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Published in: Journal of Neuroinflammation 1/2015

Open Access 01-12-2015 | Research

Scutellarin regulates the Notch pathway and affects the migration and morphological transformation of activated microglia in experimentally induced cerebral ischemia in rats and in activated BV-2 microglia

Authors: Yun Yuan, Parakalan Rangarajan, Enci Mary Kan, Yajun Wu, Chunyun Wu, Eng-Ang Ling

Published in: Journal of Neuroinflammation | Issue 1/2015

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Abstract

Background

Activated microglial cells release an excess of inflammatory mediators after an ischemic stroke. We reported previously that scutellarin effectively suppressed the inflammatory response induced by activated microglia in rats subjected to middle cerebral artery occlusion (MCAO); however, the mechanism via which scutellarin exerts its effects on microglial activation has not been explored. This study aimed to elucidate if scutellarin can regulate the Notch pathway that is linked to microglia activation in MCAO rat, and in lipopolysaccharide (LPS)-induced BV-2 microglia. Along with this, we also investigated some characteristic behavioral responses of activated microglia.

Methods

Expression of various members of the Notch pathway, including Notch-1, intracellular Notch receptor domain (NICD), recombining binding protein suppressor of hairless (RBP-JK) and transcription factor hairy and enhancer of split-1 (Hes-1) in activated microglia was assessed by immunofluorescence staining and western blot after experimental MCAO and in vitro LPS activation. The effect of scutellarin on migration of microglia was determined by the transwell chamber assay as well as expression of monocyte chemoattractant protein-1 (MCP-1). The morphological change of microglia induced by scutellarin was detected by F-actin staining and electron microscopy.

Results

Scutellarin markedly attenuated the expression of NF-κB, Notch-1, NICD, RBP-JK and Hes-1 both in vivo and in activated microglia. It decreased the expression of MCP-1 and microglial migration, but increased the ability of microglia adhesion. Scutellarin also altered the phenotype of microglia by causing rearrangement or reorganization of its cytoskeleton.

