• Title/Summary/Keyword: mouse B cell

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Inhibitory Effect of Rhizoma Bletillae on Melanogenesis of B16 Melanoma Cell (白급이 B16 흑색종세포의 멜라닌 형성 억제에 미치는 영향)

  • Yoon, Hwa-jung;Yoon, Jung-won;Yoon, So-won;Ko, Woo-shin;Woo, Won-hong
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.16 no.3
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    • pp.129-144
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    • 2003
  • Recently many efforts were focused to understand the mechanical insights of melanogenesis to develop the agents for hyper-pigmentation and hypo-pigmentation. In the melanin biosynthetic pathway, tyrosinase is the rate limiting enzyme, and ${\alpha}$-melanocyte stimulating hormone(MSH) or cAMP-elevating agents stimulate melanogenesis and enhance the melanin synthesis and the tyrosinase activity. The author has analyzed the effects of Rhizoma Bletillae on the basal melanogenic activities of B16/F10 mouse melanoma cells, and on the ${\alpha}$-MSH or forskolin-induced melanogenesis. Rhizoma Bletillae alone markedly suppressed melanin content and tyrosinase activity in a dose-dependent manner. Pretreatment of the cells with Rhizoma Bletillae also suppressed the increase of ${\alpha}$-MSH (100 nM) or forskolin (20 ${\mu}M$)-induced melanin content and tyrosinase activity. The decrease in the tyrosinase activity was paralled by a decrease in the abundance of tyrosinase protein and tyrosinase promoter activity. Pretreatment of the cells with Rhizoma Bletillae also inhibited the increase of forskolin(20${\mu}M$) induced the amount of tyrosinase protein and tyrosinase promoter activity. The results of DOPA staining revealed that pretreatment of the cells with Rhizoma Bletillae showed less intensity than B16 melanoma cells stimulated with ${\alpha}$-MSH or forskolin. These results suggest that Rhizoma Bletillae inhibits melanogenesis and abrogates ${\alpha}$-MSH and cAMP-induced melanogenesis in B16 melanoma cells.

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Action Mechanism of LB10522, a New Catechol-Substituted Cephalosporin (카테콜 치환체를 가진 세파로스포린계 항생제 LB10522의 작용기전)

  • Kim, Mu-Yong;Oh, Jeong-In;Paek, Kyoung-Sook;Kim, In-Chull;Kwak, Jin-Hwan
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.102-111
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    • 1996
  • LB10522 is a new parenteral broad spectrum cephalosporin with a catechol moiety at C-7 position of beta-lactam ring. This compound can utilize tonB-dependent iron transp ort system in addition to porin proteins to enter bacterial periplasmic space and access to penicillin-binding proteins (PBPs) which are the lethal targets of ${\beta}$-lactam antibiotics. The chelating activity of LB10522 to metal iron was measured by spectrophotometrically scanning the absorbance from 200 to 900nm. When $FeCl_3$ was added, optical density was increased between 450 and 800nm. LB10522 was more active against gram-negative strains in iron-depleted media than in iron-replete media. This is due to the increased expression of iron transport channels in iron-depleted condition. LB10522 showed a similar activity against E. coli DC2 (permeability mutant) and E. coli DCO (wild type strain) in both iron-depleted and iron-replete media, indicating a minimal permeaility barrier for LB10522 uptake. LB10522 had high affinities to PBP 3 and PBP 1A, 1B of E. coli. By blocking these proteins, LB10522 caused inhibition of cell division and the eventual death of cells. This result was correlated well with the morphological changes in E. coli exposed to LB10522. Although the in vitro MIC of LB10522 against P. aeruginosa 1912E mutant (tonB) was 8-times higher than that of the P. aeruginosa 1912E parent strain, LB10522 showed a similar in vivo protection efficacy against both strains in the mouse systemic infection model. This result suggested that tonB mutant, which requires a high level of iron for normal growth, might have a difficulty in surviving in their host with an iron-limited environment.

