• Title/Summary/Keyword: mouse 3T3-L1

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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.

Effects of IGF-I Rich Fraction from Bovine Colostral Whey on Immune Activity of Mouse In Vitro (젖소 초유로부터 분리한 Insulin-like Growth Factor-1 분획이 In Vitro에서 마우스의 면역 활성에 미치는 영향)

  • Hwang Hyung-A;Yang Hee-Jin;Lee Soo-Won
    • Food Science of Animal Resources
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    • v.26 no.1
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    • pp.113-120
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    • 2006
  • Insulin-like growth factor-I(IGF-I) rich fraction, which was obtained molecules ranged between 30 and 1 kDa, was fractionated by ultrafiltration from bovine colostral whey. IGF-I rich fraction was confirmed by SDS-PACE and western blotting and then the quantity of IGF-I was measured by sandwich ELISA. ICF-I concentration in IGF-I rich fraction was 10 ng/mg proteins. IGF-I rich fraction, standard IGF-I and colostral whey weie treated to murine peritoneal macrophages. And then we experimented that effect of immune activity on macrophage and splenocyte. As a result, in group treated with IGF-I rich fraction $1{mu}g/mL$, production of interleukin-6 and nitric oxide were 9.85 ng and $17.17{\mu}M$ and production of phagocytosis, tumor necrosis factor-${\alpha}\;and\;H_{2}O_{2}$ were 78.3, 34.5 and 6% compared to the control group. In splenocyte immune response, B cell and T cell proliferation and NK cell activity were 103, 126 and 22.2% in group treated with IGF-I rich fraction $1{\mu}g/mL$ to compared to the control, respectively.

Immunomodulating and Antitumor Activities of Panellus serotinus Polysaccharides

  • Kim, Jeong-Hwa;Lee, Jae-Seong;Lee, Kyung-Rim;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyung-Gyun;Cheong, Jong-Chun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • v.40 no.3
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    • pp.181-188
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    • 2012
  • This study was initiated in order to investigate the anticancer and immunomodulating activities of crude polysaccharides extracted in methanol, neutral saline, and hot water (hereinafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of Panellus serotinus. Content of ${\beta}$-glucan and protein in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. serotinus ranged from 22.92~28.52 g/100 g and 3.24~3.68 g/100 g, respectively. In vitro cytotoxicity tests, none of the various fractions of crude polysaccharides were cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at the tested concentration. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 23.53~44.71% in mice previously inoculated with sarcoma 180. Treatment with Fr. HW resulted in an increase in the numbers of spleen cells by 1.3 fold at the concentration of $50{\mu}g/mL$ compared with control. Treatment with Fr. NaCl resulted in improvement of the immuno-potentiating activity of B lymphocytes by increasing the alkaline phosphatase activity by 1.4 fold, compared with control, at the concentration of $200{\mu}g/mL$. Among the three fractions, maximum nitric oxide ($13.48{\mu}M$) was recorded at $500{\mu}g/mL$ in Fr. HW. Production of tumor necrosis factor alpha, interleukin-$1{\beta}$, and interleukin-6 was significantly higher, compared to the positive control, concanavalin A, at the tested concentration. Therefore, treatment with crude polysaccharides extracted from the fruiting body of P. serotinus could result in improvement of antitumor activity.

Induction of Anti-Aquaporin 5 Autoantibody Production by Immunization with a Peptide Derived from the Aquaporin of Prevotella melaninogenica Leads to Reduced Salivary Flow in Mice

  • Ahreum Lee;Duck Kyun Yoo;Yonghee Lee;Sumin Jeon;Suhan Jung;Jinsung Noh;Soyeon Ju;Siwon Hwang;Hong Hee Kim;Sunghoon Kwon;Junho Chung;Youngnim Choi
    • IMMUNE NETWORK
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    • v.21 no.5
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    • pp.34.1-34.16
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    • 2021
  • Sjögren's syndrome (SS) is an autoimmune disease characterized by dryness of the mouth and eyes. The glandular dysfunction in SS involves not only T cell-mediated destruction of the glands but also autoantibodies against the type 3 muscarinic acetylcholine receptor or aquaporin 5 (AQP5) that interfere with the secretion process. Studies on the breakage of tolerance and induction of autoantibodies to these autoantigens could benefit SS patients. To break tolerance, we utilized a PmE-L peptide derived from the AQP5-homologous aquaporin of Prevotella melaninogenica (PmAqp) that contained both a B cell "E" epitope and a T cell epitope. Repeated subcutaneous immunization of C57BL/6 mice with the PmE-L peptide efficiently induced the production of Abs against the "E" epitope of mouse/human AQP5 (AQP5E), and we aimed to characterize the antigen specificity, the sequences of AQP5E-specific B cell receptors, and salivary gland phenotypes of these mice. Sera containing anti-AQP5E IgG not only stained mouse Aqp5 expressed in the submandibular glands but also detected PmApq and PmE-L by immunoblotting, suggesting molecular mimicry. Characterization of the AQP5E-specific autoantibodies selected from the screening of phage display Ab libraries and mapping of the B cell receptor repertoires revealed that the AQP5E-specific B cells acquired the ability to bind to the Ag through cumulative somatic hypermutation. Importantly, animals with anti-AQP5E Abs had decreased salivary flow rates without immune cell infiltration into the salivary glands. This model will be useful for investigating the role of anti-AQP5 autoantibodies in glandular dysfunction in SS and testing new therapeutics targeting autoantibody production.

