• Title/Summary/Keyword: Beta-hexosaminidase

Search Result 119, Processing Time 0.018 seconds

Effect of modified-Okbyungpoongsan on mast cell-mediated allergic responses in RBL-2H3 mast cells (가미옥병풍산(加味玉屛風散)의 비만세포에서의 알레르기 반응에 대한 효과 연구)

  • Jung, Jin-Ki;Park, Yong-Ki
    • The Korea Journal of Herbology
    • /
    • v.26 no.4
    • /
    • pp.1-7
    • /
    • 2011
  • Objectives : In this study, we investigated the effect of modified-Okbyungpoongsan (mOP) on mast cell-mediated allergic response in basophilic leukemia cell line, RBL-2H3 mast cells. Methods : Cells were stimulated with anti-DNP-IgE after the treatment of DNP-HSA (AI/D), and then incubated with different concentrations of mOP (0.1, 0.2, 0.5 and 1 mg/$m{\ell}$) in RBL-2H3 cells. Cell toxicity was determined by WST-1 assay. The degranulation of mast cells was observed by microscope with toluidine blue staining and also the levels of beta-hexosaminidase, histamine and TNF-alpha were measured in culture supernatants by enzyme-linked immunosorbant assay. Results : mOP inhibited anti-DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. mOP also significantly decreased the levels of histamine and inflammatory cytokine, TNF-alpha in RBL-2H3 cells, but slightly decreased the level of beta-hexosaminidase. Conclusions : These results indicate that mOP, an oriental prescription could be inhibit the allergic response through suppressing the mast cell activation.

Suppressive Effects of Fucoxanthin on Degranulation in IgE-antigen complex-stimulated RBL-2H3 Cells

  • NamKoong, Seung;Joo, Hae-Mi;Jang, Seon-A;Kim, Ye-Jin;Kim, Tae-Seong;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
    • /
    • v.25 no.3
    • /
    • pp.339-345
    • /
    • 2012
  • The marine carotenoid fucoxanthin can be found in marine brown seaweeds, macroalgae, diatoms, and microalgae, and has remarkable biological properties. Numerous studies have shown that fucoxanthin has considerable potential and promising applications in human health, but the underlying mechanisms involved in its anti-allergic activity are not fully understood. We here investigated the mechanisms by anti-allergic activity of fucoxanthin fraction from Eisenia bicyclis in immunoglobulin E-antigen complex (IgE/DNP-BSA)-stimulated RBL-2H3 mast cells. This study we found that the fucoxanthin inhibits the release of ${\beta}$-hexosaminidase and suppressed not only transcriptional activation of NF-${\kappa}B$, but also phosphorylation of ERK and JNK in IgE/DNP-BSA-treated RBL-2H3 cells. Fucoxanthin may be useful for preventing allergic diseases, including asthma and atopic dermatitis.

Effects of Hyeongbangpaedok-san (HBPDS) on Allergic Contact Dermatitis (ACD) induced by DNFB in Mice (형방패독산(荊防敗毒散)이 DNFB로 유발된 생쥐의 알레르기접촉피부염에 미치는 영향)

  • Park, Geun-Hee;Lim, Tae-Hyung;Park, Hye-Sun
    • Journal of Sasang Constitutional Medicine
    • /
    • v.26 no.2
    • /
    • pp.180-193
    • /
    • 2014
  • Objectives The present study was carried out to investigate effects of HBPDS on allergic contact dermatitis (ACD) induced by 2,4-Dinitro-1-fluorobenzene (DNFB) in mice. Methods In this experiment, effects of HBPDS on body weights, skin thicknesses, skin weights, histopathological changes, clinical aspects, erythema index, melanin index, production levels of cytokines in ACD mice were investigated. In addition, effects on proliferation rates, release of ${\beta}$-hexosaminidase and histamine were also investigated in vitro. Results & Conclusions 1) HBPDS inhibited enlargement of skin thickness and weight significantly (P < 0.05). 2) HBPDS treatment prevented spongiosis, edema and immune cell infiltrations. 3) Erythema, desquamation and keratosis were diminished by oral administration of HBPDS. 4) Production levels of TNF-alpha and IFN-gamma in serum were decreased by HBPDS treatment in vivo. 5) More than 200 ${\mu}g/ml$ of HBPDS treatment decreased ${\beta}$-hexosaminidase release and more than 400 ${\mu}g/ml$ of HBPDS treatment also decreased histamine release in vitro.

