• Title/Summary/Keyword: RBL 2H3 cell

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Anti-allergic Effect of Bopyeoyangyeong-jun (보폐양영전(保肺養營煎)이 알레르기 염증반응에 미치는 영향)

  • Kim, Hong-Gi;Lee, Jae-Hyuk;Park, Sung-Jun;Kim, Jin-Young;Kim, Bum-Hoi;Park, Dong-Il
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.3
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    • pp.660-665
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    • 2008
  • In the present study, we investigated the anti-allergic effect of the water extract of Bopyeoyangyeong-jun(BYJ). To investigate the biological effect of BYJ, We examined cytotoxicity and inflammatory cytokine secretion with RBL-2H3. We examined Cell Viability, ${\beta}$-hexosaminidase, tumor necrosis factor-alpha(TNF-${\alpha}$), interleukin(IL)-4 secretion from RBL-2H3 cell after pre- treatment with Bopyeoyangyeong-jun of 1 mg/ml, 2 mg/ml. RBL-2H3 cell was stimulated with phorbol 12-myristate 13-acetate(PMA) and calcium ionophore A23187. BYJ by itself had no effect on viability of RBL-2H3 cell. We observed that Bopyeoyangyeong-jun reduced ${\beta}$-hexosaminidase, TNF-${\alpha}$, IL-4 secretion in RBL-2H3 cell. Taken together, these results indicate that Bopyeoyangyeong-jun has anti-histamic effect and controls TNF-${\alpha}$, IL-4 secretion on allergic reaction.

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 Artemisiae Asiaticae Herba on Degranulation, Production of Cytokine, and $Fc{\varepsilon}RI$ Expression in RBL-2H3 Cells (애엽(艾葉)의 RBL-2H3 비만세포 탈과립, cytokine 생산 및 $Fc{\varepsilon}RI$ 유전자 발현에 미치는 영향)

  • Ban, Hak Jin;Park, Dong II;Kang, Kyung Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.6
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    • pp.915-921
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    • 2012
  • Artemisiae Asiaticae Herba (AAH) has been used to remedy of symptoms such as bleeding, dysmenorrhea, eczema and itchy skin in Oriental Medicine. In this study, we investigated the protective effect of AAH on allergic response. The effect of AAH was analyzed by ELISA and RT-PCR in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase and histamine as markers of degranulation, production of IL-4 and TNF-${\alpha}$, and gene expression of HDC2, cytokines and $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit. We found that AAH suppressed ${\beta}$-hexosaminidase and histamine release, the production of IL-4 and TNF-${\alpha}$ in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. AAH also significantly decreased cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12, IL-13, TNF-${\alpha}$, and GM-CSF, and increased cytokine mRNA expressions of IL-10 in RBL-2H3. In addition, AAH suppressed mRNA expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit on cell surface. Our results indicate that AAH protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit.

Anti-Allergic Effects of Angelica gigas Nakai and Corni fructus extract (AC) on degranulation and production of cytokine in RBL-2H3 mast Cells (RBL-2H3 세포에서 당귀(當歸) 및 산수유(山茱萸) 복합추출물의 알레르기 개선에 대한 효과)

  • Tae Woo Oh
    • Herbal Formula Science
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    • v.31 no.4
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    • pp.315-325
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    • 2023
  • Objectives : Recently, research has been actively conducted on the efficacy of complexes based on oriental medicine prescriptions for improving immune activity and allergies. In this study, In this study, we aimed to examine the effect of Angelica gigas Nakai and Corni fructus extract (AC), medicinal herbs, among candidate drugs derived through preliminary experiments with various components of oriental medicine prescriptions for allergies, on allergies in RBL-2H3 cells. Methods : We evaluated the effect of the ethanol extract of Ulmus on the allergic inflammatory response in anti-DNP-IgE activated DNP-HSA in RBL-2H3 cells. Cell toxicity was determined by WST-1 assay and the markers of degranulation such as beta-hexosaminidase, histamine, TNF-α and IL-6 production of inflammatory mediators and FcεRI-mediated expression. Results : The results showed that treatment with AC extract (20, 40 and 80㎍/㎖) noncytotoxic levels and significantly inhibited the release of β-hexosaminidase, histamine and the production of TNF-α and IL-6 in RBL-2H3 by the antigen stimulation. Conclusions : These results indicate that AC extract exhibits anti-allergic activity through inhibition of degranulation and inhibition of inflammatory mediators and cytokine release. These findings suggest that AC extract may have potential as a prophylactic and therapeutic agent for the treatment of various allergic diseases.