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Cadmium-induced COX-2 Expression in Cerebrovascular Endothelial Cells (카드뮴이 뇌혈관 내피세포에서의 $PGE^2$ 및 COX-2 발현에 미치는 영향)

  • Park Dong-Hyun;Kim Young-Chae;Moon Chang-Kiu;Jung Yi-Sook;Baik Eun-Joo;Moon Chang-Hyun;Lee Soo-Hwan
    • Environmental Analysis Health and Toxicology
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    • v.21 no.3 s.54
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    • pp.275-282
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    • 2006
  • In order to get insight into the mechanism of cadmium (Cd)-induced brain injury, we investigated the effects of Cd on the induction of COX-2 in bEnd.3 mouse brain endothelial cells. Cd induced COX-2 expression and $PGE_2$ release, which were attenuated by thiol-reducing antioxidant N-acetylcysteine (NAC) indicating oxidative components might contribute to these events. Indeed, Cd increased cellular reactive oxygen species (ROS) level and DNA binding activity of nuclear factor-kB (NF-kB), an oxidative stress sensitive transcription factor. Cd-induced $PGE_2$ production and COX-2 expression were significantly attenuated by Bay 11 7082, a specific inhibitor of NF-kB and by SB203580, a specific inhibitor of p38 mitogen activated protein kinase (MAPK). These data suggest that Cd induces COX-2 expression through activation of NF-kB and p38 MAPK, the oxidative stress-sensitive signaling molecules, in brain endothelial cells.

Expression Analysis of the Ligand to Ly-6E.1 Mouse Hematopoietic Stem Cell Antigen

  • Hwang, Dae-Youn;Min, Dul-Lei;Sonn, Chung-Hee;Chang, Mi-Ra;Lee, Mi-Hyun;Paik, Sang-Gi;Kim, Young-Sang
    • Animal cells and systems
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    • v.1 no.1
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    • pp.157-164
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    • 1997
  • Ly-6E.1 antigen was proposed as a regulatory molecule of T lymphocyte activation, a hematopoietic stem cell marker, a memory cell marker, and an adhesion molecule. Though there were several reports suggesting the presence of Ly-6 ligand, the characterization of the ligand was not yet performed, As an attempt to screen the expression of Ly-6E.1 ligand, we prepared a probe for detecting Ly-6E.1 ligand by producing a fusion protein between Ly-6E.1 and $hlgC_{r1}$, A mammalian cell expression vector with Ly-6E.$1/hlgC_{r1}$ chimeric cDNA was transfected in SP2/0-Ag14 myeloma cells, and stable transfectants were selected. The fusion protein was produced as a dimer and maintained the epitopes for monoclonal antibodies specific for Ly-6E.1 and for anti-human lgG antibody. The purified fusion protein through Gammabind G column was used for FACS analyses for the expression of Ly-6E.1 ligand. The fusion protein interacted with several cell lines originating from B cells, T cells, or monocytes. The fusion Protein also strongly stained bone marrow, lymph node, and spleen cells, but thymic cells weakly, if any. The staining was more obvious in C57BL/6 $(Ly-6^b)$ than Balb/c $(Ly-6^a)$ mice. These results suggest that the interaction of Ly-6E.1 with Ly-6E.1 ligand may function both in the stem cell environment and in the activation of mature lymphocytes. The fusion protein may be a valuable tool in characterization of biochemical properties of the Ly-6E.1 ligand and, further, in isolating its cDNA.

