• Title/Summary/Keyword: RBL-2H3 cells

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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
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    • v.29 no.3
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    • pp.406-420
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    • 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.

Effects of Protein Kinase Inhibitors on Histamine Release and ROS Generation in RBL 2H3 Cells

  • Yoon, Mi-Yun;Cho, Nam-Young;Lee, Ji-Yun;Seo, Moo-Hyun;Kim, Chang-Jong;Sim, Sang-Soo
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.297.2-297.2
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    • 2002
  • Previous report showed that histamine release by HCI was mediated via reactive oxygen species (ROS) generation in RBL 2H3 cells. To investigate action of protein kinase on histamine release and ROS generation. we observed effects of protein kinase inhibitors on histamine release and ROS generation in RBL 2H3 cells stimulated by HCI HCI dose-dependently increased both histamine release and ROS generation. HCI-induced histamine release was significantly inhibited by bisindolmaleimide (10 ${\mu}$M). DHC (10 ${\mu}$M). , and wortmannin (10 ${\mu}$M), but not by PD098059 (10 ${\mu}$M). ON the other hand. HCI-induced ROS generation was significantly inhibited by DHC (10 ${\mu}$M). but not by bisindolmaleimide(10 ${\mu}$M). wortmannin (10 ${\mu}$M). and PD098059 (10 ${\mu}$M). However KN-62 did not inhibited both. These results showed that involvement of protein kinase in regulation of histamine release and ROS generation may be different and only tyrosine kinase may be associated with regulation of both histamine release and ROS generation in RBL 2H3 cells.

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Suppressive Effect of CheongGiGeoYangTang on Allergic Inflammation of RBL-2H3 Mast Cells and OVA/alum-sensitized Mice (청기거양탕(淸肌祛痒湯)의 RBL-2H3 비만세포와 OVA/alum 감작 생쥐의 알레르기 염증 반응 억제 효과)

  • Lee, Hye-Lim;Han, Jae-Kyung;Kim, Yun-Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.26 no.1
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    • pp.1-15
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    • 2012
  • $CheongGiGeoYangTang$ has been used for anti-allergenic purpose. However there was no experimental study about its effect. Therefore, this study was designed to investigate the anti-allergenic effect of $CheongGiGeoYangTang$. Methods Modifiability of RBL-2H3 mast cells' IL-4, IL-13 was analyzed by qRT-PCR and ELISA. Also, the suppressive effect of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}B$ p65 transcription factors was observed by western blotting. OVA-specific IgE, IL-4 and IL-13 production in ovalbumin allergy model was examined as well. Results It was showed that the RBL-2H3 mast cells treated with $CheongGiGeoYangTang$ extract(CGGYT) was significantly suppressed mRNA expression, production of IL-4 and IL-13, and prominently inhibited the expression of transcription factors including GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-${\kappa}B$ p65 but not c-Jun. The administration of CGGYT was suppressed the amount of OVA-specific IgE, IL-4 and IL-13 in OVA/alum-sensitized mice. Conclusions We considered CGGYT would regulate the allergic inflammation as inhibition of IL-4 and IL-13 production in activated mast cells and Th2 cells.

Induction of Cytotoxicity by HexamethyleneBisacetamide is Associated with the Activity of Telomerase in RBL-2H3 Cells (RBL-2H3 세포에서 hexamethylene bisacetamide의 세포독성과 telomerase 활성과의 관계)

  • Kim Young Mi
    • Environmental Analysis Health and Toxicology
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    • v.19 no.3
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    • pp.235-240
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    • 2004
  • 본 연구는 비만세포의 일종인 RBL-2H3 세포주에 대한 hexamethylene bisacetamide (HMBA)의 세포독성 유도 기전이 telomerase의 활성과 관계가 있다는 것에 대한 보고이다. RBL-2H3 세포주에 5mM HMBA를 처리시 그 세포주의 세포성장이 억제 되었다. 기전연구로서 HMBA는 telomerase활성을 억제 하였으며 RT-PCR에 의한 결과 telomerase mRNA발현의 억제에 의한 것이었다. Madl은 telomerase의 발현을 억제하는 억제인자로 잘 알려져 왔으며 이를 확인해본 결과 Madl은 발현은 유의성있게 증가 되었다. 이를 종합해보면, HMBA에 의한 RBL-2H3 세포주에 대한 세포독성은 Madl 발현의 증가에 따른 telomerase발현의 억제에 기인하는 것으로 밝혀졌다.

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.

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.