Effects of BCG on Gastric Chief Cells of the Mouse Implanted with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐 위점막 으뜸세포의 미세구조에 미치는 영향)

  • Ryoo, In-Sang;Ahn, E-Tay;Park, Kyung-Ho;Park, Dae-Kyoon;Kim, Myeong-Soo;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.35 no.3
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    • pp.153-163
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    • 2005
  • This experiment was performed to evaluate the morphological responses of the gastric chief cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG (Bacillus Calmette-Guerin). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with $1x10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the last injection, each mouse was sacrificed. Pieces of the tissue were taken from the stomach, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. The size of zymogen granule and the size of the mitochondrion of the gastric chief cells were observed and calculated. In the BCG treated group, most chief cells did not show any difference in ultrastructure, except that myelin figures were more frequently observed, in comparison with that of nornmal control group. The size of zymogen granule in the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.98({\pm}0.108){\mu}m,\;1.05({\pm}0.092){\mu}m\;and\;0.93({\pm}0.053){\mu}m$, respectively. And the mitochondrial size of the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.80({\pm}0.130){\mu}m,\;0.83({\pm}0.143){\mu}m\;and\;0.72({\pm}0.078){\mu}m$, respectively. From the above results, it was concluded that BCG may slightly suppress function of the gastric chief cells.

Mechanism of Apoptosis & Tumor Growth Inhibition of Agrimonia pilosa Ledebour(APL) in vitro and in vivo (선학초(짚신나물)에 의한 in vitro와 in vivo에서의 암세포사멸 기전 탐색)

  • Choi, Soon-Ja;Baik, Jong-Woo;Park, Jong-Hyeong;Jun, Chan-Yong;Choi, You-Kyung;Ko, Seung-Gyu
    • The Journal of Internal Korean Medicine
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    • v.30 no.2
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    • pp.399-409
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    • 2009
  • Objectives : The aim of this study was to experiment the antitumor activity of Agrimonia pilosa Ledebour (APL) in human stomach cancer (AGS) cell lines (in vitro) and male C57BL/6J mouse (in vivo). Methods : The effects of the ethanol extract from the plant on several transplantable rodent tumors were investigated in vitro by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. DNA content analysis and Western blot analysis. Agrimonia pilosa Ledebour (APL) was given to rats with Lewis Lung Carcinoma (LLC) cells. The experimental rats were divided into 3 groups in vivo. Saline was injected into the abdominal cavity in the first group, 50 mg/kg APL was injected into the abdominal cavity in the second group and 100 mg/kg was injected into the abdominal cavity in the third group. After that, we checked their tumor volume periodically. Results : At first, human gastric cancer (AGS) cell lines (in vitro) showed decreased cell viability, and increased $sub-G_1$ contents. When we experimented rat intestinal epithelial (RIE)l as same condition, this result didn't show. With this, compared to normal cells, Agrimonia pilosa Ledebour (APL) led selectively to the extinction of cells only in human gastric cancer. Moreover, we showed that the traditional herbal medicine APL induced caspase-dependent apoptosis in AGS cells. Next, APL inhibited the growth of LLC-bearing mouse tumor. However, we could not verify APL induced caspase-dependent apoptosis in LLC-bearing mouse tumor. Conclusions : The roots of Agrimonia pilosa Ledebour (APL) contain some antitumor constituents.

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Dead Lactobacillus plantarum Stimulates and Skews Immune Responses toward T helper 1 and 17 Polarizations in RAW 264.7 Cells and Mouse Splenocytes

  • Lee, Hyun Ah;Kim, Hyunung;Lee, Kwang-Won;Park, Kun-Young
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.469-476
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    • 2016
  • This study was undertaken to evaluate the immunomodulatory effect of dead nano-sized Lactobacillus plantarum (nLp) in RAW 264.7 cells and murine primary splenocytes. nLp is a dead, shrunken, processed form of L. plantarum nF1 isolated from kimchi (a traditional Korean fermented cabbage) and is less than 1 μm in size. It was found that nLp treatment stimulated nitric oxide (NO) production more in RAW 264.7 macrophages than pure live L. plantarum (pLp), and that the stimulatory properties were probably largely derived from its cell wall. In addition, nLp induced murine splenocyte proliferation more so than pLp; in particular, a high dose of nLp (1.0 × 1011 CFU/ml) stimulated proliferation as much as lipopolysaccharide at 2 μg/ml. Moreover, according to our cytokine profile results in splenocytes, nLp treatment promoted Th1 (TNF-α, IL-12 p70) responses rather than Th2 (IL-4, IL-5) responses and also increased Th17 (IL-6, IL-17A) responses. Thus, nLp stimulated NO release in RAW 264.7 cells and induced splenocyte proliferation more so than pLp and stimulated Th1 and Th17 cytokine production. These findings suggested that dead nLp has potential use as a functional food ingredient to improve the immune response, and especially as a means of inducing Th1/Th17 immune responses.

