• 동의생리병리학회지
• /
• 제27권4호
• /
• pp.431-436
• /
• 2013

Bone homeostasis is maintained by co-ordination of bone-resorbing osteoclasts and bone-forming osteoblasts. Imbalance between osteoclasts and osteoblasts leads to many bone diseases such as osteoporosis, rheumatoid arthritis. Taxillus chinensis is a herb that has been widely used to improve bone health. However, the effect and mechanism of Taxillus chinensis extract on osteoclast differentiation and bone resportion has been unknown. Thus, We investigated the effect of Taxillus chinensis on expression of receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation and bone resorption. Also, the action of Taxillus chinensis on mechanisms relating to osteoclast differentiation was studied. In this results, we identified that Taxillus chinensis significantly inhibited RANKL-induced osteoclast differentiation and bone resportion. Moreover, Taxillus chinensis was suppressed the activation of NF-${\kappa}B$ in bone marrow macrophage treated RANKL and M-CSF. Taxillus chinensis was down-regulated the mRNA expression of c-Fos, nuclear factor of activated T-cells (NFAT)c1, osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphatase (TRAP). The cell adhesion-related molecules such as integrin ${\alpha}v$ and integrin ${\beta}3$, and the filamentous actin (F-actin) rings of mature osteoclasts-related molecules such as dendritic cell-specific transmembrane preotein (DC-STAMP) and cathepsin K are also suppressed. Taken together, these results indicated that Taxillus chinensis will be a good candidate to treat osteoclast-mediated bone diseases.

• 대한약침학회지
• /
• 제6권2호
• /
• pp.45-56
• /
• 2003

Objective : The purpose of this study was to investigate the effects of Bee Venom on the lipopolysaccharide(LPS), sodium nitroprusside(SNP), hydrogen peroxide$(H_2O_2)$-induced expression inducilble nitric oxide synthetase(iNOS), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) and nuclear factor kappa B(NF-kB) in RAW 264.7 cells, a murine macrophage cell line. Method : The expressions of expression iNOS and TNF-${\alpha}$ were determined by western blotting with corresponding antibodies. The expressions of expression NF-kB was assayed by EMSA method. Results : 1. The 0.5, 1 and $5{\mu}g/mg$ of bee venom on LPS-induced expression of iNOS, the $5{\mu}g/mg$ of bee venom on SNP-induced expression of iNOS and the $1{\mu}g/mg$ of bee venom on $H_2O_2$-induced expression of iNOS compared with control were inhibited significantly. 2. The 0.5, 1 and $5{\mu}g/mg$ of bee venom inhibited significantly LPS and $H_2O_2$-induced expression of TNF-${\alpha}$ compared with control, respectively. The $0.5{\mu}g/mg$ of bee venom increased significantly SNP-induced expression of TNF-${\alpha}$ compared with control. 3. The $5{\mu}g/mg$ of bee venom on LPS-induced expression of NF-kB, the $0.5{\mu}g/mg$ of bee venom on SNP-induced expression of NF-kB and the 0.5, $5{\mu}g/mg$ of bee venom on $H_2O_2$-induced expression of NF-kB were inhibited significantly compared with control, respectively.