Conclusions

The results suggest that scutellarin regulates the activation of microglia via the Notch pathway and concurrently induces morphological and functional changes in activated microglia.
Appendix
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Literature
2.
4.
go back to reference Gerhard A, Neumaier B, Elitok E, Glatting G, Ries V, Tomczak R, et al. In vivo imaging of activated microglia using [11C]PK11195 and positron emission tomography in patients after ischemic stroke. Neuroreport. 2000;11:2957–60.PubMedCrossRef Gerhard A, Neumaier B, Elitok E, Glatting G, Ries V, Tomczak R, et al. In vivo imaging of activated microglia using [11C]PK11195 and positron emission tomography in patients after ischemic stroke. Neuroreport. 2000;11:2957–60.PubMedCrossRef
5.
go back to reference Neher JJ, Emmrich JV, Fricker M, Mander PK, Thery C, Brown GC. Phagocytosis executes delayed neuronal death after focal brain ischemia. Proc Natl Acad Sci U S A. 2013;110:E4098–107.PubMedCentralPubMedCrossRef Neher JJ, Emmrich JV, Fricker M, Mander PK, Thery C, Brown GC. Phagocytosis executes delayed neuronal death after focal brain ischemia. Proc Natl Acad Sci U S A. 2013;110:E4098–107.PubMedCentralPubMedCrossRef
6.
go back to reference Wang JY, Gualco E, Peruzzi F, Sawaya BE, Passiatore G, Marcinkiewicz C, et al. Interaction between serine phosphorylated IRS-1 and beta1-integrin affects the stability of neuronal processes. J Neurosci Res. 2007;85:2360–73.PubMedCentralPubMedCrossRef Wang JY, Gualco E, Peruzzi F, Sawaya BE, Passiatore G, Marcinkiewicz C, et al. Interaction between serine phosphorylated IRS-1 and beta1-integrin affects the stability of neuronal processes. J Neurosci Res. 2007;85:2360–73.PubMedCentralPubMedCrossRef
7.
go back to reference Lalancette-Hebert M, Phaneuf D, Soucy G, Weng YC, Kriz J. Live imaging of Toll-like receptor 2 response in cerebral ischaemia reveals a role of olfactory bulb microglia as modulators of inflammation. Brain. 2009;132:940–54.PubMedCrossRef Lalancette-Hebert M, Phaneuf D, Soucy G, Weng YC, Kriz J. Live imaging of Toll-like receptor 2 response in cerebral ischaemia reveals a role of olfactory bulb microglia as modulators of inflammation. Brain. 2009;132:940–54.PubMedCrossRef
8.
go back to reference Hong H, Liu GQ. Protection against hydrogen peroxide-induced cytotoxicity in PC12 cells by scutellarin. Life Sci. 2004;74:2959–73.PubMedCrossRef Hong H, Liu GQ. Protection against hydrogen peroxide-induced cytotoxicity in PC12 cells by scutellarin. Life Sci. 2004;74:2959–73.PubMedCrossRef
9.
go back to reference Liu H, Yang X, Tang R, Liu J, Xu H. Effect of scutellarin on nitric oxide production in early stages of neuron damage induced by hydrogen peroxide. Pharmacol Res. 2005;51:205–10.PubMedCrossRef Liu H, Yang X, Tang R, Liu J, Xu H. Effect of scutellarin on nitric oxide production in early stages of neuron damage induced by hydrogen peroxide. Pharmacol Res. 2005;51:205–10.PubMedCrossRef
10.
go back to reference Wang S, Wang H, Guo H, Kang L, Gao X, Hu L. Neuroprotection of Scutellarin is mediated by inhibition of microglial inflammatory activation. Neuroscience. 2011;185:150–60.PubMedCrossRef Wang S, Wang H, Guo H, Kang L, Gao X, Hu L. Neuroprotection of Scutellarin is mediated by inhibition of microglial inflammatory activation. Neuroscience. 2011;185:150–60.PubMedCrossRef
11.
go back to reference Lin LL, Liu AJ, Liu JG, Yu XH, Qin LP, Su DF. Protective effects of scutellarin and breviscapine on brain and heart ischemia in rats. J Cardiovasc Pharmacol. 2007;50:327–32.PubMedCrossRef Lin LL, Liu AJ, Liu JG, Yu XH, Qin LP, Su DF. Protective effects of scutellarin and breviscapine on brain and heart ischemia in rats. J Cardiovasc Pharmacol. 2007;50:327–32.PubMedCrossRef
12.
