• The Korea Journal of Herbology
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• v.28 no.1
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• pp.33-42
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• 2013

Objective : Morus alba Linne Root Bark (MRAL) is a medicinal herb in Korean Medicine, known for its anti-inflammatory and anti-allergic properties. However, its mechanisms of action and the cellular targets have not yet been found and the study was developed to investigate the allergic suppressive effect of MRAL. The purpose of this study is to investigate the allergic suppressive effects of MRAL on activation of MC/9 mast cells. Methods : Cytotoxic activity of MRAL (50, 100, 200, 400 ${\mu}g/mL$) on MC/9 mast cells measured using EZ-Cytox cell viability assay kit (WST reagent). The levels of interleukin-5 (IL-5), IL-13 and IL-4, IL-5, IL-6, IL-13 mRNA expression were measured by enzyme-linked immunosorbent assay (ELISA) and real-time PCR respectively. The expression of transcription factors such as GATA-1, GATA-2, NFAT, AP-1 and NF-${\kappa}B$ p65 DNA binding activity were measured by western blot and electrophoresis mobility shift assay (EMSA). Results : Our results indicated that MRAL (50 ${\mu}g/mL$, 100 ${\mu}g/mL$) significantly inhibited PMA/Ionomycin-induced production of IL-5 and IL-13 and the expression of IL-4, IL-5, IL-6 and IL-13 mRNA in MC/9 mast cells. Moreover, MRAL (50 ${\mu}g/mL$, 100 ${\mu}g/mL$) inhibited PMA/Ionomycin-induced GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos protein expression and NF-${\kappa}B$ p65 DNA binding activity in MC/9 mast cells. Conclusions : In conclusion, we suspect the anti-allergenic activities of MRAL, may be related to the regulation of transcription factors GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-${\kappa}B$ p65 DNA binding assay causing inhibition of Th2 cytokines IL-5 and IL-13 in mast cells.

• The Korea Journal of Herbology
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• v.26 no.4
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• pp.1-7
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• 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.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.1
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• pp.33-45
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• 2009

Objective : Lacca Sinic: Exsiccate (LSE) extracted from Rhus vemicitlus Stokes (RVS) has been used traditionally as a remedy for inflammation in Korea, China, and Japan. However, as yet there is no clear explanation of how LSE affects the production of inflammatory cytokines. This study was to determine the effects of LSE on the mast cell-mediated inflammatory responses. Method : We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A231S7) in the human mast cell line (HMC-l) incubated with various concentrations of Laces Sinica Exsiccate (LSE). The $TNF-\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The $TNF-\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. Nuclear and cytoplasmic proteins were examined by Western blot analysis. The NF-${\kappa}B$ promoter activity was examined by a luciferase assay. Result : LSE inhibited the PMA + A231S7-induced $TNF-\alpha$, IL-6, and IL-8 expression and suppressed NF-${\kappa}B$ activation in the stimulated-HMC-1. In addition, LSE inhibited induction of NF-${\kappa}B$ promoter-mediated luciferase activity. Conclusion : In this study, we have found that LSE is an inhibitor of NF-${\kappa}B$ and cytokines on the mast cell-mediated inflammatory responses.

• Korean Journal of Plant Resources
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• v.24 no.3
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• pp.324-329
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• 2011

Eleutherococcus senticosus Maxim. has been successfully used as an oriental medicine for various diseases including allergic disorders. Histamine is a major factor on various allergic responses and it is reported that histamine was released from mast cells by sensitization of allergens. In this study, ethanol extracts from E. senticosus Maxim. were prepared and the composition was analyzed by high performance liquid chromatography. The eleutheroside B as a primary effective component of E. senticosus was contained approximately 225 mg/kg in root bark extracts. The extracts were found to significantly inhibit compound 48/80-induced histamine release form mast cells in dose dependent manner. However the extracts had low cytotoxicity on the mast cells with MTT assay. These results showed that E. senticosus Maxim. extracts may be the effective materials on inflammatory disorders.