• Journal of Acupuncture Research
• /
• 제21권5호
• /
• pp.179-190
• /
• 2004

Objectives : The purpose of this study was to investigate the effect of Bee Venom and Melittin acupuncture solution on the lipopolysaccharide and sodium nitroprusside- induced expression of cytosolic phospholipase $A_2$, tumor necrosis factor-${\alpha}$ and calcium concentration in RAW 264.7 cells, a murine macrophage cell line. Methods : The expression of cytosolic phospholipase $A_2$ and tumor necrosis factor-${\alpha}$ was determined by western blotting with corresponding antibodies, and the generation of intracellular calcium concentration was investigated by delta scan system in RAW 264.7 cells. Results : 1. Compared with control, expressions of lipopolysaccharide-induced cytosolic phospholipase $A_2$ were decreased significantly by $5{\mu}g/mL$ of bee venom and 5, $10{\mu}g/mL$ of melittin acupuncture solution and decreased by 0.5, $1{\mu}g/mL$ of bee venom. 2. Compared with control, expressions of sodium nitroprusside-induced phospholipase $A_2$ were decreased significantly by 0.5, 1, $5{\mu}g/mL$ of bee venom and by 5, $10{\mu}g/mL$ of melittin acupuncture solution. 3. Compared with control, expressions of lipopolysaccharide-induced tumor necrosis factor-${\alpha}$ were decreased significantly by $10{\mu}g/mL$ of melittin acupuncture solution and were not changed significantly by 0.5, 1, $5{\mu}g/mL$ of bee venom and $5{\mu}g/mL$ of melittin acupuncture solution. 4. Compared with control, expressions of sodium nitroprusside-induced tumor necrosis factor-${\alpha}$ were decreased significantly by 1, $5{\mu}g/mL$ of bee venom and 5, $10{\mu}g/mL$ of melittin acupuncture solution and decreased by $0.5{\mu}g/mL$ of bee venom 5. Compared with control, lipopolysaccharide-induced intracellular calcium concentrations were decreased by 0.5, 1, $5{\mu}g/mL$ of bee venom and $10{\mu}g/mL$ of melittin acupuncture solution and increased by $5{\mu}g/mL$ of melittin acupuncture solution. 6. Compared with control, sodium nitroprusside-induced intracellular calcium concentrations were decreased by 0.5, 1, $5{\mu}g/mL$ of bee venom and 5, $10{\mu}g/mL$ of melittin acupuncture solution.

• 동의생리병리학회지
• /
• 제23권5호
• /
• pp.1106-1115
• /
• 2009

Mori Ramulus has multiple applications in Korean traditional medicine prescription because it has antioxidant and anti-inflammatory effects by reducing macrophage activities. Yet, no studies on the anti-arthritic activity of EMR (extract of Mori Ramulus) have been reported in vitro and in vivo. Rheumatoid arthritis (RA) is a systemic autoimmune disease with chronic inflammation characterized by hyperplasia of synovial cells in affected joints, which ultimately leads to the destruction of cartilage and bone. Because collagen-induced arthritis (CIA) is similar to RA in pathological symptoms and immune reactions, there have been several reports concerning RA using CIA mouse model. Here, we investigated the effects of Mori Ramulus on RA using CIA mice. The importance of CD4+ Th1 cells in RA progress was previously indicated and studies further showed that Th17 cells play a prime role in severity of disease. Accordingly, the present study was focused on CIA associated with CD4+ Th1 cells and Th1 7 cells. DBA/1OlaHsd mice were immunized with bovine type II collagen (CII). After a second collagen immunization, mice were treated with EMR once a day for 4 weeks. The severity of arthritis within the paw joints was evaluated by histological assessment of cartilage destruction and pannus formation. Immune cells in peripheral blood mononuclear cells (PBMC), draining lymph node (DLN) and paw joints, cytokine production and gene expression were assessed from CIA mouse using ELISA, FACS and real-time PCR analysis. Administration of EMR significantly suppressed the progression of CIA and inhibited the production of TNF-$\alpha$, IL-6 and IL-17 in the serum. The erosion of cartilage was dramatically reduced in mouse knees after treatment with EMR. In conclusion, our results demonstrate that EMR significantly suppressed the progression of CIA and that this action was mediated by the decreased production of TNF-$\alpha$, IL-6, IL-17 and collagen II-specific antibody in the serum. EMR suppressed Th17 cells and reduced level of IL-6 via B cell suppression, and thus, the levels of autoantibodies produced from B cells were decreased. Furthermore, EMR suppressed NKT cells which directly stimulate B cells and develop imbalance of Th1/Th2 cell. Oral administration of EMR (100 mg/kg or 200 mg/kg) significantly suppressed the progression of CIA, which is comparable to that of methotrexate (MTX, 0.3 mg/kg) used as a positive control. We are currently studying the mechanism underlying the therapeutic role for EMR in CIA mice.