Effects of Tribuli Fructus extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells (비만세포에서 백질려 추출물의 항염증효과에 대한 연구)

  • Rho, Hyo Sun;Park, Yong-Ki;Bae, Hyo Sang
    • The Korea Journal of Herbology
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    • v.32 no.2
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    • pp.107-114
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    • 2017
  • Objectives : Tribulus terrestris $Linn{\acute{e}}$ (Tribuli Fructus; TF) has been used to treat hypochondrium, agalactia, nebula, itching and vitiligo in traditional Korean medicine. In this study, we investigated the effects of TF 30% ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : TF extract was prepared by 30% ethanol. RBL-2H3 cells, a rat mast cell line, were treated with TF extract at different concentrations for 1 hr and then stimulated with DNP-IgE/HSA for indicated times. Cell viability was measured by WST-1 assay. The expression of inflammatory cytokines (IL-4, IL-13 and $IFN-{\gamma}$) mRNA was determined by reverse transcriptase-PCR, and the phosphorylation of ERK1/2, p38 and JNK MAP kinases (MAPKs) was determined by Western blot. The nuclear expression of $NF-{\kappa}B$ p65 in the cells was detected by Western blot and immunocytochemistry, respectively. Results : The treatment of TF extract at 0.1 and $0.2mg/m{\ell}$ significantly decreased the expression of IL-4 and IL-13 mRNA in IgE-stimulated RBL-2H3 mast cells, while significantly increased the expression of $IFN-{\gamma}$ mRNA. TF extract treatment was also inhibited the phosphorylation of ERK1/2, p38 and JNK MAPKs in IgE-stimulated RBL-2H3 mast cells in a dose-dependent manner. In addition, TF extract significantly blocked the translocation of $NF-{\kappa}B$ p65 into the nuclear of cells after IgE stimulation. Conclusions : These results indicate that TF extract inhibits inflammatory response in IgE-stimulated mast cells through blocking MAPKs/$NF-{\kappa}B$ pathway. This suggests that TF extract has an anti-inflammatory activity in mast cell activation.

The Study of Seunggal-tang Gamibang on the Anti-allergy Effects

  • Ju, Bong-Hyun;Lyu, Sun-Ae;Ko, Woo-Shin;Kim, Own-Il;Lee, Seung-Yeon
    • The Journal of Pediatrics of Korean Medicine
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    • v.21 no.2
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    • pp.109-119
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    • 2007
  • Objectives The purpose of this study is to investigate effects of Seunggal-tang gamibang (SGT) on the PMA plus A23187-induced RBL-2H3 cells. Methods The effect of SGT on ${\beta}$ -hexosamonidase release, passive cutaneous anaphylaxis, $TNF-{\alpha}$ production and IL-4 secretion was studied. Results SGT inhibited the release of ${\beta}$ -hexosamonidase in RBL-2H3, without affecting cell viability. The passive cutaneous anaphylaxis (PCA) was inhibited by the SGT. Moreover SGT also reduced production of $TNF-{\alpha}$ and secretion of IL-4 in a dose-dependent manner. Conclusions SGT has anti-allergy effects on the RBL-2H3 cells.

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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.

Suppression of Th2-associated Cytokines Expression by Schizandrin in Basophils and Mast Cells (Schizandrin에 의한 호염구와 비만세포의 Th2 관련 사이토카인 발현 억제)

  • Jeong, Hwa-Hyun;Yoon, Soo-Jeong;Pyo, Myoung-Yun
    • Korean Journal of Pharmacognosy
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    • v.43 no.3
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    • pp.224-230
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    • 2012
  • Schizandrin (SZ), was isolated from the fruit of Schizandra chinensis, has been reported to have many biological properties, including anti-inflammatory and antitumor activities. However, its anti-allergic effects are not completely elucidated. We focused on the anti-allergic effects of SZ in PMA/ionomycin (PI)-induced rat basophilic leukemia (RBL-2H3) cells and P815 mast cells. Cytokines (IL-4, IL-13), synthesized by basophils and mast cells, are implicated in pathological conditions such as asthma and allergy. The production of IL-4 and IL-13 was quantified by ELISA and the mRNA expression was detected by using RT-PCR assay. In this study, we found that SZ did not show cytotoxic effect at up to 100 ${\mu}M$ on RBL-2H3 cells and mast cells. In addition, SZ inhibited the production of IL-4 and IL-13 and also decreased the level of mRNA in PI-induced RBL-2H3 cells and mast cells in a dose-dependent manner. Thus, we suggest that SZ may have the effect on preventing allergic disorders by inhibiting IL-4 and IL-13 cytokines.