Glycosyltransformation of ginsenoside Rh2 into two novel ginsenosides using recombinant glycosyltransferase from Lactobacillus rhamnosus and its in vitro applications

  • Wang, Dan-Dan;Kim, Yeon-Ju;Baek, Nam In;Mathiyalagan, Ramya;Wang, Chao;Jin, Yan;Xu, Xing Yue;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.48-57
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    • 2021
  • Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

Anti-cancer Potentials of Rhus verniciflua Stokes, Ulmus davidiana var. japonica Nakai and Arsenium Sublimatum in Human Gastric Cancer AGS Cells (AGS 인체위암세포에서 건칠, 유근피 및 신석 추출물의 항암 활성 비교 연구)

  • Baek, Ilsung;Im, Lyeng-Hae;Park, Cheol;Cho, Yung Hyun
    • Journal of Life Science
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    • v.25 no.8
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    • pp.849-860
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    • 2015
  • The anti-cancer activities of Rhus verniciflua Stokes (GC), Ulmus davidiana var. japonica Nakai (UGP) and arsenium sublimatum (SS) extracts, which have been used Oriental medicine therapy for various diseases, were investigated. The treatment of GC, UGP and SS alone, and combined treatment with GC, UGP and SS did not affect the cell viability in the mouse normal cell lines (RAW 264.7 macrophages and C2C12 myoblasts). However, co-treatment with GC, UGP and SS markedly induces apoptosis in human gastric cancer AGS cells, but not in other various cancer cell lines (human lung cancer A549, colon cancer HCT116, liver cancer Hep3B and bladder T24 cells) as evidenced by formation of apoptotic bodies, chromatin condensation, and accumulation of annexin-V positive cells. Co-treatment with GC, UGP and SS effectively induced the expression levels of Fas and Fas ligand, and inhibited the levels IAP family proteins such as XIAP, cIAP-1 and survivin, and anti-apoptotic Bcl-xL proteins compared with treatment with either agent alone. Combined treatment also significantly induced the loss of mitochondrial membrane potential, which was associated with the activation of caspases (-3, -8, and -9) and degradation of poly (ADP-ribose) polymerase. However, the cytotoxic effects induced by co-treatment with GC, UGP and SS were significantly attenuated by pan-caspases inhibitor, z-VAD-fmk, indicating an important role for caspases. These results indicated that the caspases were key regulators of apoptosis in response to co-treatment of GC, UGP and SS in human gastric cancer AGS cells and further studies will be needed to identify the active compounds.

Immunohistochemical Study on the Activation of Cell mediated immunity in Murine Lymph node on Allergic Contact Dermatitis by DNCB -Based on the change of T lymphocytes and Il-2 receptors- (알러지성 접촉피부염 유발 피부 주변 림프절에서의 세포성 면역 활성에 관한 면역조직화학적 연구 - T 림프구와 IL-2 수용기의 분포 변화를 중심으로 -)

  • kim, Jin-Taek;Ahn, Sang-Hyun;Park, In-Sick;Chung, Jae-Man;Kim, Ho-Hyun
    • The Journal of Dong Guk Oriental Medicine
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    • v.7 no.1
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    • pp.33-41
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    • 1998
  • Lymph node tissues of BALB/C mouse treated with DNCB were immunohistochemically observed to investigate the activation of cell mediated immunity in lymph node of murine with allergic contact dermatitis. The inguinal region of BALB/C mice were sensitized by one application of $25{\mu}l$ of 5% 2,4-dinitrochlorobenzene(DNCB) onto an abdominal skin and 2 weeks later, the mice were challenged with $4{\mu}l$ of 2.5% DNCB. The inguinal lymph node were obtained at hour 24, 48, and 72 after 2nd DNCB treatment and embedded with paraffin, and then stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly2(CD8), IL-2R(CD25). The distribution of helper T lymphocytes, cytotoxic T lymphocytes and IL-2 receptors began to increase at hour 24 after after 2nd DNCB treatment and these increase appeared in paracortical area and medullary sinius. These increase were greatest at hour 48. These results indicated that the IL-2 secretion began to increase by activation of helper T lymphocytes in lymph node of DNCB re-exposure area and subsequently to activate suppress T lymphocytes.

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