of iNOS and COX-2 by a novel compound from Curcuma comosa in lipopolysaccharide-induced microglial activation. Neurosci Lett. 2009, 462: 171-175. 10.1016/j.neulet.2009.06.094.CrossRefPubMed
11.
Jana M, Anderson JA, Saha RN, Liu X, Pahan K: Regulation of inducible nitric oxide synthase in proinflammatory cytokine-stimulated human primary astrocytes. Free Radic Biol Med. 2005, 38: 655-664. 10.1016/j.freeradbiomed.2004.11.021.CrossRefPubMed
12.
Jung HW, Chung YS, Kim YS, Park YK: Celastrol inhibits production of nitric oxide and proinflammatory cytokines through MAPK signal transduction and NF-kappaB in LPS-stimulated BV-2 microglial cells. Exp Mol Med. 2007, 39: 715-721.CrossRefPubMed
13.
Murakami M, Taketomi Y, Girard C, Yamamoto K, Lambeau G: Emerging roles of secreted phospholipase A2 enzymes: Lessons from transgenic and knockout mice. Biochimie. 2010, 92: 561-582. 10.1016/j.biochi.2010.03.015.CrossRefPubMed
14.
Ibeas E, Fuentes L, Martin R, Hernandez M, Nieto ML: Secreted phospholipase A2 type IIA as a mediator connecting innate and adaptive immunity: new role in atherosclerosis. Cardiovasc Res. 2009, 81: 54-63.CrossRefPubMed
15.
Kimura-Matsumoto M, Ishikawa Y, Komiyama K, Tsuruta T, Murakami M, Masuda S, Akasaka Y, Ito K, Ishiguro S, Morita H, et al: Expression of secretory phospholipase A2s in human atherosclerosis development. Atherosclerosis. 2008, 196: 81-91. 10.1016/j.atherosclerosis.2006.08.062.CrossRefPubMed
16.
Lin TN, Wang Q, Simonyi A, Chen JJ, Cheung WM, He YY, Xu J, Sun AY, Hsu CY, Sun GY: Induction of secretory phospholipase A2 in reactive astrocytes in response to transient focal cerebral ischemia in the rat brain. J Neurochem. 2004, 90: 637-645. 10.1111/j.1471-4159.2004.02540.x.CrossRefPubMed
17.
Adibhatla RM, Hatcher JF: Secretory phospholipase A2 IIA is up-regulated by TNF-alpha and IL-1alpha/beta after transient focal cerebral ischemia in rat. Brain Res. 2007, 1134: 199-205.PubMedCentralCrossRefPubMed
18.
Moses GS, Jensen MD, Lue LF, Walker DG, Sun AY, Simonyi A, Sun GY: Secretory PLA2-IIA: a new inflammatory factor for Alzheimer's disease. J Neuroinflammation. 2006, 3: 28-10.1186/1742-2094-3-28.PubMedCentralCrossRefPubMed
19.
Liu NK, Zhang YP, Titsworth WL, Jiang X, Han S, Lu PH, Shields CB, Xu XM: A novel role of phospholipase A2 in mediating spinal cord secondary injury. Ann Neurol. 2006, 59: 606-619. 10.1002/ana.20798.CrossRefPubMed
20.
Titsworth WL, Cheng X, Ke Y, Deng L, Burckardt KA, Pendleton C, Liu NK, Shao H, Cao QL, Xu XM: Differential expression of sPLA2 following spinal cord injury and a functional role for sPLA2-IIA in mediating oligodendrocyte death. Glia. 2009, 57: 1521-1537. 10.1002/glia.20867.PubMedCentralCrossRefPubMed
21.
Jensen MD, Sheng W, Simonyi A, Johnson GS, Sun AY, Sun GY: Involvement of oxidative pathways in cytokine-induced secretory phospholipase A2-IIA in astrocytes. Neurochem Int. 2009, 55: 362-368. 10.1016/j.neuint.2009.04.002.PubMedCentralCrossRefPubMed
22.
Li W, Xia J, Sun GY: Cytokine induction of iNOS and sPLA2 in immortalized astrocytes (DITNC): response to genistein and pyrrolidine dithiocarbamate. J Interferon Cytokine Res. 1999, 19: 121-127. 10.1089/107999099314261.CrossRefPubMed
23.
Oka S, Arita H: Inflammatory factors stimulate expression of group II phospholipase A2 in rat cultured astrocytes. Two distinct pathways of the gene expression. J Biol Chem. 1991, 266: 9956-9960.PubMed
24.
Kennedy BP, Payette P, Mudgett J, Vadas P, Pruzanski W, Kwan M, Tang C, Rancourt DE, Cromlish WA: A natural disruption of the secretory group II phospholipase A2 gene in inbred mouse strains. J Biol Chem. 1995, 270: 22378-22385. 10.1074/jbc.270.38.22378.CrossRefPubMed
25.
Cheepsunthorn P, Radov L, Menzies S, Reid J, Connor JR: Characterization of a novel brain-derived microglial cell line isolated from neonatal rat brain. Glia. 2001, 35: 53-62. 10.1002/glia.1070.CrossRefPubMed
26.
