• Title/Summary/Keyword: DNP-IgE

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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.

The Effects of Dictamni Radicis Cortex Extracts on DNCB Induced Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse (백선피(白鮮皮) 추출물이 DNCB로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향)

  • Lee, Youn Shil;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.28 no.4
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    • pp.1-29
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    • 2014
  • Objectives Dictamni Radicis Cortex extracts (DRC) has been known to suppress allergic reaction, however the cellular target of DRC and its mode of action remain unclear. The purpose of this study is to investigate the effects of Dictamni Radicis Cortex extracts on DNCB induced atopic dermatitis-like skin lesions of NC/Nga mouse. Methods This study was designed to investigate the effects of DRC extract in the DNP-IgE-induced activation of MC/9 murine mast cell lines in vitro and in the DNCB-induced activation of NC/Nga mouse in vivo. For this investigation, We examined IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA analysis and manifestations of NFAT1, NFAT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting in vitro. Then, we examined WBC, eosinophil and neutrophil in NC/Nga mouse, IL-5, IL-13 in serum, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $^+Gr-1^+CD11b$, $B220^+CD23^+$ in the ALN, PBMCs and dorsal skin, IL-5, IL-13 in the dorsal skin by Real-time PCR and the distribution of mast cells by H&E and toluidine blue. Results In vitro the mRNA expression of IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$, GM-CSF and IL-13, MIP-$1{\alpha}$ production by ELISA analysis were completely abolished by DRC and the western blot analysis decreased the expression of mast cell-specific transcription factors including NFAT-1, NF-${\kappa}B$ p65. In vivo DRC oral adminstration also decreased the counts of WBC, eosinophils and inflammatory cytokines such as IL-13 and IgE in the serum. DRC oral adminstration elevated IL-4 level in the spleenocyte culture supernatant. DRC oral adminstration decreased total ALN cells, total skin cells, cell numbers of $CD4^+$, $B220^+CD23^+$ in the ALN, $^+Gr-1^+CD11b$ in the PBMCs and $CD4^+$, $CD8^+$ in the dorsal skin. The mRNA expression of IL-5, IL-13, thickness of epidermis, inflammation immune cells and mast cells were abolished by DRC in the dorsal skin. Conclusions Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mouse were much improved by DRC oral adminstration. These results, therefore, suggest that DRC can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis induced in NC/Nga mouse, and may play an important role in recovering AD symptoms.

Effects of Ethanol Extract of Liriope platyphylla on Allergic Inflammation (맥문동(麥門冬) 에탄올 추출물이 알레르기 염증 반응에 미치는 영향)

  • Kim, Ho Seok;Park, Ji Hye;Kim, Hyun Kab;Kim, Jae Hyun;Lee, Bina;Min, Ju Hee;Kim, Eun Young;Jung, Hyuk Sang;Lee, Hyang Sook;Sohn, Young Joo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.5
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    • pp.512-519
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    • 2014
  • This study investegated the effect of Liriope platyphylla (LP) on allergic reactions and its mechanism of action. We investigated the effect of LP on Evans Blue (EB) extravasation induced by anti-dinitrophenyl (DNP)-IgE in rats. We tested whether the ethanol extract of LP reduced ear skin thickness and historical changes induced by topical application of 2,4-dinitrofluorobenzene (DNFB) to ears of mice. We evaluated compound 48/80-induced release of histamine in rats peritoneal mast cell (RPMCs). We also investigated the regulatory effect of LP on the level of inflammatory mediators in PMACI-induced human mast cell (HMC-1); cytokine IL-6, IL-8, TNF-${\alpha}$ in HMC-1, MAPKs (ERK, JNK and p38) in HMC-1. The ethanol extract of LP (81.3 mg/100 g body weight) significantly inhibited the PCA reaction compared with the control (P < 0.05). However, LP did not prevent topical applications of DNFB-induced ear skin thickening and histological changes. In RPMCs, histamine release induced by compound 48/80 was significantly attenuated by LP at $100{\mu}g/ml$ (P < 0.05). LP extract ($100{\mu}g/ml$) significantly reduced the PMACI-induced IL-6, IL-8, and TNF-${\alpha}$ secretion via inhibition of ERK phosphorylation in HMC-1. In conclusion, the ethanol extract of LP inhibited mast cell-derived, immediate-type allergic reactions, and the result suggest the potential of LP for preventing allergic inflammatory disorders.

