Ethanol Extract of Lycopodium serratum Thunb.Attenuates Lipopolysaccharide-Induced C6 Glioma Cells M

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Objectives:To elucidate how ethanol extract of L.serratum (ELS) could exert anti-migratory effects on glioma with the suppression of nuclear factor kappa B (NF-κ B) downstream pathway.Methods:Cell viability of ELS on C6 glioma was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyttetrazolium bromide (MTTr) assay.Nitric oxide (NO) assay and 2\',7\'-dichlorofluorescin diacetate (DCFH-DA) assay were applied to measure NO production and reactive oxygen species (ROS) generation on lipopolysaccharide (LPS)-induced C6 glioma cells.NF-κ B,mitogen-activated protein kinase (MAPK),inducible nictric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2)protein were determined by Westem blot.Wound healing assay was used to investigate the inhibitory effect of ELS on fetal bovine serum (FBS)-induced migration and matrix metalloproteinase (MMP)-9 and-2 activity was examined by zymography.Results:ELS suppressed LPS-induced phosphorylation of extracellular signal-regulated kinase (ERK),c-Jun N-terminal kinase (JNK),and p38 through inhibiting the expression of chemokine CCL2 (or monocyte chemoattractant protein-1,MCP-1).In addition,ELS inhibited the expression of iNOS,COX-2,and the production of NO by LPS in C6 glioma cells.ELS also significantly decreased serum-induced migration of C6 glioma cells in scratch wound healing in a dose-dependent manner (P<0.01).The activity of MMP-9 and-2 were also significantly attenuated by ELS with LPS treatment (P<0.01).Conclusion:Our results suggest that downregulation of MMP-9 gene expression might be involved in the anti-migration effect of ELS against LPS-induced C6 glioma cells.
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