• Title/Summary/Keyword: Hexosaminidase

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Inhibitory Effect of Persicaria perfoliata (L.) H. Gross on IgE Mediated Allergic Responses in RBL-2H3 Cells

  • Yoon, Hyun-Seo;Park, Chung-Mu
    • Journal of The Korean Society of Integrative Medicine
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    • v.8 no.4
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    • pp.163-169
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    • 2020
  • Purpose : This study aimed to investigate the anti-allergic effect of Persicaria perfoliata water extract (PPWE) on IgE stimulated rat basophilic leukemia (RBL-2H3) cell line. Methods : P. perfoliata (L.) H. Gross has been used in traditional medicine as an anti-allergic agent, antipyretic, and diuretic and for respiratory disorders. To analyze the anti-allergic activity of PPWE, release of β-hexosaminidase in RBL-2H3 cells was estimated by enzyme linked immunosorbant assay (ELISA). Also, the cytotoxic effect of PPWE was identified by WST assay, and nuclear factor (NF)-κB and its upstream signaling molecules were assessed by western blot analysis. Results : PPWE treatment significantly attenuated β-hexosaminidase release in a dose dependent manner without any cytotoxicity. PPWE inhibited β-hexosaminidase activity by 38.4±1.2, 36.6±0.6, 32.5±2.2 and 26.5±1.2 at 500, 250, 100, and 50 ㎍/㎖ of PPWE, respectively, compared with the control group. In addition, an analysis of the expression level of NF-κB, an inflammation transcription factor, in RBL-2H3 cells upon IgE stimulation provided reults consistent with the results of β-hexosaminidase release. The phosphorylated status of upstream signaling molecules for transcription factor, mitogen activated protein kinases (MAPKs), was also analyzed. The results showed that PPWE treatment dose-dependently inhibited phosphorylation of extracellular regulatory kinase (ERK) and c-Jun N-terminal kinase (JNK). These results show that PPWE had a strong IgE-mediated degranulation inhibitory effect on RBL-2H3 cells. Conclusion : P. perfoliata ameliorated IgE-mediated allergic reaction via the modulation of MAPK and NF-κB signaling pathway in RBL-2H3 cells. These results indicate that P. perfoliata could be a potential candidate for a treatment strategy against various allergic disorders.

Structure-activity Relationships of 4-Senecioyloxymethyl-6,7-dimethoxycoumarin Analogues as Anti-Allergic Agents

  • Jeong, Hye-Gwang;Lee, Jee-Hyun;Jung, Sang-Hun;Han, Eun-Hee;Kim, Joo-Hwan;Kim, Dong-Hee;Jin, Mi-Rim;Siripuram, Praveen Kumar;Choi, Yong-Seok;Song, Gyu-Yong
    • Bulletin of the Korean Chemical Society
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    • v.28 no.10
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    • pp.1725-1728
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    • 2007
  • Mast cells are key effector cells in the early phase allergic inflammation and in diverse immunological and pathological processes. In order to understand the effect on reduction of the anti-dinitrophenyl (DNP) IgE antibody-induced β-hexosaminidase release in RBL-2H3 rat mast cells, a novel series of 4-senecioyloxymethyl- 6,7-dimethoxycoumarins (SMDC) was prepared by reacting 4-chloromethyl-6,7-dimethoxycoumarin with various carboxylic acids. Compounds 8-11 with cyclic moiety such as phenyl, thiophenyl, pyridinyl, and furanyl group were found to inhibit-hexosaminidase release more potently (5.98-9.62 μM) than compounds 3- 7 and 12 with acyclic moiety (19.32-76.78 μM). Furthermore, compounds 8 and 9 inhibited IgE-induced ear swelling and significantly reduced systemic passive cutaneous anaphylaxis reaction in mice.

Experimental Study about Pathway of Aconiti Ciliare Tuber on Allergic Reaction of Inflammation (초오의 항알레르기 염증반응 및 기전탐색에 관한 연구)

  • Kim, Won-Ill
    • Korean Journal of Oriental Medicine
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    • v.16 no.3
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    • pp.155-166
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    • 2010
  • Objetives : The purpose of this study was to examine the pathway of anti-allergic effects of Aconiti Ciliare Tuber (ACT). Methods : We examined cell viability, ${\beta}$-hexosaminidase release, pro-inflammatory cytokines secretion and mRNA expressions, nuclear factor-kappa B (NF-${\kappa}B$) (p65) activation, inhibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) degradation, and MAPKs activation from RBL-2H3 cells pre-treatment by ACT of 1.0 mg/ml, 2.0 mg/ml separately. Results : We observed that ACT reduced the secretion of ${\beta}$-hexosaminidase, TNF-${\alpha}$, IL-4 and the expression of COX-2 mRNA in RBL-2H3 cells. Futhermore, ACT inhibited the levels of activation of NF-${\kappa}B$ (p65) protein, ERK MAPK, and degradation of $I{\kappa}B-{\alpha}$ in RBL-2H3 cells. Conclusions : These results show that ACT has an anti-histamine effect and inhibitory effect of NF-${\kappa}B$ (p65) through regulation of $I{\kappa}B-{\alpha}$ degradation. This improves that ACT could be used as an anti-allergic medicine.