go back to reference Lutolf S, Radtke F, Aguet M, Suter U, Taylor V. Notch1 is required for neuronal and glial differentiation in the cerebellum. Development. 2002;129:373–85.PubMed Lutolf S, Radtke F, Aguet M, Suter U, Taylor V. Notch1 is required for neuronal and glial differentiation in the cerebellum. Development. 2002;129:373–85.PubMed
13.
go back to reference Alberi L, Hoey SE, Brai E, Scotti AL, Marathe S. Notch signaling in the brain: in good and bad times. Ageing Res Rev. 2013;12:801–14.PubMedCrossRef Alberi L, Hoey SE, Brai E, Scotti AL, Marathe S. Notch signaling in the brain: in good and bad times. Ageing Res Rev. 2013;12:801–14.PubMedCrossRef
14.
go back to reference Cao Q, Lu J, Kaur C, Sivakumar V, Li F, Cheah PS, et al. Expression of Notch-1 receptor and its ligands Jagged-1 and Delta-1 in amoeboid microglia in postnatal rat brain and murine BV-2 cells. Glia. 2008;56:1224–37.PubMedCrossRef Cao Q, Lu J, Kaur C, Sivakumar V, Li F, Cheah PS, et al. Expression of Notch-1 receptor and its ligands Jagged-1 and Delta-1 in amoeboid microglia in postnatal rat brain and murine BV-2 cells. Glia. 2008;56:1224–37.PubMedCrossRef
15.
go back to reference Yao L, Kan EM, Kaur C, Dheen ST, Hao A, Lu J, et al. Notch-1 signaling regulates microglia activation via NF-kappaB pathway after hypoxic exposure in vivo and in vitro. PLoS ONE. 2013;8:e78439.PubMedCentralPubMedCrossRef Yao L, Kan EM, Kaur C, Dheen ST, Hao A, Lu J, et al. Notch-1 signaling regulates microglia activation via NF-kappaB pathway after hypoxic exposure in vivo and in vitro. PLoS ONE. 2013;8:e78439.PubMedCentralPubMedCrossRef
16.
go back to reference Xie J, Wang W, Si JW, Miao XY, Li JC, Wang YC, et al. Notch signaling regulates CXCR4 expression and the migration of mesenchymal stem cells. Cell Immunol. 2013;281:68–75.PubMedCrossRef Xie J, Wang W, Si JW, Miao XY, Li JC, Wang YC, et al. Notch signaling regulates CXCR4 expression and the migration of mesenchymal stem cells. Cell Immunol. 2013;281:68–75.PubMedCrossRef
17.
go back to reference Bolos V, Mira E, Martinez-Poveda B, Luxan G, Canamero M, Martinez AC, et al. Notch activation stimulates migration of breast cancer cells and promotes tumor growth. Breast Cancer Res. 2013;15:R54.PubMedCentralPubMedCrossRef Bolos V, Mira E, Martinez-Poveda B, Luxan G, Canamero M, Martinez AC, et al. Notch activation stimulates migration of breast cancer cells and promotes tumor growth. Breast Cancer Res. 2013;15:R54.PubMedCentralPubMedCrossRef
18.
go back to reference Guentchev M, McKay RD. Notch controls proliferation and differentiation of stem cells in a dose-dependent manner. Eur J Neurosci. 2006;23:2289–96.PubMedCrossRef Guentchev M, McKay RD. Notch controls proliferation and differentiation of stem cells in a dose-dependent manner. Eur J Neurosci. 2006;23:2289–96.PubMedCrossRef
19.
go back to reference VanDussen KL, Carulli AJ, Keeley TM, Patel SR, Puthoff BJ, Magness ST, et al. Notch signaling modulates proliferation and differentiation of intestinal crypt base columnar stem cells. Development. 2012;139:488–97.PubMedCentralPubMedCrossRef VanDussen KL, Carulli AJ, Keeley TM, Patel SR, Puthoff BJ, Magness ST, et al. Notch signaling modulates proliferation and differentiation of intestinal crypt base columnar stem cells. Development. 2012;139:488–97.PubMedCentralPubMedCrossRef
20.
go back to reference Grandbarbe L, Michelucci A, Heurtaux T, Hemmer K, Morga E, Heuschling P. Notch signaling modulates the activation of microglial cells. Glia. 2007;55:1519–30.PubMedCrossRef Grandbarbe L, Michelucci A, Heurtaux T, Hemmer K, Morga E, Heuschling P. Notch signaling modulates the activation of microglial cells. Glia. 2007;55:1519–30.PubMedCrossRef
21.
go back to reference Wu YP, Tan CK, Ling EA. Expression of Fos-like immunoreactivity in the brain and spinal cord of rats following middle cerebral artery occlusion. Exp Brain Res. 1997;115:129–36.PubMedCrossRef Wu YP, Tan CK, Ling EA. Expression of Fos-like immunoreactivity in the brain and spinal cord of rats following middle cerebral artery occlusion. Exp Brain Res. 1997;115:129–36.PubMedCrossRef