Anti-allergic Activity of Dojuk-San Ethanol Extract (도적산(導赤散) 에탄올 추출물의 알레르기 억제효과)

  • Sung, Sun-Heui;Lee, Su-Kyoung;Kang, Ok-Hwa;Kwon, Dong-Yeul;Chong, Myong-Soo;Lee, Ki-Nam
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.25 no.3
    • /
    • pp.438-444
    • /
    • 2011
  • Dojuk-San is known to be effective for treating a urinary diseases and stomatitis. However, its effects on the bone marrow-derived mast cell(BMMC) mediated allergy and inflammation mechanism remain unknown. In this study, the biological effects of Dojuk-San ethanol extract(DJS) were evaluated while focusing on its effects on the allergic mediator in phorbol 12-myristate 13-acetate(PMA) plus calcium ionophore A23187(A23187)-stimulated BMMCs. An investigation was also conducted to determine its effects on the production of several allergic mediators including interleukin-6(IL-6), prostaglandin D2($PGD_2$), leukotrieneC4(LTC4) and ${\beta}$-Hexosaminidase(${\beta}$-Hex). The results revealed that DJS inhibited the PMA plus A23187 induced production of IL-6, PGD2, LTC4 and ${\beta}$-Hex. Taken together, these findings indicate that DJS has the potential using in the treatment of allergy.

The comparative study of anti-allergic and anti-inflammatory effects by fermented red ginseng and red ginseng (홍삼과 발효홍삼의 항염증 작용 및 항알러지 효과 비교)

  • Park, Hye-Jin;Jung, Da-Hye;Joo, Hae-Mi;Kang, Nam-Sung;Jang, Seon-A;Lee, Jae-Geun;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
    • /
    • v.23 no.5
    • /
    • pp.415-422
    • /
    • 2010
  • Red ginseng(RG, steamed and dried root of Panax ginseng C. A. Meyer, family Araliaceae) and fermented red ginseng(FRG, fermented red ginseng by yeast and lactic acid bacteria) are known to show different pharmacological effects by changed composition of saponins through fermentation. We examined the effects of RG and FRG on $\beta$-hexosaminidase secretion, ICAM-1 expression, the mitogen-induced proliferation of lymphocyte from mice in ex vivo systems and HaCaT cell(keratinocyte) proliferation to compare the anti-allergic and anti-inflammatory effects between both groups. RG groups showed inhibition of $\beta$-hexosaminidase secretion and ICAM-1 expression at $1{\mu}g/ml$, $10{\mu}g/ml$ and the same effects were observed at all concentrations in FRG groups. In our study, RG increased LPS-induced B cell proliferation at $1{\mu}g/ml$ and ConA-induced B cell proliferation at $100\;{\mu}g/ml$ but FRG decreased LPS- and ConA-induced lymphocytes at $100\;{\mu}g/ml$. We showed that FRG increased the proliferation of HaCaT at 1, $10{\mu}g/ml$ but not by RG. These findings suggest that RG and FRG might have anti-inflammatory and anti-allergic effects, which can be needed to proper clinical concentration to applied to various allergic diseases and inflammation.

Inhibitory effect of Fagopyrum esculentum on degranulation and production of cytokine in RBL-2H3 cells (교맥의 RBL-2H3 비만세포 탈과립과 cytokine 생산 억제 효과)

  • Kang, Kyung-Hwa;Lee, Seung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
    • /
    • v.25 no.3
    • /
    • pp.1-12
    • /
    • 2012
  • Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

Differentially Up-expressed Genes Involved in Toluene Tolerance in Pseudomonas sp. BCNU106 (유기용매 내성 세균 Pseudomonas sp. BCNU106 균주에서 차별적으로 상향 발현되는 유전자군의 톨루엔 내성과의 연관성)

