• Title/Summary/Keyword: Hexosaminidase

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Inhibitory effect of Ulmus davidiana on ${\beta}$-hexosaminidase release and cytokine production in RBL-2H3 cells (유근피 추출물이 RBL-2H3 비만세포에서 ${\beta}$-hexosaminidase 및 cytokine 분비에 미치는 효과)

  • Park, Se-Bong;Kang, Kyung-Hwa;Yoon, Hwa-Jung;Ko, Woo-Shin
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.24 no.1
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    • pp.86-95
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    • 2011
  • Objectives : Ulmus davidiana (UD) has been widely used in Korean herbal medicines used for treatment of acute and chronic inflammatory diseases, such as rhinitis, asthma, and abscess. In this study, To investigated the protective effect of UD on type 1 allergic response, we determined whether UD inhibits early and late allergic response. Methods : The effect of UD was analyzed by ELISA and RT-PCR in RBL-2H3 cells. Levels of ${\beta}$ -hexosaminidase, interleukin (IL)-4 and TNF-${\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISAs). mRNA levels of COX-2 and T-helper type 2(Th2) cytokines were analyzed with RT-PCR. Results : We found that UD suppressed ${\beta}$-hexosaminidase release in RBL-2H3 not only by the PMA plus A23187 stimulation, but also by the IgE-DNP-HSA stimulation at the antigen-antibody binding stage and antibody-receptor binding stage. UD also significantly inhibited COX2 level, along with reduced Th2 cytokine levels, such as IL-3, IL-4, IL-5, IL-13, GM-CSF, and TNF-${\alpha}$ in RBL-2H3. Conclusions : Our results indicate that UD protects against type 1 allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and expression of COX2 and Th2 cytokines.

Intracellular $Ca^{2+}$ Mobilization and Beta-hexosaminidase Release Are Not Influenced by 60 Hz-electromagnetic Fields (EMF) in RBL 2H3 Cells

  • Hwang, Yeon-Hee;Song, Ho-Sun;Kim, Hee-Rae;Ko, Myoung-Soo;Jeong, Jae-Min;Kim, Yong-Ho;Ryu, Jeong-Soo;Sohn, Uy-Dong;Gimm, Yoon-Myoung;Myung, Sung-Ho;Sim, Sang-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.5
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    • pp.313-317
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    • 2011
  • The effects of extremely low frequency electromagnetic fields (EMF) on intracellular $Ca^{2+}$ mobilization and cellular function in RBL 2H3 cells were investigated. Exposure to EMF (60 Hz, 0.1 or 1 mT) for 4 or 16 h did not produce any cytotoxic effects in RBL 2H3 cells. Melittin, ionomycin and thapsigargin each dose-dependently increased the intracellular $Ca^{2+}$ concentration. The increase of intracellular $Ca^{2+}$ induced by these three agents was not affected by exposure to EMF (60 Hz, 1 mT) for 4 or 16 h in RBL 2H3 cells. To investigate the effect of EMF on exocytosis, we measured beta-hexosaminidase release in RBL 2H3 cells. Basal release of beta-hexosaminidase was $12.3{\pm}2.3%$ in RBL 2H3 cells. Exposure to EMF (60 Hz, 0.1 or 1 mT) for 4 or 16 h did not affect the basal or $1{\mu}m$ melittin-induced beta-hexosaminidase release in RBL 2H3 cells. This study suggests that exposure to EMF (60 Hz, 0.1 or 1 mT), which is the limit of occupational exposure, has no influence on intracellular $Ca^{2+}$ mobilization and cellular function in RBL 2H3 cells.

Inhibitory Effects of Scrophulariae Radix on β-hexosaminidase release and cytokine production in RBL-2H3 cells (현삼(玄蔘) 추출물이 RBL-2H3 비만세포에서 β-hexosaminidase 및 cytokine 분비에 미치는 효과)

  • Kim, Se-Gie
    • The Korea Journal of Herbology
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    • v.32 no.6
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    • pp.9-15
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    • 2017
  • Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.