Horvath RJ, Nutile-McMenemy N, Alkaitis MS, Deleo JA: Differential migration, LPS-induced cytokine, chemokine, and NO expression in immortalized BV-2 and HAPI cell lines and primary microglial cultures. J Neurochem. 2008, 107: 557-569. 10.1111/j.1471-4159.2008.05633.x.PubMedCentralCrossRefPubMed
27.
McCarthy KD, de Vellis J: Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol. 1980, 85: 890-902. 10.1083/jcb.85.3.890.CrossRefPubMed
28.
Xu J, Chalimoniuk M, Shu Y, Simonyi A, Sun AY, Gonzalez FA, Weisman GA, Wood WG, Sun GY: Prostaglandin E2 production in astrocytes: regulation by cytokines, extracellular ATP, and oxidative agents. Prostaglandins Leukot Essent Fatty Acids. 2003, 69: 437-448. 10.1016/j.plefa.2003.08.016.CrossRefPubMed
29.
Li N, McLaren JE, Michael DR, Clement M, Fielding CA, Ramji DP: ERK is integral to the IFN-gamma-mediated activation of STAT1, the expression of key genes implicated in atherosclerosis, and the uptake of modified lipoproteins by human macrophages. J Immunol. 2010, 185: 3041-3048. 10.4049/jimmunol.1000993.CrossRefPubMed
30.
Chim SM, Qin A, Tickner J, Pavlos N, Davey T, Wang H, Guo Y, Zheng MH, Xu J: EGFL6 Promotes Endothelial Cell Migration and Angiogenesis through the Activation of Extracellular Signal-regulated Kinase. J Biol Chem. 286: 22035-22046.
31.
Nakamura Y, Si QS, Kataoka K: Lipopolysaccharide-induced microglial activation in culture: temporal profiles of morphological change and release of cytokines and nitric oxide. Neurosci Res. 1999, 35: 95-100. 10.1016/S0168-0102(99)00071-1.CrossRefPubMed
32.
Chu SC, Marks-Konczalik J, Wu HP, Banks TC, Moss J: Analysis of the cytokine-stimulated human inducible nitric oxide synthase (iNOS) gene: characterization of differences between human and mouse iNOS promoters. Biochem Biophys Res Commun. 1998, 248: 871-878. 10.1006/bbrc.1998.9062.CrossRefPubMed
33.
Jung JS, Kim DH, Kim HS: Ginsenoside Rh1 suppresses inducible nitric oxide synthase gene expression in IFN-gamma-stimulated microglia via modulation of JAK/STAT and ERK signaling pathways. Biochem Biophys Res Commun. 2010, 397: 323-328. 10.1016/j.bbrc.2010.05.117.CrossRefPubMed
34.
Kozuka N, Itofusa R, Kudo Y, Morita M: Lipopolysaccharide and proinflammatory cytokines require different astrocyte states to induce nitric oxide production. J Neurosci Res. 2005, 82: 717-728. 10.1002/jnr.20671.CrossRefPubMed
35.
36.
Wang JH, Sun GY: Platelet activating factor (PAF) antagonists on cytokine induction of iNOS and sPLA2 in immortalized astrocytes (DITNC). Neurochem Res. 2000, 25: 613-619. 10.1023/A:1007550801444.CrossRefPubMed
Metadata
Title
Pro-inflammatory cytokines and lipopolysaccharide induce changes in cell morphology, and upregulation of ERK1/2, iNOS and sPLA2-IIA expression in astrocytes and microglia
Authors
Wenwen Sheng
Yijia Zong
Arwa Mohammad
Deepa Ajit
Jiankun Cui
Dongdong Han
Jennifer L Hamilton
Agnes Simonyi
Albert Y Sun
Zezong Gu
Jau-Shyong Hong
Gary A Weisman
Grace Y Sun
Publication date
01-12-2011
Publisher
BioMed Central
Published in
Journal of Neuroinflammation / Issue 1/2011
Electronic ISSN: 1742-2094
DOI
https://doi.org/10.1186/1742-2094-8-121

Other articles of this Issue 1/2011

Journal of Neuroinflammation 1/2011 Go to the issue

Research

Granulocyte colony stimulating factor attenuates inflammation in a mouse model of amyotrophic lateral sclerosis

Research

Increased expression of cystine/glutamate antiporter in multiple sclerosis

Research

The formyl peptide receptor like-1 and scavenger receptor MARCO are involved in glial cell activation in bacterial meningitis

Research

CD200-CD200R dysfunction exacerbates microglial activation and dopaminergic neurodegeneration in a rat model of Parkinson's disease

Research

Overexpression of serine racemase in retina and overproduction of D-serine in eyes of streptozotocin-induced diabetic retinopathy

Research

Insulin-like growth factor-I gene delivery to astrocytes reduces their inflammatory response to lipopolysaccharide