  • Joo, Woo Hong;Bae, Yun-Ui;Kim, Da Som;Kim, Dong Wan
    • Journal of Life Science
    • /
    • v.30 no.1
    • /
    • pp.88-95
    • /
    • 2020
  • Using a random arbitrarily primed polymerase chain reaction, messenger RNA expression levels were assessed after exposure to 10% (v/v) toluene for 8 hr in solvent-tolerant Pseudomonas sp. BCNU 106. Among the 100 up-expressed products, 50 complementary DNA fragments were confirmed to express repeatedly; these were cloned and then sequenced. Blast analysis revealed that toluene stimulated an adaptive increase in the gene expression level in association with transcriptions such as LysR family of transcriptional regulators and RNA polymerase factor sigma-32. The expression of catalase and Mn2+/Fe2+ transporter genes functionally associated with inorganic ion transport and metabolism increased, and the increased expression of type IV pilus assembly PilZ and multi-sensor signal transduction histidine kinase genes, functionally categorized into signal transduction and mechanisms, was also demonstrated under toluene stress. The gene expression level of beta-hexosaminidase in association with carbohydrate transport and metabolism increased, and those of DNA polymerase III subunit epsilon, DNA-3-methyladenine glycosylase II, DEAD/DEAH box helicase domain-containing protein, and ABC transporter also increased after exposure to toluene in DNA replication, recombination, and repair, and even in defense mechanism. In particular, the RNAs corresponding to the ABC transporter, Mn2+/Fe2+ transporter, and the β-hexosaminidase gene were confirmed to be markedly induced in the presence of 10% toluene. Thus, defense mechanism, cellular ion homeostasis, and biofilm formation were shown as essential for toluene tolerance in Pseudomonas sp. BCNU 106.

Purification and Characterization of β-N-Acetylhexosaminidase from Rice Seeds

  • Jin, Yu-Lan;Jo, Yu-Young;Kim, Kil-Yong;Shim, Jae-Han;Kim, Yong-Woong;Park, Ro-Dong
    • BMB Reports
    • /
    • v.35 no.3
    • /
    • pp.313-319
    • /
    • 2002
  • N-Acetyl-$\beta$-D-hexosaminidase ($\beta$-HexNAc'ase) (EC 3.2.1.52) was purified from rice seeds (Oryza sative L. var. Dongjin) using ammonium sulfate (80%) precipitation, Sephadex G-150, CM-Sephadex, and DEAE-Sephadex chromatography, sequentially. The activities were separated into 7 fractions($F_1-F_7$) by CM-Sephadex chromatography. Among them, F6 was further purified to homogeneity with a 13.0% yield and 123.3 purification-fold. The molecular mass was estimated to be about 52 kDa on SDS-PAGE and 37.4 kDa on Sephacryl S-300 gel filtration. The enzyme catalyzed the hydrolysis of both p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GlcNAc) and p-nitrophenyl-N-acetyl-$\beta$-D-hexosaminide (pNP-GalNAc) as substrates, which are typical properties of $\beta$-HexNAc'ase. The ratio of the pNP-GlcNAc'ase activity to the pNP-GalNAc'ase activity was 4.0. However, it could not hydrolyze chitin, chitosan, pNP-$\beta$-glucopyranoside, or pNP-$\beta$-glucopyranoside. The enzyme showed $K_m$, $V_{max}$ and $K_{cat}$ for pNP-GlcNAc of 1.65 mM, $79.49\;mM\;min^{-1}$, and $4.79{\times}10^6\;min^{-1}$, respectively. The comparison of kinetic values for pNP-GlcNAc and pNP-GalNAc revealed that the two enzyme activities are associated with a single binding site. The purified enzyme exhibited optimum pH and temperature for pNP-GlcNAc of 5.0 and $50^{\circ}C$, respectively. The enzyme activity for pNP-GlcNAc was stable at pH 5.0-5.5 and $20-40^{\circ}C$. The enzyme activity was completely inhibited at a concentration of 0.1 mM $HgCl_$ and $AgNO_3$, suggesting that the intact thiol group is essential for activity. Chloramine T completely inhibited the activity, indicating the possible involvement of methionines in the mechanism of the enzyme.