• 한국식품과학회지
• /
• 제49권3호
• /
• pp.343-348
• /
• 2017

상백피 및 상지 에탄올 추출물의 항염증 효과를 확인하기 위하여 RAW 264.7 세포를 사용하여 시험하였다. 상백피, 상지 추출물의 활성산소종, 산화질소(II), 프로스타글란딘 $E_2$, 인터류킨-6, 종양괴사인자-${\alpha}$ 등의 생성능과 COX-2, iNOS mRNA 발현을 확인하였다. 산화질소(II) 생성량은 두 추출물 모두 유의하게 비슷한 정도로 감소시켰으나, 활성산소종 생성에서는 상백피 추출물이 상지 추출물 보다 억제 효과가 높게 나타났다. 프로스타글란딘 $E_2$와 인터류킨-6 생성에서는 상지 추출물의 억제 효과가 강하게 나타났으며, COX-2와 iNOS mRNA 발현량 또한 상지 추출물의 상백피에 비해 유의하게 감소시킴을 확인하였다. 본 연구에서 확인한 결과로 상백피, 상지 추출물 모두 염증 매개인자 발현 억제를 통해 항염증 효과를 가짐을 알 수 있었으며, 상지 추출물이 상백피 추출물 보다 염증 완화 효과가 강한 것으로 나타났다. 향후에는 상백피 및 상지 추출물의 주요 활성 성분들의 차이와, 염증완화 메커니즘을 통한 관련 질환에 효과가 있는지 정확한 작용메커니즘 확인 등의 분석실험이 필요할 것으로 생각된다.

• Natural Product Sciences
• /
• 제13권1호
• /
• pp.33-39
• /
• 2007

Eleven polyhydroxyursane triterpenoids (PHUTs) were tested to determine their cytoprotective, immunosuppressive and anti-inflammatory effects. To compare the bioactivities of $19{\alpha}$-hydroxyursane-type triterpenoids {23-hydroxytormentic acid (6), its methyl ester (7), tormentic acid (8), niga-ichigoside $F_1$ (9),euscaphic acid (10) and kaji-ichigoside $F_1$ (11)} of the Rosaceae crude drugs (Rubi Fructus and Rosa rugosae Radix) with PHUTs possessing no $19{\alpha}-hydroxyl$ of Centella asiatica (Umbelliferae), the four PHUTs, asiaticoside (1), madecassoside (2), asiatic acid (3), and madecassic acid (4) were isolated from C. asiatica and 23-hydroxyursolic acid (5) from Cussonia bancoensis. Cytoprotective effects were assessed by measuring cell viabilities against cisplatin-induced cytotoxocity in $LLC-PK_1$, cells (proximal tubule, pig kidney) to determine whether these agents have protective effects against nephrotoxicity caused by cisplatin. The inhibitory effect of 11 PHUTS on nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ were evaluated by measuring nitrite accumulation in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells, and their anti-inflammatory effects were tested in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model. Six MHUTs (compounds 1, 2, 4, 6, 10, and 11) exhibited higher cell viabilities during cisplatin-induced cytotoxicity testing even at a concentration of $200\;{\mu}g/ml$ than cisplatin only-treated group, suggesting that ese compounds have the potentcytoprotective efffcts. Compounds 1 and 3 of the C. asiatica and niga-ichigoside $F_1$ exhibited no inhibitory effect on NO and/or $PGE_2$ production whereas other PHUTs produced mild to significant NO and/or $PGE_2$ production.The four compounds (2, 5, 9, and 10) potently inhibited mouse ear edema induced by TPA whereas two compounds (1 and 3) had no activity in this test. These results suggest that many PHUTs are potentchemopreventives. Structure-activity relationship (SAR) was also discussed in each assay with regard to the significant role of OHs at the position of 2, 3, 6, 19, and 23 and to the glycoside linkage at the 28-carboxyl.