Inhibitory Effects of Immediate-Type Allergic Reaction of Okbyungpoongsan-Gami by Anal Therapy (항장효법에 의한 옥병풍산가미의 즉각형 알레르기 반응 억제 효과)

  • Cho Jeong Yeon;Moon Gu;Moon Suk Jae;Won Jin Hee;You Kyoung Tae;Lee Jong Deok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.239-244
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    • 2002
  • Okbyungpoongsan-Gami (OG) has been used for the treatment of excessive sweating with general weakness and allergic rhinitis recently. Anal therapy is another way of taking Oriental medicine. It is an important pathway but not available in common clinical situation. This experiment was performed in order to study the inhibitory effect of immediate-type allergic reaction of OG by anal therapy. In addition, the experiment was practised by 1 H-NMR spectroscopy to examine molecular structure of OG. The results were obtained as follows : OG concentration-dependently inhibited compound 48/80- induced immediate type systemic allergic reaction with concentrations of 0.01-1.0g/kg by anal administration 1 h before injection of compound 48/80. OG also concentration-dependently inhibited compound 48/80- induced ear swelling response with concentrations of 0.01-1g/kg by anal administration 1h before injection of compound 48/80. OG also inhibited the passive cutaneous anaphylaxis activated by anti-dinitrophenyl (DNP) IgE antibody concentration- dependently at concentrations ranging from 0.01 to 1g/kg. When OG was pretreated at concentrations ranging from 0.01 to 1g/ℓ, the histamine release from the rat peritoneal mast cells induced by compound 48/80 was reduced in a concentration-dependent manner. OG (0.1-1g/ℓ) had a significant inhibitory effect on histamine release from IgE-induced activated mast cells. OG is seen to be a biochemical compound certainly by 1 H-NMR spectroscopy According to above results, anal therapy of OG may be beneficial in the treatment of systemic and local immediate-type allergic reactions by inhibition of histamine release from mast cells.

Effects of Phyto-Extract Mixture (PEM381) in Type I Allergic Reaction-Induced Mice (제 I 형 알레르기 반응이 유도된 생쥐에서 식물추출 복합물(PEM381)의 효과)

  • Kim, Kyung-Bum;Park, Jeong-Su;Jeong, Jong-Moon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.11
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    • pp.1377-1384
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    • 2007
  • A previous study suggested that the phyto-extract mixture (PEM381) containing Camellia sinensis (leaf), Psidium guajava (leaf), and Rosa hybrida (flower) inhibited not only arachidonic acid cascade-related enzymes (5-lipoxygenase and cyclooxygenase) in vitro but also degranulation and histamine release from rat peritoneal mast cells. In this study, the same PEM381 was used to investigate its inhibitory effects on the syntheses of leukotrienes and prostaglandins as well as on serum concentration of histamine and eosinophils infiltration in type I allergic reaction-induced mice. The PEM381 could decrease concentrations of serum cysteinyl leukotrienes from mice activated by anti-DNP IgE and DNP-albumin. The concentration of serum histamine by oral administration of PEM381 (25, 50, and 100 mg/kg of body weight) in type I allergic reaction-induced mice was $395.93{\pm}190.37$ nM, $315.59{\pm}164.23$ nM, and $325.07{\pm}112.02$ nM, respectively, while that of positive control (promethazine hydrochloride 10 mg/kg of body weight) was $270.12{\pm}24.02$ nM. In addition, the PEM381 also showed inhibitory effect on the eosinophils infiltration in the nasal mucosa of mice which were sensitized with ovalbumin. However, the effect of PEM381 on the syntheses of prostaglandins seemed to be insignificant. Consequently, these results suggest that PEM381 may be useful for the prevention and treatment of type I allergy-related diseases.

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
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    • v.25 no.3
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    • pp.1-12
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    • 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.

