• Title/Summary/Keyword: degranulation

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A Murine Model of Toluene Diisocyanate-induced Contact Hypersensitivity

  • Chai, Ok Hee;Park, Sung Gil;Sohn, Jang Sihn;Hwang, Seung Soo;Li, Guang Zhao;Han, Eui-Hyeog;Kim, Hyoung Tae;Lee, Moo Sam;Lee, Hurn-Ku;Lee, Yong Chul;Song, Chang Ho
    • IMMUNE NETWORK
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    • v.2 no.3
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    • pp.158-165
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    • 2002
  • Background: Toluene diisocyanate (TDI) can cause contact allergy and occupational asthma, but the mechanism underlying sensitization to this chemical compound remains controversal. Also the correlation of mast cell with contact hypersensitivity (CHS) and the role of mast cell in the TDI-induced CHS is unknown. This issue was investigated by administrating TDI on the skin of genetically mast cell-deficient WBB6F1/$J-Kit^{W}/Kit^{W-v}$ ($W/W^{V}$) and congenic normal WBB6F1/J-Kit+/+ (+/+) mice. Methods: To development of animal model of TDI-induced CHS and to investigate the correlation of mast cell with CHS and the role of mast cell in the TDI-induced CHS, $W/W^V$ and +/+ mice were sensitized with TDI on the back skin at day 1 and day 8, and then challenged with 1% TDI on the ear at day 15. At 1, 2, 4, 8, and 24 hours after 1% TDI challenge, the ear thicknesses were measured. It was investigated the histologic changes of dermis in the ear of $W/W^V$ and +/+ mice at 24 hours after 1% TDI challenge. Results: TDI induced a significant ear swelling response in $W/W^V$ and +/+ mice. TDI induced the significant infiltrations of polymorphonuclear leukocytes and eosinophils in $W/W^V$ and +/+ mice, but not of mast cells in normal mice. And TDI increased a characteristic extent of mast cell degranulation in normal mice. There were no significant differences in the ear swelling and the infiltrations of polymorphonuclear leukocytes and eosinophils of normal versus $W/W^V$ mice, either at baseline or after TDI-induced CHS. Conclusion: From the above results, TDI can be used as a murine CHS model, and the mast cells may not be essential in TDI-induced CHS.

Comparison of Allergic Parameters between Whey Protein Concentrate and Its Hydrolysate in Rat Basophilic Leukemia (RBL)-2H3 Cells

  • Kim, Hana;Ahn, Sung-Il;Jhoo, Jin-Woo;Kim, Gur-Yoo
    • Food Science of Animal Resources
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    • v.38 no.4
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    • pp.780-793
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    • 2018
  • This study was conducted to compare the anti-allergic effects of a whey protein concentrate (WPC) and WPC hydrolysate. WPC hydrolysate was prepared using enzymatic digestion for 8 h with trypsin and ${\alpha}$-chymotrypsin, after which it was freeze-dried. The allergic parameters assessed in rat basophilic leukemia (RBL)-2H3 cells were degranulation and release of ${\beta}$-hexosaminidase, release of tumor necrosis factor $(TNF)-{\alpha}$, and changes in the expression of $IL-1{\beta}$, IL-4, and IL-10 by real time polymerase chain reaction (PCR). During preparation of the WPC hydrolysate, hydrolysis increased rapidly from 0 to 10 min and then gradually increased slowly from 1 h onwards, achieving a final degree of hydrolysis of 78.50%. The SDS-PAGE analysis revealed a reduction in the intensity of several protein bands in the WPC hydrolysate compared to the WPC. IgE-induced ${\beta}$-hexosaminidase release from RBL-2H3 cells was decreased to a higher degree following treatment with the hydrolysate compared to WPC treatment. W500 ($500{\mu}g/mL$ WPC) showed the least inhibition of ${\beta}$-hexosaminidase release, but there was no significant difference between W500 and W1000 ($1,000{\mu}g/mL$) (p<0.05). H1000 ($1,000{\mu}g/mL$ WPC hydrolysate) inhibited ${\beta}$-hexosaminidase release by 39%. Compared to the control, treatment with H1000 decreased $TNF-{\alpha}$ secretion to 11.87 pg/mL. The gene expression levels of IL-1${\beta}$, IL-4, and IL-13 were all significantly decreased in hydrolysate (p<0.05). In the case of $IL-1{\beta}$ and IL-4, the expression levels in W1000 treated cells were decreased by 73.67% and 65%, respectively, and that of IL-13 was decreased by 66.43% compared to the control.

