Fruits extracts of Hovenia dulcis Thunb.suppresses lipopolysaccharide—stimulated inflammatory responses through nuclear factor—kappaB pathway in Raw 264.7 cells

Objective:To investigate the anti-inflammatory effects and the action mechanism of the fruits of Horenia dulcis(H.dulcis) in lipopolysaccharide(LPS)-induced mouse macrophage Raw 264.7cells.Methods:The extract of H.dulcis fruits(EHDF) were extracted with 70%ethanol.Mouse macrophages were treated with different concentrations of EHDF in the presence and absence of LPS(1 μg/mL).To demonstrate the inflammatory mediators including nitric oxide,inducible nitric oxide synthase and cyclooxygenase(COX)-2 expression levels were analyzed by usingin vitro assay systems.COX-derived pro-inflammatory cytokines including interleukin-1 β.tumor necrosis factor- α and prostaglandin F_2 were determined using ELISA kits.Cell viability,heme oxygenase-1 expression,nuclear factor-kappaB and nuclear factor F.2-related factors 2 translocation were also investigated.Results:EHDF potently inhibited the LPS-stimulated nitric oxide,inducible nitric oxide synthase.COX-2,interleukin-1 β and tumor necrosis factor- α expression in a dose-dependent manner.EHDF suppressed the phosphorylation of inhibited kappaB-alpha and p65 nuclear translocation.Treatment of macrophage cells with EHDF alone induced the heme oxygenase-1 and nuclear translocation of nuclear factor E2-reIated factor 2.Conclusions:These results suggest that the ethanol extract of H.dulcis fruit exerts its anti-inflammatory effects by inhibiting inhibited kappaBalpha phorylation and nuclear translocation of nuclear factor-kappaB.

Ethanol Extract of Lycopodium serratum Thunb. Attenuates Lipopolysaccharide-Induced C6 Glioma Cells Migration via Matrix Metalloproteinase-9 Expression

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-κB） downstream pathway. Methods： Cell viability of ELS on C6 glioma was detected by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） 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-κB, mitogen-activated protein kinase（MAPK）, inducible nictric oxide synthase（i NOS） and cyclooxygenase-2（COX-2） protein were determined by Western 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 i NOS, 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.