Youngheun Jee

Radioprotective effects of an acidic polysaccharide of Panax ginseng on bone marrow cells

An acidic polysaccharide of Panax ginseng (APG), so called ginsan is known to have important immunomodulatory activities. It was recently reported that APG has radioprotective effects in mice but the detailed mechanism was not fully elucidated. This study examined the effects of APG on bone marrow cells (BMs). The phenotypical and functional changes in APG-treated BMs after gamma radiation were studied. The benefit of APG on BMs damaged by gamma radiation was determined by measuring the cell viability. Using 2 different assays, a pretreatment with APG significantly increased the viability of BMs against gamma radiation. APG-treated BMs had a significantly higher amount of IL-12, which is a major cytokine for immune responses, compared with the medium-treated BMs. The expression of MHC class II molecules of APG-treated BMs was also increased, and APG-treated BMs showed significantly higher levels of allogeneic CD4+ T lymphocyte proliferation. Furthermore, APG-treated mice had a larger number of BMs after gamma radiation than the control mice, and the BMs of APG-treated mice were successfully cultured into dendritic cells, which are the representative antigen-presenting cells. Overall, this study shows that APG alters the phenotype of BMs, increases the viability and alloreactivity of BMs after gamma radiation both in vitro and in vivo. Therefore, APG may be a good candidate radioprotective agent for BMs.

Anti-inflammatory effect of fucoidan extracted from Ecklonia cava in zebrafish model.

Fucoidan extracted from Ecklonia cava had strong anti-inflammatory activities. However, the direct effects of fucoidan of E. cava on anti-inflammatory activities in vivo model remained to be determined. Therefore, the present study was designed to assess in vivo anti-inflammatory effect of fucoidan extracted from E. cava (ECF) using tail-cutting-induced and lipopolysaccharide (LPS)-stimulated zebrafish model. Treating zebrafish model with tail-cutting and LPS-treatment significantly increased the ROS and NO level. However, ECF inhibited this tail-cutting-induced and LPS-stimulated ROS and NO generation. These results show that ECF alleviated inflammation by inhibiting the ROS and NO generation induced by tail-cutting and LPS-treatment. In addition, ECF has a protective effect against the toxicity induced by LPS exposure in zebrafish embryos. This outcome could explain the potential anti-inflammatory activity of ECF, which might have a beneficial effect during the treatment of inflammatory diseases.

Molecular characteristics and anti-inflammatory activity of the fucoidan extracted from Ecklonia cava

Enzymatic extraction has been successfully used for extracting numerous biologically active compounds from a wide variety of seaweeds. In this study, we found that enzymatic extraction of the fucoidan from Ecklonia cava may be more advantageous than water extraction. Therefore, we studied the E. cava fucoidans extracted by the enzymatic extraction technique and used ion-exchange chromatography to determine their molecular characteristics and anti-inflammatory activities. The crude and fractionated fucoidans (F1, F2, and F3) consisted mostly of carbohydrates (47.1-57.1%), uronic acids (9.0-15.8%), and sulfates (16.5-39.1%), as well as varying levels of proteins (1.3-8.7%). The monosaccharide levels significantly differed, and the composition included fucose (53.1-77.9%) and galactose (10.1-32.8%), with a small amount of rhamnose (2.3-4.5%), xylose (4.0-8.2%), and glucose (0.8-2.2%). These fucoidans contained one or two subfractions with an average molecular weight (Mw) ranging from 18 to 359×10(3)g/mol. These fucoidans significantly inhibited NO production in lipopolysaccharide (LPS)-induced Raw 264.7 macrophage cells by down-regulating the expression of iNOS, COX-2, and pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1β. Thus, the present results suggest that E. cava fucoidan may be a potentially useful therapeutic approach for various inflammatory diseases.

An acidic polysaccharide of Panax ginseng ameliorates experimental autoimmune encephalomyelitis and induces regulatory T cells

An acidic polysaccharide of Panax ginseng (APG), so called ginsan, is a purified polysaccharide. APG has multiple immunomodulatory effects of stimulating natural killer (NK) and T cells and producing a variety of cytokines that proved to diminish the proinflammatory response, and protect from septic lethality. To determine APGs role in the autoimmune demyelinating disease, we tested whether APG can regulate inflammatory and encephalitogenic response in experimental autoimmune encephalomyelitis (EAE), an animal model of human multiple sclerosis (MS). Here, we demonstrate the therapeutic efficacy of the APG which induces the suppression of an encephalitogenic response during EAE. APG significantly ameliorates the progression of EAE by inhibiting the proliferation of autoreactive T cells and the production of inflammatory cytokines such as IFN-γ, IL-1β and IL-17. More importantly, APG promotes the generation of immunosuppressive regulatory T cells (Tregs) through the activation of transcription factor, Foxp3. Furthermore, the depletion of CD25+ cells from APG-treated EAE mice abrogates the beneficial effects of EAE. The capacity of APG to induce clinically beneficial effects furthers our understanding of the basis for its therapeutic immunosuppression of EAE and, possibly, MS. Thus, our results suggest that APG may serve as an effective therapy for MS and other autoimmune diseases.