The Antiallergic and Antioxidative Effects of Chaenomeles Sinensis (CS) in RBL 2H3 Cells (목과(木瓜) 추출물이 항알레르기 및 항산화 작용에 미치는 영향)

  • Lee, Seung-Yeon
    • The Journal of Pediatrics of Korean Medicine
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    • v.24 no.2
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    • pp.126-136
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    • 2010
  • Objectives The purpose of this study was to investigate the antiallergic and antioxidative effects of Chaenomeles Sinensis (CS). Methods CS pretreatments inhibited anti-DNP IgE in RBL-2H3 mast cells for an hour. we measured cell viability, $\beta$-hexosaminidase release, IL-4, TNF-$\alpha$ secretion, and IL-4, TNF-$\alpha$ mRNA expression CS pretreatments inhibited DNP-HSA($10\;{\mu}g/ml$) for ten minutes, we measured Dicholrodihydrofluorescein(DCF) and DPPH radical-scavenging activity in 0.5 mM 1,1-diphenyl-2-picrydrazyl(DPPH) radical solution, 0.1ml, 99% ethanol 0.8ml, and CS 0.1 ml mixed solution. Results 0, 1, 2, 3, 4, 5 mg/ml CS treatments were not affect on cell viability and inhibited b-hexosaminidase release, IL-4, TNF-$\alpha$ secretion, CS treatments also decreased IL-4, TNF-$\alpha$ mRNA expression in RBL-2H3 cells. CS treatments inhibited reactive oxygen species(ROS) and DPPH radical-scavenging activity. Conclusions These results suggest that CS may be useful for the prevention or treatment of allergic disease.

Experimental Study about the Pathway of Inflammatory Allergic Reaction of Cheonmaec-tang (천맥탕(天麥湯)의 알레르기 염증반응 및 기전탐색에 관한 연구)

  • Kim, Yong-Hyeon;Lee, Seung-Yeon;Kim, Won-Il
    • The Journal of Pediatrics of Korean Medicine
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    • v.24 no.1
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    • pp.93-103
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    • 2010
  • Objectives The purpose of this study was to examine the pathway of anti-allergic effects of Cheonmaec-tang (CMT). Methods We examined the cell viability, $\beta$-hexosaminidase release, pro-inflammatory cytokines secretion and mRNA expressions, nuclear factor-kappa B (NF-${\kappa}B$) (p65) activation, inbibitor kappa B-alpha ($I{\kappa}B-{\alpha}$) degradation, and MAPKs activation in RBL-2H3 cells pre-treated by CMT of 2.0 mg/ml, 4.0 mg/ml separately. Results We observed that CMT reduced the secretion of $\beta$-hexosaminidase, TNF-$\alpha$, IL-4 and the expression of COX-2 mRNA in RBL-2H3 cells. Furthermore, CMT inhibited the levels of activation of NF-${\kappa}B$ (p65) protein, ERK MAPK, and degradation of $I{\kappa}B-{\alpha}$ in RBL-2H3 cells. Conclusions These results show that CMT has an anti-histamine effect and inhibitory effect of NF-${\kappa}B$ (p65) through regulation of $I{\kappa}B-{\alpha}$ degradation. These suggest that CMT could be used as an anti-allergic medicine.

Inhibitory Effect of Alpiniae officinarum Rhizoma Extract on Degranulation in RBL-2H3 Cells

  • Kim, Eunhee;Ahn, Sejin;Lee, Deug-Chan
    • Korean Journal of Plant Resources
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    • v.28 no.3
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    • pp.321-328
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    • 2015
  • Alpiniae officinarum Rhizoma (the rhizome of Alpinia officinarum Hance, known as lesser galangal), a family of Zingiberaceae, has been used to reduce pain of infection and inflammatory diseases in Asian countries. The present study was focused to evaluate the inhibitory degranulation effect of Alpiniae officinarum Rhizoma extract in RBL-2H3 rat basophilic leukemia cells. Cell viability was measured by MTT assay. RBL-2H3 cells were stimulated by phorbol 12-myristate 13-acetate and calcium ionophore A23187. Mast cell degranulation was analyzed by measuring release of β-hexosaminidase in RBL-2H3 cell. Gene expression was measured by qRT-PCR and signaling molecules were detected by immunoblotting. The Alpiniae officinarum Rhizoma extract suppressed β-hexosaminidase release in dose-dependent manner and inhibited cycloxygenase-2 and tumor necrosis factor-α gene expression. Furthermore, it was found that Alpiniae officinarum Rhizoma extract reduced mitogen-activated protein kinases, especially phosphorylated p38, at 0.75 ㎎/㎖ of Alpiniae officinarum Rhizoma extract concentrations. These data show that Alpiniae officinarum Rhizoma extract has immunosuppressive effect in mast cell induced allergic inflammation.