Effect of Perillae Folium Extract on Regulation of Type 1 Allergic Response in RBL-2H3 Cells (자소엽(紫蘇葉) 에탄올 추출물이 RBL-2H3 비만세포에서 제 1형 알레르기 반응 조절에 미치는 효과)

  • Gok, Su-Yeong;Yu, Sun-Ae;Lee, Seung-Yeon
    • The Journal of Pediatrics of Korean Medicine
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    • v.26 no.1
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    • pp.36-45
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    • 2012
  • Objectives Perillae Folium (PF) has been widely used in Korean herbal medicines used for treatment of acute and chronic inflammatory diseases, such as rhinitis, asthma, and enteritis. In this study, to investigate the protective effect of PF on type 1 allergic response, we determined whether PF inhibits early or late allergic responses. Methods The effect of PF was analyzed by ELISA,. RT-PCR and Western blot 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 cytokines and enzymes were analyzed with RT-PCR. Signal transduction was analyzed with Western blot. Results We found that PF suppressed ${\beta}$-hexosaminidase release in RBL-2H3 by the IgE-DNP-HSA stimulation. PF also significantly inhibited enzymes level, such as COX-1, COX-2, iNOS, and HDC2, along with reduced cytokine levels, such as IL-2, IL-3, IL-4, IL-6, IL-13, and TNF-${\alpha}$ in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, and $I{\kappa}B{\alpha}$. Conclusions Our results indicate that PF protects against type 1 allergic response and exert an anti-inflammatory effect through the inhibition of degranulation and expression of cytokines and enzymes via the suppression of signal transduction.

Effect of Whitmania pigra whitman on the Allergic Inflammatory Response (수질(水蛭)이 비만세포의 알러지 염증 반응에 미치는 영향)

  • Lee, Ju-Yong;Kim, Eun-Kyoung;Oh, Hyun-A;Lee, Hyun-Sam;Sohn, Young-Joo;Jung, Hyuk-Sang;Kim, Yoon-Bum;Park, Seong-Kyu;Sohn, Nak-Won
    • The Journal of Korean Medicine
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    • v.29 no.2
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    • pp.81-95
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    • 2008
  • Objectives: Whitmania pigra whitman (WP) has been used in herbal medicine to treat various conditions, such as eczema, skin burns and frostbites in herbal medicine. The purpose of this study is was to investigate the effect of WP on anti-allergy mechanism. Methods: To clarify the mechanism, the effect of WP on vascular permeability of rat cutaneous tissue and histamine and cytokines (IL-6, IL-8, $TNF-{\alpha}$) released from mast cells were observed. Results: The results were 1. the pretreatment with WP significantly decreased the compound 48/80-induced degranulation and histamine release from RPMC 2. WP did not inhibit the anti-DNP IgE-induced increment of vascular permeability of rat cutaneous tissue 3. WP significantly reduced the PMA plus A23187-induced increment of expression of IL-6, IL-8, and $TNF-{\alpha}$ in HMC-1 cells. Conclusions: The present study providesevidence that WP acid inhibits mast cell-derived inflammatory allergic reactions by blocking histamine release and pro-inflammatory cytokine expression, and suggest the mechanisms of action. Furthermore, in vivo and in vitro anti-allergic effect of WP suggests a possible therapeutic application of this agent in inflammatory allergic diseases.

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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.

Anti-allergic and Anti-inflammatory Effect of Leonurus sibiricus Seed Ethyl Acetate Fractions (충울자 EtOAc 분획의 항알러지 및 항염증 효과)

  • Jung, Yujung;Jeon, Youngsic;Kim, Hyung Ja;Kang, Ki Sung;Kim, Yong Kee;Kim, Su-Nam
    • YAKHAK HOEJI
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    • v.58 no.5
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    • pp.294-299
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    • 2014
  • In this study, we investigate anti-allergic and anti-inflammatory effects of Leonurus sibiricus seed (LSS) extract in basophilic leukemia RBL-2H3 cells. To identify anti-allergic actions of LSS, the degranulation was evaluated in IgE and DNP-BSA stimulated RBL-2H3 cells. At the concentration of $100{\mu}g/ml$ of methanol (MeOH) extract and Methylene chloride (MC) and Ethyl acetate (EtOAc) fractions, the degranulation was significantly inhibited 16.7%, 16.7% and 27.9% respectively. And then, to assess anti-inflammatory effects of LSS, IL-4 and IL-13 mRNA level were detected in PMA/ionomycin (PI)-induced RBL-2H3 cells and cell proliferation and IL-4 mRNA level in isolated splenocytes from Balb/c mice. LSS MeOH extract and MC and EtOAc fractions significantly decreased the level of IL-4 and IL-13 mRNA in PI-induced RBL-2H3 cells and showed inhibitory effects on cell proliferation and expression of IL-4 mRNA level in mouse splenocytes. Taken together, these results suggest that LSS has potential anti-allergic and anti-inflammatory effects and EtOAc fraction is the most effective in regulating immune responses.