The Experimental Study on the Anti-allergic Effects of Bojungikgi-tang (보중익기탕이 알레르기반응에 미치는 영향에 대한 실험적연구)

  • 강대성;감철우;박동일
    • Journal of Life Science
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    • v.13 no.1
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    • pp.73-82
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    • 2003
  • This experimental research has been done to study the effects of Bojungikgi-tang(BJIKT) on the anti-allergic reaction. We found the several important results from the research which has been performed by two experiments toward immediately type and delayed type in order to study the effects of BJIKT on hypersensitivity response to mice. The results obtained from our research are as following: 1. The survival rate of one group to which we injected only the compound 48/80 is almost 0% according to its density and timing test. In the other hand, the survival rates of the other group to which we injected both of the compound 48/80 and BJIKT are 20%, 10%, 30%, 10%, 40%, and 70% according to 0.025, 0.05, 0.1, 0.25 0.5 and 1(mg/g) of compound 48/80. Time dependency test also shows the 10% and 0% survival rates in 5 and 10 minutes. 2. BJIKT revealed the significantly inhibitory effect on Compound 48/80 induced Mast cell degranulation. 3. BJIKT showed the significantly inhibitory effect in the delayed type hypersensitivity response to picryl chloride. 4. BJIKT showed the significantly inhibitory effect in the delayed type hypersensitivity response to sheep red blood tell. Our research provides the important evidence that BJIKT is benificial to the prevention and treatment of allergy related diseases.

Anti-Allergic Effect of 1,2,3,4,6-Penta-O-Galloyl-β-D-Glucose on RBL-2H3 Cells (RBL-2H3 세포에서 1,2,3,4,6-Penta-O-Galloyl-β-D-Glucose의 항알레르기 효과)

  • Kim, Yoon Hee;Choi, Ye Rang;Kim, Ji Young;Kwak, Sang Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.613-618
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    • 2016
  • 1,2,3,4,6-Penta-O-galloyl-${\beta}$-D-glucose (PGG) is a gallotannin isolated from various plants such as Galla Rhois. In a previous study, it was reported that PGG has anti-allergic effects by inhibiting interleukin (IL)-4 signaling in B cells. However, the effect of PGG on basophilic cells remains unclear. Therefore, the aim of this study was to investigate the inhibitory effect of PGG on mitogen and calcium ionophore-induced allergic responses. PGG had no effect on proliferation and cytotoxicity of RBL-2H3 cells. PGG significantly suppressed cell degranulation (histamine and ${\beta}-hexosaminidase$) as well as inflammatory cytokine production such as IL-4 and tumor necrosis factor-${\alpha}$. The underlying mechanism of PGG on these anti-allergic actions was correlated with inhibition on translocation of nuclear factor-${\kappa}B$ from the cytosol to nucleus. These data suggest that PGG is a potentially effective functional compound for prevention of allergic diseases.

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.

Antioxidative and Anti-asthma Effect of Morus Bark Water Extracts (상백피 물 추출물의 항산화 활성 및 항천식 효과)

  • Kim, Jeong-Mi;Baek, Jong-Mi;Kim, Hyun-Sook;Choe, Myeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.9
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    • pp.1263-1269
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    • 2010
  • This study was performed to evaluate the antioxidant activities and anti-asthma effects of Morus bark water extracts. Inhibitory effect of Morus bark onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activities in vitro. Anti-asthma activities of Morus bark water extracts were assessed by testing their effects on the degranulation of mast cell. For this, $\beta$-hexosaminidase released from a basophilic cell line, RBL-2H3 was used and pro-inflammatory cytokines were measured by ELISA kit. The antioxidant activities of water extracts of Morus bark was 59.2% in the DPPH assay at $2,000\;{\mu}g/mL$ and 78.8% in the hydroxyl radical scavenging assay at $2,000\;{\mu}g/mL$. Our results indicated that Morus bark water extracts effectively inhibited free radical generation. Morus bark water extracts inhibited inflammation-mediating substances such as histamine and $\beta$-hexosaminidase release from RBL-2H3 cells. Cytokine release demonstrated a more effective blockading ability of the Morus bark water extracts to the release of IL-4 and TNF-$\alpha$ compared to control. These results demonstrate that Morus bark may be beneficial in the treatment of allergic inflammatory disease.