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UVB-Shielding Effects of para-Coumaric Acid (파라-쿠마린산의 자외선B 차단 효과)

  • Song, Kyo-Sun;Boo, Yong-Chool
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.3
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    • pp.263-273
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    • 2012
  • Recent studies have uncovered attractive properties of para-coumaric acid (PCA) as a potential skin hywhitening agent. The purpose of the current study was to examine its UVB-shielding effects. Effects of PCA on the viability of HaCaT cells exposed to UVB were assessed in vitro in comparison with other aromatic amino acid metabolites that have similar UV absorption spectra. For in vivo test, PCA cream (1.5 %) and cream base were topically applied to the dorsal skin of SKH-1 hairless mice and the inflammatory responses due to UVB exposure were monitored by changes in skin color (erythema) and thickness (edema). The cream application-UVB exposure regimen was repeated every other day for a total of 12 sessions. When HaCaT cells were irradiated with UVB, there was a dose-dependent decline in cell viability. The cell viability decline due to UVB exposure (10 mJ $cm^{-2}$) was significantly prevented by 100 ${\mu}M$ PCA, cinnamic acid, urocanic acid, or indole acrylic acid by 39, 27, 39, or 31 %, respectively. Topical application of PCA cream onto the dorsal skin of hairless mice (10 ${\mu}g\;cm^{-2}$) attenuated the changes of color parameters, $L^*$, $a^*$, $b^*$ values, and thickness of the UVB (150 mJ $cm^{-2}$)-exposed skin by 59, 50, 58, and 53 %, respectively. The current study, together with the previous studies that demonstrated the antimelanogenic effects of PCA, suggested that PCA may prevent not only dyspigmentation but also inflammatory reactions in the UVB-exposed skin.

Suppressive Effect of Kwanjulbang-6 on Collagen-Induced Arthritis in Mice (관절6호방이 Type II collagen으로 유발된 관절염에 미치는 영향)

  • Kim, Hyun-Soo;Hong, Seo-Young;Heo, Dong-Seok;Oh, Min-Suck
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.5
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    • pp.1003-1011
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    • 2009
  • This study was carried out to know the effects of Kwanjulbang-6(GJB-6) on the inhibition of arthritis. GJB-6 was orally administered to mouse with arthritis induced by collagen II . Cytotoxicity, hepatotoxicity, arthritis index, value of immunocyte in draining lymph node and paw joint, rheumatoid factor in serum were measured in vivo. The incidence of arthritis was significantly decreased. Total cell number of draining lymph node was significantly increased compared with control. Total cell number of paw joint was significantly decreased compared with control. The absolute number of $CD19^+$, $CD8^+$, $CD3^+/CD69^+$, $CD3^+/CD49b^+$, $CD4^+/CD44^+$, $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD3^+/CD8^+$ cells in draining lymph node were significantly increased compared with control. The absolute number of $CD3^+$, $CD4^+$, $CD4^+/CD25^+$ and $CD11b^+/Gr-1^+$ cells in paw joint were significantly decreased compared with control. The absolute number of $B220^+/CD23^+$ and $MHCII^+/CD11c^+$ cell in draining lymph node were significantly decreased compared with control. The levels of IgG was decreased and The levels of IgM was significantly decreased compared with control. Anti-collagen II in serum was significantly decreased compared with control. With the hematoxylin and eosin stain, the cartilage destruction and synovial cell proliferation were decreased compared with control. With the Masson's trichrome stain, the expression of collagen fibers was decreased compared with control. Results showed that GJB-6 had immunomodulatory effects. So we expect that GJB-6 should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease.

산삼의 배양 및 그 응용에 관한 연구

  • 신미희
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.2
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    • pp.45-56
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    • 2001
  • Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extract. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were Patch teated in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively.

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Antitumor and Immuno-potentiating Activity against Mouse Sarcoma 180 by Crude Polysaccharides Extracted from Fruiting Body of Tricholoma matsutake (송이(Tricholoma matsutake)의 자실체에서 추출한 조다당류가 생쥐의 Sarcoma 180에 미치는 항암 및 면역증강 작용)