• 한국동물위생학회지
• /
• 제42권4호
• /
• pp.201-216
• /
• 2019

Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failure in sows and respiratory distress in all age pigs. Porcine respiratory disease complex (PRDC) is a disease caused by opportunistic bacterial infection secondary to a weakened immune system by a preceding respiratory infection. In this study, we tried to compare the immune responses in PRRS and PRDC groups to clearly characterize the disease severity. Eighty-five pigs were infected with various Korean field PRRS virus strains. Infected animals were classified into PRRS (n=32) and PRDC (n=53) groups based on lung lesions such as interstitial pneumonia, suppurative pneumonia, and pleuropneumonia. The immune cell population of bronchoalveolar lavage cells (BALc) was evaluated on 14 and 28 days post infection (dpi) and PMBC cytokine expression was measured on 0, 3, 7, 14 dpi to investigate early inflammatory reactions. Pulmonary lesion severity was negatively correlated with alveolar macrophage (AM) in both PRRS and PRDC groups on 14 and 28 dpi. AM in BALc was less populated in PRDC group on 28 dpi compared to PRRS group. AM in BALc was significantly less populated in PRDC group on 28 dpi compared to 14 dpi. In addition, cytotoxic T lymphocyte (CTL) in BALc was higher populated in PRDC group on 14 dpi and 28 dpi compared to PRRS group. In the case of PBMC cytokine TNF-α, IFN-α, IL-1β, IFN-γ, FoxP3, and IL-2, the PRRS group showed higher expression than the PRDC group on 7 dpi, 14 dpi, 7 dpi, 14 dpi, 14 dpi, and 14 dpi, respectively. On the other hand, in the case of IFN-β, IL-6, IL-8, IL-4, and IL-17, the PRDC group showed higher PBMC cytokine expression at 14 dpi, 7 dpi, 14 dpi, 3 dpi, and 3 dpi, respectively, than the PRRS group. Based on these results, our study could characterize differential immune responses in pigs with PRRS or PRDC.

• 한국식품영양학회지
• /
• 제30권1호
• /
• pp.96-104
• /
• 2017

To obtain the immunomodulating polysaccharide from chaga mushroom (Inonotus obliquus sclerotia, IO), crude polysac- charide fractions (IO-M-CP and IO-CP, respectively) prepared from hot-water extract (IO-W) of I. obliquus by EtOH precipitation after MeOH reflux or not. After IO-W was re-dissolved in water followed by EtOH addition in the case without MeOH reflux, EtOH mixture was fractionated into EtOH-soluble (IO-E) and crude polysaccharide (IO-CP). In the meanwhile, MeOH-soluble fraction (IO-M) was separated from IO-W after MeOH reflux. The residue was dissolved in water and was added by EtOH, and then EtOH mixture was also fractionation into EtOH-soluble (IO-M-E) and crude polysaccharide (IO-M-CP). As a result of the macrophage stimulating activity of these fractions, IO-CP and IO-M-CP showed significantly increased cell proliferation and cytokines production than IO-W. Particularly, IO-M-CP promotes the production of IL-12 more than IO-CP. In the splenocytes proliferating activity and intestinal immune system modulating activity through Peyer's patch, both of 2 crude polysaccharide fractions were significantly promoted in cell proliferation and cytokines production than IO-W, and IO-M-CP was more potent than IO-CP in IL-2 production from splenocytes and GM-CSF production ($10{\mu}g/mL$) in Peyer's patch cells. In addition, immunomodulating polysaccharide fractions (IO-M-CP and IO-CP) prepared from IO-W by EtOH precipitation with or without EtOH reflux showed no significant difference in the chemical composition and component sugar. These results suggested that MeOH reflux might exclude low-molecular weight materials from IO-W and consequently increase the immunomodulating activity of IO-M-CP. Therefore, it was confirmed that immunomodulation of polysaccharide prepared from hot-water extract of chaga mushroom was enhanced by fractionation including MeOH reflux and EtOH precipitation.