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.

Identification of Functionally Different Rat IgE in RBL-2H3 Exocytosis

  • Kim, Jin-Sub;Cho, Sungae;Joo, Kyoung-Hwan;Lee, Joon-Sang;Conrad, Daniel H.;Cho, Sung-Weon
    • IMMUNE NETWORK
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    • v.2 no.4
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    • pp.195-201
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    • 2002
  • Background: IgE is closely related to the development of allergies. However, the poor relationship between the specific IgE level and the severity of allergic diseases suggests the possibility of functionally different IgE isoforms. With this in mind, rat basophilic leukemia (RBL)-2H3 activation was analyzed with each type of rat IgE for two parameters, exocytosis and IL-4 mRNA production. RBL-2H3 has been well documented in the rat mucosal mast cell line. Methods: RBL-2H3 cells sensitized with each kind of rat IgE was activated by cross-linking FcRI with B5 (monoclonal anti-rat IgE mouse IgG antibodies). The RBL-2H3 exocytosis was measured by analyzing the ${\beta}$-hexosaminidase level, and the level of IL-4 mRNA synthesis was analyzed using semiquantitative RT-PCR. Rat IgE, which was produced by a parasite infection (REP), was prepared using either Paragonimus westermani metacercariae (REP-PW) or Anisakis simplex third stage larvae (REP-AS). A rat IgE prototype of IR162 was prepared by a peritoneal injection of immunocytoma. Results: The level of exocytosis showed a linear relationship with the rat IgE concentration when REP-PW or REP-AS was applied. However, it exhibited a biphasic response with IR162. In addition, the time course of heating at $56^{\circ}C$ illustrated the similarity between REP-PW and REP-AS, which differed from that of IR162. In contrast, the level of IL-4 mRNA synthesis in the RBL-2H3 cells with IR162 was comparable to that of either REP-PW or REP-AS. Conclusion: These results suggest that functionally different rat IgE isoforms exists in RBL-2H3 exocytosis.

The Anti -allergic Effects of Taraxaci Herba(蒲公英) on the RBL-2H3 Cells (RBL-2H3 세포에서 포공영의 항알레르기 효과)

  • Jo, Ju-Heum;Lyu, Ji-Hyo;Kim, Cheol-Hong;Kang, Kyung-Hwa;Yoon, Hwa-Jung;Lee, Seung-Yeon;Ko, Woo-Shin;Kim, Won-Il
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.20 no.1 s.32
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    • pp.209-217
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    • 2007
  • 청열해독(淸熱解毒) 작용을 하는 포공영의 항알러지 효과를 알아보기 위하여 RBL-2H3 cell line에 PMA와 A23187을 자극한 후 RBL-2H3 cell내 과립의 degranulation의 억제정도를 살펴보기 위하여 ${\beta}-Hexosaminidase$의 누출량의 억제 정도를 살펴본 결과 포공영은 농도 의존적으로 현저하게 degranulation을 억제하였다. 알러지 반응 시 세포내에서 분비되어 중요한 작용을 하는 것으로 알려진 사이토카인인 $TNF-{\alpha}$, IL-4의 합성조절 정+도를 살펴본 결과, $TNF-{\alpha}$, IL-4의 mRNA 수준에서의 발현의 조절을 통한 합성도 억제하였다. 이 결과로 보아 포공영은 알레르기와 관련된 질환에 치료약물로서 응용이 가능할 것으로 사료된다.

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Identification of Calcium/Calmodulin-Dependent Phosphatase as the Dephosphorylating Enzyme of IgE-Dependent Histamine-Releasing Factor in RBL-2H3 (RBL-2H3 세포에서 IgE-depnedent Histamine-releasing Factor의 탈인산화 효소에 관한 연구)

  • Hwang Sun-Ok;Lee Kyunglim
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.189-193
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    • 2005
  • IgE-dependent histamine-releasing factor(HRF) was initially described as a secretagogue for secretion of histamine from IgE+ basophils from a subset of allergic donors. Previously, we identified that S98 residue of HRF was phosphorylated using anti-HRFpS98 antibody which specifically recognizes the phosphorylated serine residue of HRF and HRFS98A mutant construct. In vitro kinase assay, only wild type HRF was phosphorylated by PKC, and S98A HRF was not affected by PKC. In this study, we attempted to characterize the phosphatase which specifically dephosphorylates HRF by immunoprecipitation and pull-down assay. In RBL-2H3 cells, HRF interacted only with calcineurin (also called as PP2B, calcium/calmodulin-dependent phosphatase) but not with PP1 or PP2A. The results suggest that HRF is most likely dephosphory-lated by calcineurin.