Acidic polysaccharide of Panax ginseng as a defense against small intestinal damage by whole-body gamma irradiation of mice.

An acidic polysaccharide of Panax ginseng (APG), ginsan, has been reported to protect the hematopoietic system by increasing the number of bone marrow cells and spleen cells. Therefore, we evaluated the ability of APG to protect mice from radiation-induced damage of the small intestine. APG treatment caused the lengthening of villi and a numerical increase of crypt cells in the small intestine at 3.5 days after 7Gy irradiation compared to irradiated, non-treated controls. In addition, APG significantly inhibited irradiation-induced apoptosis by decreasing the amount of pro-apoptotic p53 and Bax as well as augmenting that of anti-apoptotic Bcl-2 at 24h after irradiation. These results indicate that APG might be a useful adjunct to therapeutic irradiation as a protective agent for the gastrointestinal tract of cancer patients.

Lipocalin-2 Protein Deficiency Ameliorates Experimental Autoimmune Encephalomyelitis THE PATHOGENIC ROLE OF LIPOCALIN-2 IN THE CENTRAL NERVOUS SYSTEM AND PERIPHERAL LYMPHOID TISSUES

Background: The role of LCN2 in EAE is not clear. Results: LCN2 expression increased in EAE. Lcn2 deficiency attenuated EAE symptoms and pathology. LCN2 enhanced glial expression of inflammatory mediators and peripheral encephalitogenic T cell activation in vitro and in vivo. Conclusion: Both central and peripheral LCN2 contributed to EAE development. Significance: LCN2 can be targeted for treatment of multiple sclerosis. Lipocalin-2 (LCN2) plays an important role in cellular processes as diverse as cell growth, migration/invasion, differentiation, and death/survival. Furthermore, recent studies indicate that LCN2 expression and secretion by glial cells are induced by inflammatory stimuli in the central nervous system. The present study was undertaken to examine the regulation of LCN2 expression in experimental autoimmune encephalomyelitis (EAE) and to determine the role of LCN2 in the disease process. LCN2 expression was found to be strongly increased in spinal cord and secondary lymphoid tissues after EAE induction. In spinal cords astrocytes and microglia were the major cell types expressing LCN2 and its receptor 24p3R, respectively, whereas in spleens, LCN2 and 24p3R were highly expressed in neutrophils and dendritic cells, respectively. Furthermore, disease severity, inflammatory infiltration, demyelination, glial activation, the expression of inflammatory mediators, and the proliferation of MOG-specific T cells were significantly attenuated in Lcn2-deficient mice as compared with wild-type animals. Myelin oligodendrocyte glycoprotein-specific T cells in culture exhibited an increased expression of Il17a, Ifng, Rorc, and Tbet after treatment with recombinant LCN2 protein. Moreover, LCN2-treated glial cells expressed higher levels of proinflammatory cytokines, chemokines, and MMP-9. Adoptive transfer and recombinant LCN2 protein injection experiments suggested that LCN2 expression in spinal cord and peripheral immune organs contributes to EAE development. Taken together, these results imply LCN2 is a critical mediator of autoimmune inflammation and disease development in EAE and suggest that LCN2 be regarded a potential therapeutic target in multiple sclerosis.

Peroxisome proliferator–activated receptor γ–mediated suppression of dendritic cell function prevents the onset of atopic dermatitis in NC/Tnd mice

BACKGROUNDnDendritic cells (DCs) are one of the key regulators for the initiation of allergic responses in patients with atopic dermatitis (AD), being strongly triggered by epithelial cell-derived thymic stromal lymphopoietin (TSLP). Because peroxisome proliferator-activated receptor (PPAR) γ acts as a negative regulator in immune cells, suppressive properties of PPARγ in allergic responses have been proposed.nnnOBJECTIVEnBecause pieces of evidence must be organized to identify the exact role of PPARγ in immune regulation, we explored the suppressive effects of a PPARγ agonist on various functions of DCs and the onset of AD in a murine model.nnnMETHODSnEffects of rosiglitazone (RSG) on DCs that were derived from NC/Tnd mice, a model for human AD, were analyzed. RSG was administered to NC/Tnd mice to evaluate its preventive and therapeutic effects on the development of AD.nnnRESULTSnRSG inhibited TSLP-induced DC maturation through downregulation of costimulatory molecules. TSLP-promoted expressions of chemokines in DCs were also suppressed by RSG treatment. Moreover, we showed the necessity of matrix metalloproteinase 9 in TSLP-promoted DC migration by using DCs derived from matrix metalloproteinase 9-deficient NC/Tnd mice, as well as the suppressive effect of PPARγ in the process. Daily oral administration of RSG to NC/Tnd mice before the onset of AD revealed a significant reduction in severity of skin lesions and scratching behavior. In mice treated with RSG, both expression of TSLP in the skin and maturation and migration of DCs were markedly suppressed.nnnCONCLUSIONnPPARγ can be provided as an inhibitory regulator of TSLP-stimulated DCs in the onset of allergic reactions.