  • Hur, Hyun;Choi, Yon-Il;Lee, Tae-Soo
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1290-1298
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    • 2008
  • Tricholoma matsutake, one of edible and medicinal mushroom belonging to Tricholomaceae of Agaricales, has been known to contain some curing effect on gastric cancer and ulcer, and inhibitory effect on sarcoma 180 and Ehrlich sarcoma. Neutral salt soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from fruiting body of the mushroom. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HepG2, HT29 and NIH3T3 at the concentration of 2.0 mg/ml. Intraperitoneal injection with crude polysaccharides showed life prolongation effect of 23.4$\sim$37.2% in mice previously inoculated with Sarcoma 180. Fr. MeOH and Fr. HW exhibited the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 2.2$\sim$11.9 folds compared with control at the concentration of 0.2$\sim$0.5 mg/ml. In case of Fr. NaCl, the numbers of peritoneal exudate cells and circulating leukocytes were increased by 6.0 and 1.5 folds at the concentration of 50 mg/kg, respectively. Therefore, it is concluded that crude polysaccharides extracted from fruiting body of Tricholoma matsutake showed antitumor and immuno-potentiating activity against Sarcoma 180 of mouse.

Antioxidant Activity and Radioprotection of Two Flavonoids from Propolis (프로폴리스에서 분리한 플라보노이드 화합물의 항산화 활성 및 방사선 방어효과)

  • Jeong, Ill-Yun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.162-166
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    • 2005
  • Two flavonoids, 7-O-methyl-3',4'-didehydroxy quercetin (MDQ) and quercetin, isolated from Chinese propolis, which is the generic name for the resinous substance collected by honeybees from various plant sources, were tested for their antioxidant activity and protective effect against radiation-induced DNA damage in mouse lymphocytes. In antioxidant test, both compounds provided a dose-dependent scavenging effect on DPPH radical and a dose-dependent inhibitory effect on lipid peroxidation in mouse liver. Quercetin showed stronger scavenging and inhibitory effect than MDQ, and it also provided strong inhibition on superoxide anion radical generated in xanthine-xanthine oxidase system, but there was no inhibitory ability for MDQ. In comet assay using single cell gel electrophoresis, MDQ and quercetin showed a protective effect against DNA damage caused by gamma irradiation. They reduced DNA damage to 54% (p<0.01) and 53% (p<0.01) at 25 $\mu$mol, respectively. These results suggest that free radical scavenging seems to be associated with their catechol form on the B ring, and radioprotection appears to be a likely mechanism of antioxidant activity by these flavonoids.

Immunization of Mice with Recombinant Brucella abortus Organic Hydroperoxide Resistance (Ohr) Protein Protects Against a Virulent Brucella abortus 544 Infection

  • Hop, Huynh Tan;Reyes, Alisha Wehdnesday Bernardo;Simborio, Hannah Leah Tadeja;Arayan, Lauren Togonon;Min, Won Gi;Lee, Hu Jang;Lee, Jin Ju;Chang, Hong Hee;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.190-196
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    • 2016
  • In this study, the Brucella abortus ohr gene coding for an organic hydroperoxide resistance protein (Ohr) was cloned into a maltose fusion protein expression system (pMAL), inserted into Escherichia coli, and purified, and its immunogenicity was evaluated by western blot analysis using Brucella-positive mouse sera. The purified recombinant Ohr (rOhr) was treated with adjuvant and injected intraperitoneally into BALB/c mice. A protective immune response analysis revealed that rOhr induced a significant increase in both the IgG1 and IgG2a titers, and IgG2a reached a higher level than IgG1 after the second and third immunizations. Additionally, immunization with rOhr induced high production of IFN-γ as well as proinflammatory cytokines such as TNF, MCP-1, IL-12p70, and IL-6, but a lesser amount of IL-10, suggesting that rOhr predominantly elicited a cell-mediated immune response. In addition, immunization with rOhr caused a significantly higher degree of protection against a virulent B. abortus infection compared with a positive control group consisting of mice immunized with maltose-binding protein. These findings showed that B. abortus rOhr was able to induce both humoral and cell-mediated immunity in mice, which suggested that this recombinant protein could be a potential vaccine candidate for animal brucellosis.