• The Journal of Internal Korean Medicine
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• v.11 no.2
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• pp.22-42
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• 1990

Sabaiksan has been prescribed to treat various allergic diseases in herbal medicine which were induced by various vasoactive amine released from the mast cells. The constituents of Sabaiksan are Mori Cortex Radices(MCR), Lycii Cortex Radicis(LCR) and Glycyrrhizae Radix(GR). Recently, simple models of compound 48/80 induced anaphylactic shock and cutaneous reaction in vivo were developed to test various agents employed in the field of allergy and toxicology research. The purpose of this study is to evaluate the effects of Sabaiksan on compound 48/80 induced anaphylactic stock, cutaneous reaction and mesenteric mast cell degranulation rate in ICR mice, and on compound 48/80 induced peritoneal mast cell degranulation and histamine release in vitro. Groups of ICR mice were intraperitoneally pretreated with $100{\mu}{\ell}$ of saline, $MCR(2g/m{\ell}),\;LCR(2g/m{\ell}),\;GR(g/m{\ell})$ or Sabaiksan itself(MCR+LCR+GR) at 24, 12 and 1 hour before compound 48/80 solution ($10{\mu}{\ell}/gm$ B. W) were peritoneally given into them, and then mortality within 72 hours after the compound 48/80 injection, and mesenteric mast cell degranulation rate at 15 minutes after compound 48/80 injection were calculated. In vitro experiment, $400{\mu}{\ell}$ of rat peritoneal mast cell suspension$(10^6cell/m{\ell})$ were pretreated with $50{\mu}{\ell}$ of saline, $MCR(2g/m{\ell}),\;LCR(2g/m{\ell}),\;GR(g/m{\ell})$ or Sabaiksan itself at room temperature for 30 minutes, and then $50{\mu}{\ell}$ of compound 48/80 solution $(100{\mu}g/m{\ell})$ were added into it. 30 minutes after the addition of compound 48/80 solution, histamine release assay in the supernatant of peritoneal mast cell suspension were performed employing radioisotope enzymatic assay and morphologic changes of mast cells in each regular time point were photographed. Compared with controls, compound 48/80 induced anaphylactic shock was significantly inhibited by MCR and GR pretreatment into the ICR mice. Significant inhibition of compound 48/80 induced cutaneous reaction, mesenteric mast cell degranulation rate in vivo and histamine release from the rat peritoneal mast cells in vitro was observed only in MCR pretreated group. From the above results, it is suggested that MCR component of Sabaiksan may playa key role to suppress mast cell function since it has been applied to various allergic diseases.

• 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.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.282-290
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• 2019

Background: Ginsenoside Rg3 (G-Rg3) is the major bioactive ingredient of Panax ginseng and has many pharmacological effects, including antiadipogenic, antiviral, and anticancer effects. However, the effect of G-Rg3 on mast cell-mediated allergic inflammation has not been investigated. Method: The antiallergic effects of G-Rg3 on allergic inflammation were evaluated using the human and rat mast cell lines HMC-1 and RBL-2H3. Antiallergic effects of G-Rg3 were detected by measuring cyclic adenosine monophosphate (cAMP), detecting calcium influx, and using real-time reverse transcription polymerase chain reaction, enzyme-linked immunosorbent assay, Western blotting, and in vivo experiments. Results: G-Rg3 decreased histamine release from activated mast cells by enhancing cAMP levels and calcium influx. Proinflammatory cytokine production was suppressed by G-Rg3 treatment via regulation of the mitogen-activated protein kinases/nuclear factor-kappa B and receptor-interacting protein kinase 2 (RIP2)/caspase-1 signaling pathway in mast cells. Moreover, G-Rg3 protected mice against the IgE-mediated passive cutaneous anaphylaxis reaction and compound 48/80-induced anaphylactic shock. Conclusion: G-Rg3 may serve as an alternative therapeutic agent for improving allergic inflammatory disorders.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.22 no.2
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• pp.60-73
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• 2009