Enzymatic Extract from Ecklonia cava Induces the Activation of Lymphocytes by IL-2 Production Through the Classical NF-κB Pathway

Activated nuclear factor-kappa B (NF-κB), a well-known transcription factor, leads to the development, differentiation, and proliferation of T and B lymphocytes and the secretion of cytokines by the classical pathway. We have examined here whether an enzymatic extract (ECK) from the brown seaweed, Alariaceae Laminariales Ecklonia cava may contribute to activating lymphocytes through the NF-κB pathway for participation in immune responses. In our study, ECK dose-dependently enhanced the proliferation of lymphocytes. ECK significantly increased the phosphorylation of inhibitors of κB at 0.25 and 0.5xa0h of exposure, followed by its gradual decrease. In addition, NF-κB p65 was gradually activated, and its binding to nuclear deoxyribonucleic acid was observed from 0.25xa0h after stimulation (up to 0.5xa0h). Further experiments showed that the application of N-p-tosyl-l-phenylalanine chloromethyl ketone, an NF-κB inhibitor, significantly blocked ECK-induced lymphocytes proliferation and the interleukin (IL)-2 productions. Accordingly, our results suggest that ECK increases the production of IL-2 through the activation of NF-κB then induces the proliferation of lymphocytes with the coordinated stimulation of IL-2.

Bee Venom Acupuncture Alleviates Experimental Autoimmune Encephalomyelitis by Upregulating Regulatory T Cells and Suppressing Th1 and Th17 Responses.

The protective and therapeutic mechanism of bee venom acupuncture (BVA) in neurodegenerative disorders is not clear. We investigated whether treatment with BVA (0.25 and 0.8xa0mg/kg) at the Zusanli (ST36) acupoints, located lateral from the anterior border of the tibia, has a beneficial effect in a myelin basic protein (MBP)68–82-induced acute experimental autoimmune encephalomyelitis (EAE) rat model. Pretreatment (every 3xa0days from 1xa0h before immunization) with BVA was more effective than posttreatment (daily after immunization) with BVA with respect to clinical signs (neurological impairment and loss of body weight) of acute EAE rats. Treatment with BVA at the ST36 acupoint in normal rats did not induce the clinical signs. Pretreatment with BVA suppressed demyelination, glial activation, expression of cytokines [interferon (IFN)-γ, IL-17, IL-17A, tumor necrosis factor-alpha (TNF-α), and IL-1β], chemokines [RANTES, monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein (MIP)-1α], and inducible nitric oxide synthase (iNOS), and activation of p38 mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB (p65 and phospho-IκBα) signaling pathways in the spinal cord of acute EAE rats. Pretreatment with BVA decreased the number of CD4+, CD4+/IFN-γ+, and CD4+/IL-17+ T cells, but increased the number of CD4+/Foxp3+ T cells in the spinal cord and lymph nodes of acute EAE rats. Treatment with BVA at six placebo acupoints (SP9, GB39, and four non-acupoints) did not have a positive effect in acute EAE rats. Interestingly, onset and posttreatment with BVA at the ST36 acupoint markedly attenuated neurological impairment in myelin oligodendrocyte glycoprotein (MOG)35–55-induced chronic EAE mice compared to treatment with BVA at six placebo acupoints. Our findings strongly suggest that treatment with BVA with ST36 acupoint could delay or attenuate the development and progression of EAE by upregulating regulatory T cells and suppressing T-helper (Th) 17 and Th1 responses. These results warrant further investigation of BVA as a treatment for autoimmune disorders of the central nervous system.

Geraniin down regulates gamma radiation-induced apoptosis by suppressing DNA damage.

Gamma ray irradiation triggers DNA damage and apoptosis of proliferating stem cells and peripheral immune cells, resulting in the destruction of intestinal crypts and lymphoid system. Geraniin is a natural compound extracts from an aquatic plant Nymphaea tetragona and possesses good antioxidant property. In this study, we demonstrate that geraniin rescues radiosensitive splenocytes and jejunal crypt cells from radiation-induced DNA damage and apoptosis. Isolated splenocytes from C57BL/6 mice treated with geraniin were protected against radiation injury of 2 Gy irradiation through the enhancement of the proliferation and attenuation of DNA damage. Also, geraniin inhibited apoptosis in radiosensitive splenocytes by reducing the expression level and immunoreactivity of proapoptotic p53 and Bax and increasing those of anti-apoptotic Bcl-2. In mice exposed to radiation, geraniin treatment protected splenocytes and intestinal crypt cells from radiation-induced cell death. Our results suggest that geraniin presents radioprotective effects by regulating DNA damage on splenocytes, exerting immunostimulatory capacities and inhibiting apoptosis of radiosensitive immune cells and jejunal crypt cells. Therefore, geraniin can be a radioprotective agent against γ-irradiation exposure.