• 한국환경과학회지
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• 제26권12호
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• pp.1355-1362
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• 2017

This study was performed to evaluate the possibility of application of lactic acid bacteria fermentation to increase the anti-allergic activity of the extracts from Houttuynia cordata Thunb. H. cordata Thunb was fermented on 25, 30, 35 and $40^{\circ}C$ for 5 days by two species of lactic acid bacteria, Leuconostoc mesenteroides 4395 and Lactobacillus sakei 383. The anti-allergic activity of water extracts of H. cordata Thunb was then analyzed both before and after fermentation. Anti-allergic activity was determined in vitro assays by using 5-lipoxygenase (5-LO), cyclooxygenase-2 (COX-2) and ${\beta}$-hexoseaminidase release of RBL-2H3 cells (degranulation marker). The extracts fermented at $35^{\circ}C$ by both bacteria had the highest inhibitory activities against 5-LO, and also higher than the control, and the one fermented at $30^{\circ}C$ by both bacteria had the highest inhibitory activity against COX-2. The degranulation of RBL-2H3 cells induced by IgE-antigen complex was estimated as ${\beta}$-hexoseaminidase release rate as reference of 100%, the release rates were inhibited in $25{\mu}g/ml$ of the extracts fermented at 30, 35 and $40^{\circ}C$ only by L. mesenteroides 4395. These results indicate that anti-allergic activity of H. cordata Thunb is increased by lactic acid bacteria fermentation.

• Journal of Nutrition and Health
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• 제43권4호
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• pp.367-373
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• 2010

RBL-2H3 세포에서 차조기씨 물 추출물의 항산화성 및 항염증 효과를 알아보기 위하여 DNP-IgE와 HSA로 세포를 활성화시켰다. 활성화된 비만 세포에서 분비되어 나오는 히스타민을 효소적 방법으로 측정하였으며, 염증성 사이토카인 IL-4과 TNF-$\alpha$의 생성량은 ELISA 방법으로 측정하였다. 차조기씨 물 추출물은 농도 의존적으로 항산화 활성이 증가되었으며, $1,000\;{\mu}g$/mL농도에서 DPPH와 hydroxyl radical 소거능이 각각 38.9, 93.1%로 가장 높게 나타났다. 이는 차조기씨에 함유된 130.7 mg/g의 높은 페놀 함량에 기인하여 항산화 효과를 나타내었다. 한편 차조기씨 물 추출물을 처리하지 않은 세포에서는 현저히 높은 양의 히스타민을 분비하였지만, 각 추출물의 농도로 처리시킨 세포에서의 분비량은 유의적으로 감소되었고, IL-4와 TNF-$\alpha$의 분비량 모두 차조기씨 물 추출물을 처리군은 대조군에 비해서 유의적으로 낮은 생성량을 보여주었다. 결과적으로 차조기씨 물추출물은 비만세포에 항원-항체 반응으로 인해 발병되는 제1형 알레르기 반응의 약학적 효능을 가진 것으로 나타나 임상적으로 유용하게 사용될 수 있을 것으로 사용되며, 향후 이에 대한 심층적이고 체계적인 연구가 뒤따라야 할 것으로 생각된다.

• 동의생리병리학회지
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• 제17권2호
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• pp.428-435
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• 2003

This research was performed to examine an anti-inflammatory effects of Gamisangryosamul-Tang(GSS) and anti-pruritus effects of Ziyang-Go(Salve). This study was processed by three experiments; Experiment 1: Inhibitory activity of GSS extract on the degranulation of mast cell and histamine release in plasma induced by compound 48/80 i.p, injection after the pretreatment of GSS extract i.p, injection in Sprague-Dawley rats, Experiment 2: Anti-inflammatory effect of GSS extract on macropharge raw 264,7 cells treated by LPS 250 ppm (before 2 hours). Experiment 3: Measurement of passive cutaneous anaphylaxis and atopic dermatitis using NC/Nga mice, GSS extract inhibited histamine release by 70% compared to compound 48/80 treated control group and histologically significantly reduced (P<0,01) the degranulation of mast cell in SD rats. In GSS extract treated group, the expression of TNF-α in macropharge cell showed the remarkable inhibitory effect about 62% (P<0,01) compared to LPS treated control group. The expression of IL-6 appeared more effective by 46% than the LPS treated control group and by 6% compared to hydrocortison treated group, Comparing with steroid (0.05% prednisolon) ointment, Ziyan-Go treated group showed the significant(30%) recovery on skin response index in atopic dermatis like anaphylaxis mice(NC/Nga), Finally, in scratching behavioral tests of NC/Nga mice for three weeks, Ziyang-Go treated group significantly (P<0.05) suppressed the pruritus on the face, neck, ears and dorsal skin than inbred NC/Nga mice. However, the change of IgE and IFN-γ from the spleen cell of NC/Nga mice was not significantly different between the oral intake of GSS extract group and of saline intaked control group. Summary and Conclusion: This study demons trates that Ziyang-go have the equal anti-pruritus effect to steroid ointment and GSS extract have the notable immunologic activity on inflammatory in vivo and in vitro model. Advanced experiment of this study will be required for more reliable information about the correlation between the lymphokine (i.e. IgE) and the anti-allergic effects of GSS.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.456-464
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• 2020