Objective : Houttuyniae Herba is widely used in oriental medicine as a remedy for inflammation. However, as yet there is no clear explanation of how Houttuyniae Herba affects the production of inflammatory cytokine. This study was to determine the effects of Essence extracted from Houttuynia cordata Thunb(HCT) on the mast cell-mediated inflammatory responses. Method : We measured the amount of inflammatory cytokine production induced by the phorbol myristate acetate (PMA) plus calcium ionophore(A23187) in the human mast cell line (HMC-1) incubated with various concentrations of HCT. The TNF-$\alpha$, IL-6 and IL-8 secreted protein levels were measured by the ELISA assay. The TNF-$\alpha$, IL-6 and IL-8 mRNA levels were measured by the RT-PCR analysis. Nuclear and cytoplasmic proteins were exmined by Western blot analysis. The NF-$\kappa$B promoter activity was examined by a luciferase assay. Result : HCT inhibited the PMA + A23187-induced TNF-$\alpha$, IL-6 expression and reduced mRNA of TNF-$\alpha$, IL-6 and IL-8. we observed that HCT suppressed the induction of NF-B activity. In addition, HCT suppressed PMA plus A23187-induced NF-$\kappa$B promoting activity. Conclusion : In this study, we have found that HCT is an inhibitor of NF-$\kappa$B and cytokines on the mast cell-mediated inflammatory responses.

• The Korea Journal of Herbology
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• v.23 no.4
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• pp.81-89
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• 2008

Objectives: In this study, the pharmacological effects of the ethylacetate extract of Ostericum koreanum(North Kangwhal; NK) on allergic inflammation were investigated in activated human mast cells. Methods: North Kangwhal was extracted with 80% methanol for 24 h, and then fractionated with ethylacetate(NK-EtOAc extract). HMC-1 cells, an human mast line, were pre-incubated with different concentrations of NK-EtOAc extract for 30 min, and then stimulated with PMA(50 nM/ml) and A23187($1{\mu}M/ml$) at indicated times. The cell toxicity was determined by MTT assay. The concentrations of prostaglandin E2(PGE2) and cytokines(TNF-${\alpha}$, IL-8) were measured by enzyme-linked immunosorbant assay. Results: NK-EtOAc extract($10{\sim}50{\mu}g/ml$) significantly inhibited the productions of $PGE_2$, TNF-${\alpha}$ and IL-8 in PMA/A23187-stimulated HMC-1 cells without cell toxicity($0{\sim}50{\mu}g/ml$). NK-EtOAc extract also inhibited PMA/A23187-induced phosphorylation of ERK1/2 MAPK and the NF-${\kappa}B$ p65 subunit translocation into the nuclear of HMC-1 cells. Conclusions: This study suggests that NK-EtOAc extract may have an anti-inflammatory property through suppressing the production of inflammatory mediators in activated mast cells and its molecular mechanism underlies the blocking of NF-${\kappa}B$ pathway.

• The Korea Journal of Herbology
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• v.32 no.6
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• pp.9-15
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• 2017

Objectives : Traditional medicines isolated from natural products often have positive effects in the prevention and healing of various immune disorders, such as allergy and atopic inflammation. Scrophulariae Radix (SR) been used in oriental medicine used for treatment of acute and chronic inflammatory diseases. Mast cells are known to play important roles in the initiation of allergic reactions. In this study, we investigated the effects of SR ethanol extract on inflammatory responses in IgE-stimulated RBL-2H3 mast cells. Methods : Rat basophilic leukemia RBL-2H3 cells were purchased from Korean Cell Line Bank (KCLB No. 22256). Cell viability was measured by MTT assay. Assays for ${\beta}-Hexosaminidase$ Secretion : RBL-2H3 cells were sensitized with dinitrophenyl-ImmunoglobulinE (DNP IgE). The next antigen DNP-BSA ($25ng/m{\ell}$) was added for 10 minutes and the reaction was terminated after 5 minutes in the ice bath. To determine ${\beta}-Hexosaminidase$ release, supernatants were aliquoted into 96-well plates. Samples were mixed with substrate solution and incubated for 1 h at $37^{\circ}C$. Absorbance was measured with a spectrophotometer at 405 nm. IL-4 and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$) concentrations in cell culture supernatants were measured using enzyme-linked immunosorbent assay (ELISA) kits. Results : The cytotoxicity of SRE in RBL-2H3 cells was less than 5%. SRE inhibited DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. Also significantly decreased the levels of inflammatory cytokine, IL-4 and TNF-alpha. In this study, the SRE showed potential anti-allergic and antiinflammatory. Conclusions : These results indicate that SRE could be inhibit the allergic response through suppressing the mast cell activation.