LPS-induced iNOS expression in N9 microglial cells is suppressed by geniposide via ERK, p38 and nuclear factor-κB signaling pathways

Zhang,Gu; He,Jun-Lin; Xie,Xiao-Yan; Yu,Chao

doi:10.3892/ijmm.2012.1030

LPS-induced iNOS expression in N9 microglial cells is suppressed by geniposide via ERK, p38 and nuclear factor-κB signaling pathways

Authors:
- Gu Zhang
- Jun-Lin He
- Xiao-Yan Xie
- Chao Yu
View Affiliations

Affiliations: Department of Pathology, Zhejiang Cancer Hospital, Zhejiang 310022, P.R. China, Laboratory of Reproductive Biology, School of Public Health, Chongqing Medical University, Chongqing 400016, P.R. China, Institute of Life Sciences, Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: June 14, 2012 https://doi.org/10.3892/ijmm.2012.1030
Pages: 561-568

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Activated microglia producing reactive nitrogen species, inflammatory factors, reactive oxygen species (ROS) and other neurovirulent factors, can lead to the development of neurodegenerative diseases. Certain compounds can inhibit the activation of microglia. However, the mechanisms remain unclear. In the present study, we investigated the inhibitory effect of geniposide on the production of ROS and inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-stimulated N9 murine microglial cells through the p38, ERK1/2 and nuclear factor-κB (NF-κB) signaling pathways. After the N9 cells were pre-treated with the vehicle or geniposide and exposed to LPS for the time indicated, the MTT conversion test was used to assess cell viability. Suitable concentrations were chosen and adjusted according to the experiments. Extracellular nitric oxide (NO) release was measured by Griess reaction. The formation of ROS and intracellular NO was evaluated by fluorescence imaging. NOS activities were determined using commercially available kits. The morphology of the N9 cells was examined by hematoxylin and eosin staining. The expression of iNOS mRNA was examined by RT-PCR. The protein levels of iNOS, p38 mitogen-activated protein kinase (MAPK), ERK1/2 and NF-κB, inhibitory factor-κB-α (IκB-α) were determined by western blot analysis. The results showed that geniposide attenuated the activation of N9 cells and inhibited the overproduction of NO, intracellular ROS and the expression of iNOS induced by LPS in the cells. In addition, geniposide blocked the phosphorylation of p38, ERK1/2 and inhibited the drop-off of IκB induced by LPS in the cells. These data indicate that geniposide has therapeutic potential for the treatment of neurodegenerative diseases, and that it exerts its effects by inhibiting inflammation.