Fractions of Chamaecyparis obtusa Display Antiallergic Effect in RBL2H3 Cells

  • Choi, In-Gyu;Kim, Kyung-Jong;Kim, Young-Mi;Park, Mi-Jin;Lee, Yun-Sil;Jeoung, Doo-Il
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1747-1752
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    • 2006
  • Allergic inflammation results from stimulation of ${\beta}$-hexosaminidase secretion, increased calcium influx, and activation of MAPK pathways. Some fractions of Chamaecyparis obtusa decreased secretion of ${\beta}$-hexosaminidase, calcium influx, ROS, and phosphorylation of ERK. These results suggest that Chamaecyparis obtusa would be valuable for development of allergy therapeutics.

Anti-allergic Effects of Cheonmaec-tang in RBL-2H3 Cell (천맥탕(天麥湯)이 항알러지 염증반응에 미치는 영향)

  • Kim, Yong-Hyeon;Lee, Seung-Yeon;Kim, Won-Il
    • The Journal of Pediatrics of Korean Medicine
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    • v.22 no.1
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    • pp.95-102
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    • 2008
  • Objectives The purpose of this study was to examine the anti allergic reaction with Cheonmaec-tang. Methods We examined Cell Viability, ${\beta}$-hexosaminidase, TNF-${\alpha}$, IL-4 secretion from RBL-2H3 cell after pretreatment with 2 ㎎/ml, 4 ㎎/ml of Cheonmaec-tang. Results We observed that Cheonmaec-tang is reduced to ${\beta}$-hexosaminidase, TNF-${\alpha}$, IL-4 secretion in RBL-2H3 cell. Conclusions These results indicate that Cheonmaec-tang has anti-histamic effect and controls TNF-${\alpha}$, IL-4 secretion on allergic reaction.

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Inhibitory Effect of Extracts from Rhododendron Brachycarpum and Abies Koreana E.H. Wilson on Degranulation and Cytokine Expression in RBL-2H3 Cells (만병초와 구상나무 추출물의 RBL-2H3 세포 탈과립, 싸이토카인 유전자 발현에 미치는 영향)

  • Jung, Eui-Man;Kim, Jae-Woo;Park, Mi-Jin;Lee, Sung-Suk;Choi, Don-Ha;Lee, Jungbok;Jeung, Eui-Bae
    • Journal of the Korean Wood Science and Technology
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    • v.41 no.6
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    • pp.551-558
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    • 2013
  • Ethanol extracts from Rhododendrom brachycarpum and Abies koreana were investigated for their anti-allergic activities using RBL-2H3 cell line. After treatment with ethanol extracts of various concentrations on the immune response induced mast cell by concanavalin A (Con A), the expressions of cytokine interleukin-4 (IL-4), interleukin-13 (IL-13) were determined by using RT-PCR and the degranulation of mast cells was determined by measuring ${\beta}$-hexosaminidase release. Expression level of IL-4 was decreased by the extract from Rhododendrom brachycarpum in $10^{-7}$, $10^{-5}$ and $10^{-3}%$ concentrations. Expression level of IL-13 was also decreased by both extracts. ${\beta}$-Hexosaminidase release by RBL-2H3 cells was inhibited at the $10^{-5}$ and $10^{-3}%$ concentration of extracts from Rhododendrom brachycarpum and Abies koreana, respectively. These results demonstrate that ethanol extracts of Rhododendron brachycarpum and Abies koreana exert anti-allergic effects by regulating the reduction of IL-4 and IL-13 genes expression and also the secretion of ${\beta}$-hexosaminidase.