Rehmannia Glutinosa Pharmacopuncture Solution Regulates Functional Activation, FcεRI Expression, and Signaling Events in Mast Cells

  • Kang, Kyung-Hwa;Lee, Kyung-Hee;Yoon, Hyun-Min;Jang, Kyung-Jeon;Song, Chun-Ho;Kim, Cheol-Hong
    • Journal of Pharmacopuncture
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    • v.15 no.4
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    • pp.32-41
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    • 2012
  • Objectives: Rehmannia glutinosa pharmacopuncture solution (RGPS) was investigated to determine both its anti-allergic inflammatory effects on mast cells and its detailed mechanism of actions. Methods: We investigated whether RGPS suppress cytokines, enzymes, $Fc{\varepsilon}RI$ expression and $Fc{\varepsilon}RI$-mediated signaling in RBL-2H3 cells stimulated with anti-DNP IgE/DNP-HSA. The suppressive effects of RGPS on the levels of cytokines such as IL-$1{\beta}$, IL-6 and GM-CSF were measured using emzyme-linked immunospecific assay (ELISA). The mRNA expression levels of cytokines, enzymes (HDC2, COX-1, COX-2 and 5LO) and $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunits were measured using reverse transcription polymerase chain reaction (RT-PCR) method. The activation of $Fc{\varepsilon}RI$-mediated signaling was examined using Western blot analyses. Results: RGPS suppressed production of proinflammatory cytokines (IL-$1{\beta}$, IL-6, and GM-CSF) in stimulated RBL-2H3 cells significantly (p < 0.05). RGPS also suppressed mRNA expression of inflammatory enzymes (HDC2, COX-1, COX-2, 5LO). In addition, mRNA expression levels of $Fc{\varepsilon}RI{\alpha}$, $Fc{\varepsilon}RI{\beta}$and $Fc{\varepsilon}RI{\gamma}$ were lowered by treatment with RGPS. Finally, RGPS prevented phosphrylation of Lyn, Syk, LAT, Gab2, PLC ${\gamma}1/2$, PI3K, Akt, cPLA2 and $I{\kappa}B{\alpha}$. Conclusions: RGPS effectively suppresses mast cell activations such as degranulation and inflammatory response via down-regulation of the $Fc{\varepsilon}RI$-mediated signaling pathways in IgE/Ag-stimulated mast cells.

Anti-inflammatory Effects of Quercetin and Vitexin on Activated Human Peripheral Blood Neutrophils - The effects of quercetin and vitexin on human neutrophils -

  • Nikfarjam, Bahareh Abd;Hajiali, Farid;Adineh, Mohtaram;Nassiri-Asl, Marjan
    • Journal of Pharmacopuncture
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    • v.20 no.2
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    • pp.127-131
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    • 2017
  • Objectives: Polymorphonuclear neutrophils (PMNs) constitute the first line of defense against invading microbial pathogens. Early events in inflammation involve the recruitment of neutrophils to the site of injury or damage where changes in intracellular calcium can cause the activation of pro-inflammatory mediators from neutrophils including superoxide generation, degranulation and release of myeloperoxidase (MPO), productions of interleukin (IL)-8 and tumor necrosis factor ${\alpha}$ ($TNF-{\alpha}$), and adhesion to the vascular endothelium. To address the anti-inflammatory role of flavonoids, in the present study, we investigated the effects of the flavonoids quercetin and vitexin on the stimulus-induced nitric oxide (NO), $TNF-{\alpha}$, and MPO productions in human neutrophils. Methods: Human peripheral blood neutrophils were isolated, and their viabilities were determined by using the Trypan Blue exclusion test. The polymorphonuclear leukocyte (PMNL) preparations contained more than 98% neutrophils as determined by morphological examination with Giemsa staining. The viabilities of cultured neutrophils with various concentrations of quercetin and vitexin ($1-100{\mu}M$) were studied using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without quercetin and vitexin ($25{\mu}M$) for 45 min, and stimulated with phorbol 12-myristate 13-acetate (PMA) ($10^{-7}M$). NO production was carried out through nitrite determination by using the Griess method. Also, the $TNF-{\alpha}$ and the MPO productions were measured using enzyme-linked immunosorbent assay (ELISA) kits and MPO assay kits. Results: Neutrophil viability was not affected up to a concentration of $100{\mu}M$ of quercetin or vitexin. Both quercetin and vitexin significantly inhibited $TNF-{\alpha}$, NO, and MPO productions in human neutrophils (P < 0.001). Conclusion:The present study showed that both quercetin and vitexin had significant anti-inflammatory effects. Thus, treatment with either quercetin or vitexin may be considered as a therapeutic strategy for treating patients with neutrophil-mediated inflammatory diseases.