View Figures

View References

1.	WF HickeyH KimuraPerivascular microglial cells of the CNS are bone marrow derived and present antigen in vivoScience239290292198810.1126/science.32760043276004
2.	RB RockG GekkerS HuRole of microglia in central nervous system infectionsClin Microbiol Rev17942964200410.1128/CMR.17.4.942-964.200415489356
3.	H WilmsL ZeccaP RosenstielInflammation in Parkinson’s diseases and other neurodegenerative diseases: cause and therapeutic implicationsCurr Pharm Des13192519282007
4.	J PalaceInflammation versus neurodegeneration: consequences for treatmentJ Neurol Sci2594649200710.1016/j.jns.2006.05.07217418237
5.	YS KimTH JohMicroglia, major player in the brain inflammation: their roles in the pathogenesis of Parkinson’s diseaseExp Mol Med38333347200616953112
6.	J RogersD MastroeniB LeonardNeuroinflammation in Alzheimer’s disease and Parkinson’s disease: are microglia pathogenic in either disorder?Int Rev Neurobiol822352462007
7.	PL McGeerEG McGeerGlial reactions in Parkinson’s diseaseMov Disord234744832008
8.	L QinY LiuT WangNADPH oxidase mediates lipopolysaccharide-induced neurotoxicity and proinflammatory gene expression in activated microgliaJ Biol Chem27914151421200410.1074/jbc.M30765720014578353
9.	XL BiJY YangYX DongResveratrol inhibits nitric oxide and TNF-alpha production by lipopolysaccharide-activated microgliaInt Immunopharmacol585193200515589480
10.	YM ZhuNS AzahriD YuPJ WollEffects of COX-2 inhibition on expression of vascular endothelial growth factor and interleukin-8 in lung cancer cellsBMC Cancer8218200810.1186/1471-2407-8-21818671849
11.	RN SahaK PahanRegulation of inducible nitric oxide synthase gene in glial cellsAntioxid Redox Signal8929947200610.1089/ars.2006.8.92916771683
12.	T AkaoK KobayashiM AburadaEnzymic studies on the animal and intestinal bacterial metabolism of geniposideBiol Pharm Bull1715731576199410.1248/bpb.17.15737735197
13.	SW WangCY LaiCJ WangInhibitory effect of geniposide on aflatoxin B1-induced DNA repair synthesis in primary cultured rat hepatocytesCancer Lett65133137199210.1016/0304-3835(92)90157-Q1511417
14.	CH PengCN HuangSP HsuCJ WangPenta-acetyl geniposide-induced apoptosis involving transcription of NGF/p75 via MAPK-mediated AP-1 activation in C6 glioma cellsToxicology238130139200710.1016/j.tox.2007.05.02917651887
15.	JH LiuF YinLX GuoXH DengYH HuNeuroprotection of geniposide against hydrogen peroxide induced PC12 cells injury: involvement of PI3 kinase signal pathwayActa Pharmacol Sin30159165200910.1038/aps.2008.2519151742
16.	M YamazakiK ChibaNeurotrophic effects of genipin on Neuro2a cellsJ Health Sci51687692200510.1248/jhs.51.687
17.	KN NamYS ChoiHJ JungGenipin inhibits the inflammatory response of rat brain microglial cellsInt Immunopharmacol10493499201010.1016/j.intimp.2010.01.01120123040
18.	T MosmannRapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assaysJ Immunol Methods655563198310.1016/0022-1759(83)90303-46606682
19.	M LantornoH ChenJA KimGhrelin has novel vascular actions that mimic PI 3-kinase-dependent actions of insulin to stimulate production of NO from endothelial cellsAm J Physiol Endocrinol Metab292756764200710.1152/ajpendo.00570.200617106060
20.	L MarcocciJJ MaguireMT Droy-LefaixL PackerThe nitric oxide-scavenging properties of Ginkgo biloba extract EGb 761Biochem Biophys Res Commun201748755199410.1006/bbrc.1994.17648003011
21.	F Gonzalez-ScaranoG BaltuchMicroglia as mediators of inflammatory and degenerative diseasesAnnu Rev Neurosci22219240199910.1146/annurev.neuro.22.1.21910202538
22.	G StollS JanderThe role of microglia and macrophages in the pathophysiology of the CNSProg Neurobiol58233247199910.1016/S0301-0082(98)00083-510341362
23.	GW KreutzbergMicroglia: a sensor for pathological events in the CNSTrends Neurosci19312318199610.1016/0166-2236(96)10049-78843599
24.	L MedaMA CassatellaGI SzendreiActivation of microglial cells by β-amyloid protein and IFN-γNature3746476501995
25.	B MayerK SchmidtP HumbertE BöhmeBiosynthesis of endothelium-derived relaxing factor: a cytosolic enzyme in porcine aortic endothelial cells Ca²⁺-dependently converts L-arginine into an activator of soluble guanylyl cyclaseBiochem Biophys Res Commun164678685198910.1016/0006-291X(89)91513-12573351
26.	A Bal-PriceGC BrownInflammatory neurodegeneration mediated by nitric oxide from activated glia-inhibiting neuronal respiration, causing glutamate release and excitotoxicityJ Neurosci21648064912001
27.	G ZalbaA FortunoG San JoseOxidative stress, endothelial dysfunction and cerebrovascular diseaseCerebrovasc Dis24Suppl 1S24S29200710.1159/000107376
28.	HW JungCH YoonKM ParkHexane fraction of Zingiberis Rhizoma Crudus extract inhibits the production of nitric oxide and proinflammatory cytokines in LPS-stimulated BV2 microglial cells via the NF-kappaB pathwayFood Chem Toxicol47119011972009
29.	HL PahlActivators and target genes of Rel/NF-κB transcription factorsOncogene18685368661999