• Nutrition Research and Practice
• /
• 제11권6호
• /
• pp.445-451
• /
• 2017

BACKGROUND/OBJECTIVES: Endoplasmic reticulum (ER) stress is positively associated with atherosclerosis via elevating macrophage cell death and plaque formation, in which oxidative stress plays a pivotal role. Antioxidative, lipid-lowering, and anti-atherogenic effects of kimchi, a Korean fermented vegetable, have been established, wherein capsaicin, ascorbic acid, quercetin, 3-(4'-hydroxyl-3',5'-dimethoxyphenyl)propionic acid, and lactic acids were identified. In this study, mechanisms of action of kimchi methanol extracts (KME) on fatty streak formation via suppression of ER stress and apoptosis in aorta were examined in low-density lipoprotein receptor knockout mice. MATERIALS AND METHODS: Mice fed a high cholesterol diet with an oral administration of KME (KME group, $200 mg{\cdot}kg-bw^{-1}{\cdot}day^{-1}$) or distilled water (control group) for 8 weeks (n = 20 for group). Plasma lipid and oxidative stress levels were evaluated. Protein expression was measured by western blot assay. Fatty streak lesion size and the degree of apoptosis were examined in the aorta. RESULTS: Compared to the control group, in the KME group, plasma lipids levels were decreased and oxidative stress was alleviated (P < 0.05). Protein expression levels of nuclear factor (erythroid-derived 2)-like 2-mediated antioxidants in aorta were increased whereas those for ER stress markers, glucose regulated protein 78, phospho-protein kinase RNA-like ER kinase, phospho-eukaryotic initiation factor 2 subunit ${\alpha}$, X-box binding protein 1, and C/EBP homologous protein were decreased in the KME group (P < 0.05). Moreover, apoptosis was suppressed via downregulation of phospho-c-Jun N-terminal kinase, bcl-2-associated X protein, caspases-9, and -3 with a concomitant upregulation of anti-apoptotic protein, B-cell lymphoma 2 (P < 0.05). Fatty streak lesion size was reduced and the degree of apoptosis was less severe in the KME group (P < 0.05). CONCLUSIONS: In conclusion, antioxidant activity of KME might prevent fatty streak formation through, in part, inhibition of ER stress and apoptosis in aortic sinus where macrophages are harbored.

• 대한한방내과학회지
• /
• 제29권1호
• /
• pp.12-24
• /
• 2008

Background and Objective : Chungsangboha-tang (CSBHT) has analgesic, sedative, anti-convulsive and anti-histamine effects, so it alleviates the symptoms of asthma. For the comparison of anti-inflammatory effect(s) on CSBHT, PD098059 was used as a negative control. Materials and Methods : This study emphasized THP-1 cells, which had been well characterized as a human monocytic leukemic cell line. The cells resemble monocytes with respect to several criteria and can be differentiated into macrophage-like cells by treatment with PMA. By using the MTS assay, it was possible to prove the safety of CSBHT. Results : Results shows that the CSBHT did not affected cell survival within $10^{1}$ ng/ml to $10^{5}$ ng/ml. Especially, $10^{5}$ ng/ml CSBHT treated cells show 70% deduction of $TNF-{\alpha}$ gene expression against that of LPS treated group. Furthermore, $IL-1{\beta}$, IL-6, IL-8, IL-10 and $TNF-{\alpha}$ levels are down-regulated when treated with CSBHT with concentrations up to 100 ug/ml on monocyte-derived macrophages. Interestingly, CSBHT-treated samples showed that overall transcriptional activities were down-regulated to 20% of that of PD098059 ($TNF-{\alpha}$ inhibitor). At protein level, the expression of $TNF-{\alpha}$ showed similar results as that of transcriptional activity. Results show that the protein level decreased more in the CSBHT-treated group (487 ${\pm}$ 87 pg/ml) than in the LPS-treated group (703 ${\pm}$ 103 pg/ml). In addition, the protein level of IL-8 in the CSBHT treated-group (9.84 ${\pm}$ 3.28 ng/ml) decreased similar as the expression of the control and PD098059-treated groups. Conclusion : CSBHT affects immune response, especially allergic responses and suppression of inflammatory reaction. The results provide us an alternative way to care for clinical inflammatory diseases, not only asthma but also the other possible general inflammatory and allergic diseases.