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.

Saeng-Kankunbi-Tang(生肝健脾汤) Protects Liver against Oxidative Damage through Activation of ERK/Nrf2 Pathway

Objective： To investigate the cytoprotective effects of Saeng-kankunbi-tang（生肝健脾汤, SKT）, a herbal prescription consisting of Artemisia capillaris and Alisma canaliculatum, and its underlying mechanism involved. Methods： In mice, blood biochemistry and histopathology were assessed in carbon tetrachloride（CCl4）-induced oxidative hepatic injury in vivo. The animal groups included vehicle-treated control, CCl4, SKT 500 mg/（kg·day） CCl4＋SKT 200 or 500 mg/（kg·day）. In Hep G2 cell, tert-butyl hydroperoxide（t BHP） induced severe oxidative stress and mitochondrial dysfunction in vitro. The cyto-protective effects of SKT were determined by 3-（4,5-dimethylthiazol）-2,5-diphenyltetrazolium bromide（MTT） assay, fluorescence activated cell sorting analysis and western blotting. Results： The administration of SKT prevented liver damage induced by CCl4 in mice, by inhibition of hepatocyte degeneration and inflammatory cell infiltration as well as plasma parameters such as alanine aminotransferase（P〈0.01）. Moreover, treatment with t BHP induced hepatocyte death and cellular reactive oxygen species production in hepatocyte cell line. However, SKT pretreatment（30–300 μg/m L） reduced this cell death and oxidative stress（P〈0.01）. More importantly, SKT inhibited the ability of t BHP to induce changes in mitochondrial membrane transition in cell stained with rhodamine 123（P〈0.01）. Furthermore, treatment with SKT induced extracellular signal-regulated kinases-mediated nuclear factor erythroid-2-related factor 2（Nrf2） activation as well as the expressions of heme oxygenase 1 and glutamate-cystein ligase catalytic, Nrf2 target genes. Conclusion： SKT has the ability to protect hepatocyte against oxidative stress and mitochondrial damage mediated by Nrf2 activation.

Polyherbal formula SC-E3 inhibits rheumatoid arthritis activity in a mouse model of type-Ⅱcollagen-induced arthritis

Objective:SC-E3 is a polyherbal formula that contains five medicinal herbs used frequently in traditional herbal medicine.In our previous study,we demonstrated the antioxidant and anti-inflammatory effects of SC-E3.The present study examined the effects of SC-E3 in a mouse model of type-II collagen-induced arthritis(CIA).Methods:In vivo,male DBA/1 J mice were immunized by intradermal injection of bovine type-II collagen and complete or incomplete Freund’s adjuvant,to induce arthritis.SC-E3 was orally administered daily for 23 days.In vitro,bone marrow-derived macrophages(BMMs)were treated with macrophage colony-stimulating factor(M-CSF)and receptor activator of nuclear factor-j B ligand(RANKL)in the absence or presence of SC-E3.Results:Administrations of SC-E3 were found to have anti-arthritic effects in the joints of CIA mice,as evidenced by reduced paw swelling,bone erosion and deformation,inflammatory cell infiltration,and inflammation in synovial membrane.SC-E3 also reduced serum levels of tumor necrosis factor-a,interleukin-1 b,aspartate aminotransferase and alanine aminotransferase.Furthermore,tartrateresistant acid phosphatase-positive osteoclast numbers in the joints were significantly lower in SC-E3-treated CIA mice than in CIA mice.In addition,the differentiations of BMMs to multinucleated osteoclasts induced by M-CSF and RANKL stimulation were dose-dependently reduced by SC-E3.Conclusion:These results suggest that SC-E3 possesses substantial anti-arthritic activity because it inhibits pro-inflammatory cytokines and osteoclastogenesis,and that SC-E3 has potential therapeutic use for the treatment of rheumatoid arthritis.