Effect of Scutellaria barbata Pharmacopuncture Extract on Degranulation and Inflammatory Mediator Release in RBL-2H3 Cells (반지련 약침 추출물의 RBL-2H3 세포 탈과립과 염증매개물질 분비 억제 효과)

  • Kwon, Hyuk-Sang;Song, Choon-Ho
    • Korean Journal of Acupuncture
    • /
    • v.29 no.3
    • /
    • pp.406-420
    • /
    • 2012
  • Objectives : Scutellaria barbata has been widely used in oriental medicine used for treatment of acute and chronic inflammatory diseases. In this study, to investigate the protective effect of Scutellaria barbata on type I allergic response, we determined whether Scutellaria barbata inhibits early or late allergic responses. Methods : To assess the effect of Scutellaria barbata Pharmacopuncture Extract(SB) in RBL-2H3 cells, we investigated the levels of the markers of degranulation such as ${\beta}$-hexosaminidase and histamine, inflammatory mediator such as IL-4, TNF-${\alpha}$, PGE2 and cysLT, and mRNA expression of cytokines and enzymes. In addition, we determined the levels of intracellular ROS by DCFH-DA assay and the free radical scavenging activity by DPPH method. Results : We found that SB suppressed the release of ${\beta}$-hexosaminidase and histamine and the production of IL-4, TNF-${\alpha}$, PGE2 and cysLT in RBL-2H3 by the antigen stimulation. SB also significantly inhibited the enzyme mRNA expressions, such as HDC2, COX-1, COX-2, 5-LOX and iNOS2, along with reduced cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, SB suppressed the levels of intracellular ROS. Conclusions : Our results indicate that SB protects against type I allergic response and exert an anti-inflammatory effect through the inhibition of degranulation, inflammatory mediator release and mRNA expression of cytokines and enzymes.

Screening and Evaluation of the Anti-allergic Effect of Korean Medicinal Plant Extracts (한국 약용식물자원 추출물의 항알레르기 활성 평가 및 선발)

  • Kim, Nan Sol;Choi, Doo Jin;Choi, Eun Ju;Lee, Jeong Hoon;Park, Saetbyul;Lee, Young Sup;Lee, Jae Won;Lee, Dae Young;Kim, Geum Sook;Lee, Seung Eun
    • Korean Journal of Medicinal Crop Science
    • /
    • v.26 no.1
    • /
    • pp.42-54
    • /
    • 2018
  • Background: Allergic diseases like such as atopic dermatitis, asthma, and rhinitis have recently increased both domestically and globally. The present study was undertaken to select candidates with anti-allergic activity from plant resources. Methods and Results: Fifty-six plant extracts at $20{\mu}g/m{\ell}$ were screened against ${\beta}$-hexosaminidase production and interleukin (IL)-4 release in degranulated rat basophilic leukemia (RBL)-2H3 cells. The anti-allergy activity of three plant extracts selected from the preliminary screening experiment, Polygonatum sibiricum F. Delaroche (root), Pyrus pytifolia var. culta (Makino) Nakai (leaf), and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud (root) were measured at concentrations of $2-250{\mu}g/m{\ell}$ in three cell lines as RBL-2H3, HaCaT and Jurkcat T cells. The assay showed the root extract of R. glutinosa to have an inhibitory activity of 4.2% - 28.6% on ${\beta}$-hexosaminidase production from IgE-sensitized RBL-2H3 cells. Each extract of P. sibiricum and R. glutinosa reduced IL-4 release in IgE-sensitized RBL-2H3 cells, respectively. The leaf extract of P. pyrifolia var. culta showed a significantly potent suppressive effect of 10.2% - 74.7% on the mRNA expression of tumor necrosis factor (TNF)-${\alpha}$ in HaCaT cells sensitized with TNF-a and INF-g, and showed inhibitory effect of -8.6% - 90.9% on the mRNA expression of IL-2 in Jurkat T cells sensitized with PMA and A23187. Conclusions: The results showed that the root of R. glutinosa and leaf of P. pyrifolia var. culta could be useful candidates as antiallergy materials.