Comparison of Anti-allergenic Activities of Various Polyphenols in Cell Assays

  • Yun, Sang-Sik;Kang, Mi-Young;Park, Jun-Cheol;Nam, Seok-Hyun
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.139-146
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    • 2010
  • The inhibitory effects of 25 polyphenols against in vitro allergic reactions were compared using biochemical and cell assays. Three polyphenols including curcumin, gallic acid, and quercetin suppressed the release of $\beta$-hexosaminidase from ionophore A23187-stimulated RBL-2H3 cells more effectively (>50% inhibition at $100{\mu}M$ concentration). They were found to have potencies in suppressing the release of histamine not only from ionophore A23187-, but also from immunoglobulin E (IgE)-stimulated RBL-2H3 cells. Moreover, such suppressive effects of the three polyphenols were also observed in A23187 plus PMA-costimulated rat peritoneal mast cells. The extent of inhibition were quantified as the respective polyphenol concentration that inhibit 50% ($IC_{50}$) of $\beta$-hexosaminidase or histamine release, showing an inhibition tendency with decreasing order of curcumin>gallic acid>quercetin. Down-regulation of $Ca^{2+}$ influx was suggested as the cause of the inhibition of $\beta$-hexosaminidase and histamine releases in these cells. The immune process inhibition was confirmed by the observed reduction in the gene expressions and release of pro-inflammatory cytokine tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-$1\beta$, and IL-4, due probably to antioxidant activity of the polyphenols. These findings illustrate that curcumin, gallic acid, and quercetin may be beneficial against allergic inflammatory diseases.

Inhibitory Effects of Punica granatum L. Extracts on Degranulation in Human Basophilic KU812F Cells (석류 추출물에 의한 인간호염구(KU812F 세포)의 탈과립 억제효과)

  • Park, Kyong-Tae;Shim, Sun-Yup;Chun, Soon-Sil
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.702-706
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    • 2008
  • Punica granatum (PG) evidences a variety of physiological properties, including anti-diabetic, anti-cancer, antiinflammatory, anti-microbial, and anti-oxidative activities. Using the human basophilic KU812F cells, the inhibitory effects of the methanolic extract of PG seed, shell, and juice on calcium ionophore, A23187-induced degranulation were assessed. All of the PG extracts inhibited A23187-induced intracellular $Ca^{2+}$ levels, ${\beta}$-hexosaminidase, and histamine release in a dose-dependent manner. These results showed that all of the PG extracts are potent inhibitors of degranulation in allergic reactions, via the suppression of $Ca^{2+}$ influx.

Inhibitory Effects of Soyasaponins on Antigen-induced Degranulation in RBL-2H3 Cells (Soyasaponin의 RBL-2H3 세포에서 탈과립화 억제 효과)

  • Yang, Seung Hwan;Lee, Jung A;Lee, Jae Yeon;Ahn, Eun-Kyung;Shin, Tai-Sun;Tsukamoto, Chigen;Chung, Gyuhwa;Suh, Joo-Won
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.287-290
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    • 2015
  • Soyasaponins are glycosylated, which gives rise to a wide diversity of structures and functions. We evaluated for inhibitory effects of 4 soyasaponins on antigen-induced degranulation in RBL-2H3 Cells. 4 soyasaponins had shown dose-dependently inhibited histamine and ${\beta}$-hexosaminidase.

Inhibition Activity of Plants on IgE-mediated Degranulation of RBL-2H3 Cells

  • Lee, Seung-Eun;Jeong, Hye-Gwang;Lee, Dae-Young;Lee, Jeong-Hoon;Choi, Jehun;Kim, Geum-Soog;Noh, Hyung-Jun;Lee, Jae-Won;Kim, Seung-Yu;Ahn, Young-Sup
    • Korean Journal of Plant Resources
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    • v.28 no.6
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    • pp.718-726
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    • 2015
  • In this study, the inhibitory activities of fifty plant extracts on IgE-mediated degranulation in the rat basophilic leukemia cell line (RBL-2H3 cells) were measured; the release of interleukin (IL)-4 and β-hexosaminidase from IgE-sensitized cells treated with the plant extracts was measured; and the effects of the plant extracts on cell viability were tested. The results of the analysis of plant extracts at 20 μg/ml, including the aerial part of Magnolia sieboldii K. Koch, exhibited suppressive activities upon the release of IL-4. Furthermore, several plant extracts including methanol extracted from Lindera erythrocarpa Makino (aerial part) at the same concentration significantly inhibited the release of β-hexosaminidase. Twenty-six of the plant extracts, including methanol extract of Weigela subsessilis (Nakai) L. H. Bailey (branch), showed a cell proliferation effect of over 80% at 100 μg/ml. In conclusion, the results suggest that the leaf/stem of Geum japonicum Thunb. and the stamen/ovary of Nelumbo nucifera Gaertn., which exhibited effective inhibition on β-hexosaminidase release and IL-4 release from mast cells and showed high cell viability, could be useful candidates as anti-allergy materials.