Fagopyrum esculentum Extract Suppresses the Release of Inflammatory Mediator and Proximal Signal Events in $Fc{\varepsilon}RI$-mediated RBL-2H3 Cell Activation (교맥(蕎麥)의 비만세포 염증매개물질의 분비와 $Fc{\varepsilon}RI$ 신호전달에 미치는 효과)

  • Kang, Kyung-Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.4
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    • pp.469-474
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    • 2012
  • Fagopyrum esculentum(FE) is an important food crop and medicinal plant that is used to improve diabetes, obesity, hypertension, hypercholesterolemia and constipation in Korea, but the underlying mechanisms involved in its anti-allergic activity are not fully understood. We investigated the effects on the release of inflammatory mediator and proximal signal events in $Fc{\varepsilon}RI$-mediated RBL-2H3 cell activation. FE reduced antigen (DNP-HSA)-induced release of histamine, prostaglandin D2 (PGD2) and cysteinyl Leukotriene (cysLT) in IgE-sensitized RBL-2H3 cells. In addition, it inhibited antigen-induced HDC2 and COX-2 and 5-LO mRNA expression in IgE-sensitized RBL-2H3 cells. FE also suppressed antigen-induced $Fc{\varepsilon}RI{\beta}$ and $Fc{\varepsilon}RI{\gamma}$ subunit mRNA expression in these cells. To identify the mechanisms underpinning the inhibition of release of inflammatory mediators such as histamine and PGD2 and cysLT by FE, we examined the proximal signal events of intracellular FceRI signaling molecules. FE suppressed antigen-induced phosphorylation of Lyn, Syk, LAT, $PLC{\gamma}1$, PI3K, Akt and cPLA2. Collectively, the anti-allergic effects of FE in vitro suggest its possible therapeutic application to inflammatory allergic diseases, in which its inhibition of inflammatory mediator and FceRI-dependent signaling events in mast cells may be hugely beneficial.

Panax ginseng-derived fraction BIOGF1K reduces atopic dermatitis responses via suppression of mitogen-activated protein kinase signaling pathway

  • Lorz, Laura Rojas;Kim, Donghyun;Kim, Mi-Yeon;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.44 no.3
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    • pp.453-460
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    • 2020
  • Background: BIOGF1K, a fraction of Panax ginseng, has desirable antimelanogenic, anti-inflammatory, and antiphotoaging properties that could be useful for treating skin conditions. Because its potential positive effects on allergic reactions in skin have not yet been described in detail, this study's main objective was to determine its efficacy in the treatment of atopic dermatitis (AD). Methods: High-performance liquid chromatography was used to verify the compounds in BIOGF1K, and we used the (3-4-5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide method to determine its cytotoxicity in RBL-2H3 and HMC-1 cell lines. RBL-2H3 cells were induced using both anti-DNP-IgE/DNP-BSA and calcium ionophore (A2187) treatments, whereas HMC-1 cells were induced using A2187 alone. To measure mast cell degranulation, we performed histamine (enzyme-linked immunosorbent assay) and β-hexosaminidase assays. To quantify interleukin (IL)-4, IL-5, and IL-13 levels in RBL-2H3 cells, we performed quantitative polymerase chain reaction (PCR); to quantify expression levels of IL-4 and IL-13 in HMC-1 cells, we used semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Finally, we detected the total and phosphorylated forms of extracellular signal-regulated kinase, p-38, and c-Jun N-terminal kinase proteins by immunoblotting. Results: BIOGF1K decreased the AD response by reducing both histamine and β-hexosaminidase release as well as reducing the secretion levels of IL-4, IL-5, and IL-13 in RBL-2H3 cells and IL-4 and IL-13 in HMC-1 cells. In addition, BIOGF1K decreased MAPK pathway activation in RBL-2H3 and HMC-1 cells. Conclusions: BIOGF1K attenuated the AD response, hence supporting its use as a promising and natural approach for treating AD.