22.
go back to reference Yuan Y, Zha H, Rangarajan P, Ling EA, Wu C. Anti-inflammatory effects of Edaravone and Scutellarin in activated microglia in experimentally induced ischemia injury in rats and in BV-2 microglia. BMC Neurosci. 2014;15:125.PubMedCentralPubMedCrossRef Yuan Y, Zha H, Rangarajan P, Ling EA, Wu C. Anti-inflammatory effects of Edaravone and Scutellarin in activated microglia in experimentally induced ischemia injury in rats and in BV-2 microglia. BMC Neurosci. 2014;15:125.PubMedCentralPubMedCrossRef
23.
go back to reference Thomas WE. Brain macrophages: evaluation of microglia and their functions. Brain Res Brain Res Rev. 1992;17:61–74.PubMedCrossRef Thomas WE. Brain macrophages: evaluation of microglia and their functions. Brain Res Brain Res Rev. 1992;17:61–74.PubMedCrossRef
24.
go back to reference Ling EA, Wong WC. The origin and nature of ramified and amoeboid microglia: a historical review and current concepts. Glia. 1993;7:9–18.PubMedCrossRef Ling EA, Wong WC. The origin and nature of ramified and amoeboid microglia: a historical review and current concepts. Glia. 1993;7:9–18.PubMedCrossRef
25.
go back to reference Kaur C, Ling EA, Wong WC. Transformation of amoeboid microglial cells into microglia in the corpus callosum of the postnatal rat brain: an electron microscopical study. Arch Histol Jpn. 1985;48:17–25.PubMedCrossRef Kaur C, Ling EA, Wong WC. Transformation of amoeboid microglial cells into microglia in the corpus callosum of the postnatal rat brain: an electron microscopical study. Arch Histol Jpn. 1985;48:17–25.PubMedCrossRef
26.
28.
go back to reference Sivakumar V, Ling EA, Lu J, Kaur C. Role of glutamate and its receptors and insulin-like growth factors in hypoxia induced periventricular white matter injury. Glia. 2010;58:507–23.PubMedCrossRef Sivakumar V, Ling EA, Lu J, Kaur C. Role of glutamate and its receptors and insulin-like growth factors in hypoxia induced periventricular white matter injury. Glia. 2010;58:507–23.PubMedCrossRef
29.
go back to reference Huo Y, Rangarajan P, Ling EA, Dheen ST. Dexamethasone inhibits the Nox-dependent ROS production via suppression of MKP-1-dependent MAPK pathways in activated microglia. BMC Neurosci. 2011;12:49.PubMedCentralPubMedCrossRef Huo Y, Rangarajan P, Ling EA, Dheen ST. Dexamethasone inhibits the Nox-dependent ROS production via suppression of MKP-1-dependent MAPK pathways in activated microglia. BMC Neurosci. 2011;12:49.PubMedCentralPubMedCrossRef
30.
go back to reference Dheen ST, Jun Y, Yan Z, Tay SS, Ling EA. Retinoic acid inhibits expression of TNF-alpha and iNOS in activated rat microglia. Glia. 2005;50:21–31.PubMedCrossRef Dheen ST, Jun Y, Yan Z, Tay SS, Ling EA. Retinoic acid inhibits expression of TNF-alpha and iNOS in activated rat microglia. Glia. 2005;50:21–31.PubMedCrossRef
31.
go back to reference Zhang HF, Hu XM, Wang LX, Xu SQ, Zeng FD. Protective effects of scutellarin against cerebral ischemia in rats: evidence for inhibition of the apoptosis-inducing factor pathway. Planta Med. 2009;75:121–6.PubMedCrossRef Zhang HF, Hu XM, Wang LX, Xu SQ, Zeng FD. Protective effects of scutellarin against cerebral ischemia in rats: evidence for inhibition of the apoptosis-inducing factor pathway. Planta Med. 2009;75:121–6.PubMedCrossRef
32.
go back to reference D’Acquisto F, May MJ, Ghosh S. Inhibition of nuclear factor kappa B (NF-B): an emerging theme in anti-inflammatory therapies. Mol Interv. 2002;2:22–35.PubMedCrossRef D’Acquisto F, May MJ, Ghosh S. Inhibition of nuclear factor kappa B (NF-B): an emerging theme in anti-inflammatory therapies. Mol Interv. 2002;2:22–35.PubMedCrossRef
33.
go back to reference Ang HL, Tergaonkar V. Notch and NFkappaB signaling pathways: do they collaborate in normal vertebrate brain development and function? Bioessays. 2007;29:1039–47.PubMedCrossRef Ang HL, Tergaonkar V. Notch and NFkappaB signaling pathways: do they collaborate in normal vertebrate brain development and function? Bioessays. 2007;29:1039–47.PubMedCrossRef