Anti-allergic Actions of the Leaves of Castnea crenata and Isolation of an Active Component Responsible for the Inhibition of Mast Cell Degranulation

  • Lee, Eun;Choi, Eun-Ju;Cheong, Ho;Kim, Young-Ran;Ryu, Shi-Yong;Kim, Kyeong-Man
    • Archives of Pharmacal Research
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    • v.22 no.3
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    • pp.320-323
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    • 1999
  • The anti-allergic actions of the leaves of Castanea crenata (Fagaceae) were studied. The water extract demonstrated potent anti-allergic actions in in vivo and in vitro experiments. The oral or intraperitoneal administration of the extract (100 or 200 mg/kg) caused a significant inhibition of the 48 hr-PCA (up to 90%) and the vascular permeability induced by histamine or serotonin in rats (about 80%). The anaphylactic release of ${\beta}$-hexosaminidase for RBL-2H3 cells was also significantly inhibited by the extract in as dose-dependent manner with an IC50 value of 230 $\mu\textrm{g}$/ml. The activity-guided fractionation of the extract, based on the determination of inhibitor effect upon the release of ${\beta}$-hexosaminidase, led to the isolation of quercetin as an active principle responsible for the inhibition of degranulation.

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Inhibitory Effect of Rhododendron Mucronulatum Root Extract on Allergic Inflammation (진달래 뿌리 추출물의 알레르기 염증 억제 효과)

  • Jang, Si Sung;Lee, DaeJoong;Song, Jihoon;Park, Do Hwi;Jeon, Chan Yong;Hwang, Gwi Seo
    • The Journal of Internal Korean Medicine
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    • v.43 no.1
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    • pp.68-78
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    • 2022
  • Objective: In this study, we investigated the protective effect of rhododendron mucronulatum extract (RME) on allergic reactions and inflammation. Methods: The effect of RME was determined using ELISA and RT-PCR in RBL-2H3 mast cells and RAW 264.7 macrophage cells. We determined cell viability, β-hexosaminidase release, and the synthesis of IL-4 and TNF-α in RBL-2H3 cells. In addition, we determined NO from RAW 264.7 and the gene expression of IL-1β, iNOS, IL-6, TNF-α, and IL-10. Results: RME inhibited β-hexosaminidase release and synthesis of IL-4 and TNF-α in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. In addition, RME inhibited the production of NO and the gene expression of IL-1β, iNOS, IL-6, and TNF-α in LPS-stimulated RAW 264.7 cells. Conclusion: From these results, we concluded that RME possesses anti-allergic activity and anti-inflammatory activity due to the inhibition of mast cells and macrophage function.

Inhibition of Interleukin-4 and β-Hexosaminidase Release in RBL-2H3 Cells by Compounds Isolated from Lobelia chinensis

  • Kim, Tae Young;Jo, Beom-Geun;Park, No-Jun;Park, Young-Hun;Kim, Su-Nam;Yang, Min Hye
    • Natural Product Sciences
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    • v.27 no.4
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    • pp.251-256
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    • 2021
  • Lobelia chinensis Lour. has commonly been used in traditional Chinese medicine for the treatment of antidote, diuretic, diarrhea, and inflammation. This study aimed to identify the active compounds in an aqueous extract of L. chinensis responsible for its anti-atopic effect in vitro using RBL-2H3 cells. A chemical investigation of secondary metabolites in an aqueous extract of L. chinensis led to the isolation of nine chemical constituents, which included the four marker compounds, and these were evaluated for their inhibitory effects on IL-4 mRNA expression and the release of β-hexosaminidase in propidium iodide-induced RBL-2H3 cells. We found diosmetin and fraxidin inhibited cellular IL-4 mRNA expression, and that diosmetin and 6,8-dimethoxycoumarin inhibited DNP-specific IgE-induced degranulation in these cells. Our study suggests that diosmetin, fraxidin, and 6,8-dimethoxycoumarin are potential candidates for the treatment of atopic diseases.

Pharmacological potential of Cordyceps militaris with enhanced Cordycepin production for anti-inflammatory and tumor cell anti-proliferative applications

  • Ha, Si-Young;Jung, Ji-Young;Yang, Jae-Kyung
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.93-101
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    • 2022
  • Cordyceps militaris mycelium extracts containing high amounts of cordycepin were evaluated in vitro for their anti-inflammatory and tumor cell growth-inhibitory activities. All extracts dose dependently inhibited the increased production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), and 𝛽-hexosaminidase in lipopolysaccharide (LPS)-stimulated inflammatory cells. All extracts were evaluated for anti-proliferative activity against normal RBL-2H3 cells and diverse types of cancer cell lines, including HCT, MC5-7, U-87MG, AGS, and A549 cells. The extract showed the strongest growth inhibition (IC50 = 28.13 ㎍/mL) relative to vehicle-treated control cells against fibrosarcoma (MC5-7). We have demonstrated anti-inflammatory activity of C. militaris via inhibition of NO, ROS production, and 𝛽-hexosaminidase release in activated cells. C. militaris mycelium extract was also evaluated mechanistically and found to exert six types of anti-cancer activity, confirming its pharmacological potential. Our study suggests C. militaris use as a potential source of anti-inflammatory and anti-cancer agents. C. militaris may also be considered a functional food.