Effects of Mahwangyoonpye-tang on Asthma Induced by Ovalbumin in Mouse (마황윤폐탕이 ovalbumin으로 유발된 mouse의 천식에 미치는 영향)

  • Kim Ji Yun;Park Dong il;Kim Jong Dae;Kam Cheol Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.6
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    • pp.1453-1462
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    • 2003
  • In order to study the effect of oral administration of Mawhangyounpye-tang against to asthma, astham was induced to allergy-sensitive Balb/c mouse with ovalbumin using method of Hatfield et al (1997). The changes of diameter lumen of upper portion of the trachea, lung weight, gross appearance of lung, histological profiles of lung and trachea, numbers of cellular compartments in the bronchoalveolar lavage fluid (BALF), numbers and morphology of the mast cells in the trachea, numbers of mucus-secretory cell in the broncus, morphology of the bronchus, ultramicroscopical appearance of surface of trachea and number of cilia and mucous-secretory cells by scanning electron microscope. Obtained results were as follows. 1. The diameters of trachea lumen were significantly decreased in asthma induced control groups and these decreasing were result from hypertrophy of mucous membrane. However, these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 2. Lung weights and black spots, which were result from infiltration of inflammatory cells, were significantly increased in asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 3. Hypertrophy of mucous membrane of trachea and bronchus and !bronchioles in the lung, peritracheal, peribronchus and peribronchiolar inflammatory cell infiltration, and mucoid exudate deposit in the lumen were observed in asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 4. Cellular compartments including neutrophil and eosinophil were dramatically increased in the BALF of asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 5. Mast cell degranulation and decreasing of the numbers of mast cells were detected in the trachea of asthma induced control groups. However, these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. 6. Shed, decreasing of cilia cell and increasing of mucous-secretory cells in the surface of the trachea of asthma induced control groups but these phenomena were dramatically recovered in the Mawhangyounpye-tang dosing groups. In conclusion, it Is considered that Mawhangyounpye-tang has somewhat favorable effect on the asthma because the asthma specific series of abnormalities in respiratory system were decreased after oral administratin of Mawhangyounpye-tang in this study. In future, it is needed that the toxicological and dosagespecific study of Mawhangyounpye-tang to use against asthma with safe.

Effects of 0.1% Tacrolimus on Canine Skin Mast Cells and Eosinophils (개에서 0.1% FK-506 연고의 피부 비만세포와 호산구에 미치는 영향)

  • Jeong, A Young;Bae, Seulgi;Oh, Taeho
    • Journal of Veterinary Clinics
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    • v.31 no.2
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    • pp.95-101
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    • 2014
  • Five dogs were used to determine whether 0.1% tacrolimus ointment application for one day would inhibit IgE-mediated late-phase reactions (LPRs). It was consisted of three periods: one period without therapeutic administration (control) and two periods of treatment with either the tacrolimus ointment or vehicle. Induction of IgE-mediated LPRs was induced by intradermal injections of 0.05 ml (0.14 mg/ml) of solution of goat anti-canine IgE polyclonal antibodies. Each section for mast cells (MCs) and eosinophils (EPs) was stained with acidified toluidine blue, and Luna's stain, respectively. Assessment of anti-inflammatory effect of tacrolimus ointment composed of cell counts of MC and EP from lesions of induced LPR. In normal canine biopsies, the number of dermal MCs and EPs were $12.3{\pm}1.4cells/mm^2$ and $3.1{\pm}1.3cells/mm^2$, respectively. MC counts dramatically decreased at time dependent manner after anti-IgE administration. However, the number of MCs on 6 hours after challenge was significantly less decreased in the groups treated with the tacrolimus, as compared with control and vehicle group. The number of EPs on 24 hours after challenge was significantly lower in the group treated with the tacrolimus than in the control and vehicle groups. In conclusion, this study revealed that 0.1% tacrolimus ointment in dogs may exert a potent anti-inflammatory effect on inhibition of MC degranulation and also secondary prevention of EP infiltration during LPR.