34.
go back to reference Schwarzer R, Dorken B, Jundt F. Notch is an essential upstream regulator of NF-kappaB and is relevant for survival of Hodgkin and Reed-Sternberg cells. Leukemia. 2012;26:806–13.PubMedCrossRef Schwarzer R, Dorken B, Jundt F. Notch is an essential upstream regulator of NF-kappaB and is relevant for survival of Hodgkin and Reed-Sternberg cells. Leukemia. 2012;26:806–13.PubMedCrossRef
35.
go back to reference Semple BD, Kossmann T, Morganti-Kossmann MC. Role of chemokines in CNS health and pathology: a focus on the CCL2/CCR2 and CXCL8/CXCR2 networks. J Cereb Blood Flow Metab. 2010;30:459–73.PubMedCentralPubMedCrossRef Semple BD, Kossmann T, Morganti-Kossmann MC. Role of chemokines in CNS health and pathology: a focus on the CCL2/CCR2 and CXCL8/CXCR2 networks. J Cereb Blood Flow Metab. 2010;30:459–73.PubMedCentralPubMedCrossRef
36.
go back to reference McManus CM, Liu JS, Hahn MT, Hua LL, Brosnan CF, Berman JW, et al. Differential induction of chemokines in human microglia by type I and II interferons. Glia. 2000;29:273–80.PubMedCrossRef McManus CM, Liu JS, Hahn MT, Hua LL, Brosnan CF, Berman JW, et al. Differential induction of chemokines in human microglia by type I and II interferons. Glia. 2000;29:273–80.PubMedCrossRef
37.
go back to reference Wang D, Wang L, Gu J, Yang H, Liu N, Lin Y, et al. Scutellarin inhibits high glucose-induced and hypoxia-mimetic agent-induced angiogenic effects in human retinal endothelial cells through reactive oxygen species/hypoxia-inducible factor-1alpha/vascular endothelial growth factor pathway. J Cardiovasc Pharmacol. 2014;64:218–27.PubMedCrossRef Wang D, Wang L, Gu J, Yang H, Liu N, Lin Y, et al. Scutellarin inhibits high glucose-induced and hypoxia-mimetic agent-induced angiogenic effects in human retinal endothelial cells through reactive oxygen species/hypoxia-inducible factor-1alpha/vascular endothelial growth factor pathway. J Cardiovasc Pharmacol. 2014;64:218–27.PubMedCrossRef
38.
go back to reference Li H, Huang D, Gao Z, Lv Y, Zhang L, Cui H, et al. Scutellarin inhibits cell migration by regulating production of alphavbeta6 integrin and E-cadherin in human tongue cancer cells. Oncol Rep. 2010;24:1153–60.PubMed Li H, Huang D, Gao Z, Lv Y, Zhang L, Cui H, et al. Scutellarin inhibits cell migration by regulating production of alphavbeta6 integrin and E-cadherin in human tongue cancer cells. Oncol Rep. 2010;24:1153–60.PubMed
39.
go back to reference Park SR, Park KS, Park YJ, Bang D, Lee ES. CD11a, CD11c, and CD18 gene polymorphisms and susceptibility to Behcet’s disease in Koreans. Tissue Antigens. 2014;84:398–404.PubMedCrossRef Park SR, Park KS, Park YJ, Bang D, Lee ES. CD11a, CD11c, and CD18 gene polymorphisms and susceptibility to Behcet’s disease in Koreans. Tissue Antigens. 2014;84:398–404.PubMedCrossRef
40.
go back to reference Aujla PK, Bora A, Monahan P, Sweedler JV, Raetzman LT. The Notch effector gene Hes1 regulates migration of hypothalamic neurons, neuropeptide content and axon targeting to the pituitary. Dev Biol. 2011;353:61–71.PubMedCentralPubMedCrossRef Aujla PK, Bora A, Monahan P, Sweedler JV, Raetzman LT. The Notch effector gene Hes1 regulates migration of hypothalamic neurons, neuropeptide content and axon targeting to the pituitary. Dev Biol. 2011;353:61–71.PubMedCentralPubMedCrossRef
Metadata
Title
Scutellarin regulates the Notch pathway and affects the migration and morphological transformation of activated microglia in experimentally induced cerebral ischemia in rats and in activated BV-2 microglia
Authors
Yun Yuan
Parakalan Rangarajan
Enci Mary Kan
Yajun Wu
Chunyun Wu
Eng-Ang Ling
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Journal of Neuroinflammation / Issue 1/2015
Electronic ISSN: 1742-2094
DOI
https://doi.org/10.1186/s12974-014-0226-z

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