Mast cells (MCs) are systemically distributed and secrete several allergic mediators such as histamine and leukotrienes to cause type I hypersensitivity. Dasatinib is a type of anti-cancer agent and it has also been reported to inhibit human basophils. However, dasatinib has not been reported for its inhibitory effects on MCs or type I hypersensitivity in mice. In this study, we examined the inhibitory effect of dasatinib on MCs and MC-mediated allergic response in vitro and in vivo. In vitro, dasatinib inhibited the degranulation of MCs by antigen stimulation in a dose-dependent manner (IC₅₀, ~34 nM for RBL-2H3 cells; ~52 nM for BMMCs) without any cytotoxicity. It also suppressed the secretion of inflammatory cytokines IL-4 and TNF-α by antigen stimulation. Furthermore, dasatinib inhibited MC-mediated passive cutaneous anaphylaxis (PCA) in mice (ED₅₀, ~29 mg/kg). Notably, dasatinib significantly suppressed the degranulation of MCs in the ear tissue. As the mechanism of its effect, dasatinib inhibited the activation of Syk and Syk-mediated downstream signaling proteins, LAT, PLCγ1, and three typical MAP kinases (Erk1/2, JNK, and p38), which are essential for the activation of MCs. Interestingly, in vitro tyrosine kinase assay, dasatinib directly inhibited the activities of Lyn and Fyn, the upstream tyrosine kinases of Syk in MCs. Taken together, dasatinib suppresses MCs and PCA in vitro and in vivo through the inhibition of Lyn and Fyn Src-family kinases. Therefore, we suggest the possibility of repositioning the anti-cancer drug dasatinib as a treatment for various MC-mediated type I hypersensitive diseases.

• 생명과학회지
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• 제17권3호통권83호
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• pp.350-355
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• 2007

흰쥐유래의 비만세포인 RBL-2H3 세포는 다양한 Src-family kinase를 발현한다. 현재까지의 연구결과에 의하면 비만세포의 초기 활성화에 Lyn kinase가 중요한 역할을 한다고 알려져 왔다. 그러나 그 세포에서 발현되는 다양한 다른 Src-family kinase의 역할은 불분명하다. 본 연구에서는 비만세포에서 다양한 Src-family kinase가 세포내 다른 곳에서 다양하게 발현되고 있다는 사실을 RT-PCR, immunoblotting 그리고 confocal microscopy 기법을 이용하여 증명하였다. 그 결과 Lyn 및 Fgr kinase는 세포막에 위치하고 c-Src 및 Yes kinase는 세포 내 과립에 존재하는 것을 알 수 있었다. 모든 Src-family kinase를 클로닝하고 과발현하여 탈과립 대한 영향을 평가하였다. 그 결과 fyn과 Fgr kinase는 비만세포에서 항원 유도의 탈과립을 증가시켰으며 반면 Lyn kinsae는 탈과립을 억제시키는 것을 확인할 수 있었다. 이러한 결과는 비만세포 초기 신호전달계에서 Fgr가 중요한 역할을 할 가능성을 제시한다.

• Journal of the Korean Wood Science and Technology
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• 제41권6호
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• pp.551-558
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• 2013

본 연구는 만병초와 구상나무 추출물의 항알러지 효과에 대해 조사하였다. 비만세포 RBL-2H3에 concanavalin A (Con A)를 처리하여 면역반응을 유도한 후 만병초와 구상나무 추출물을 농도별로 처리하여 real-time PCR을 이용하여 IL-4, IL-13의 mRNA 발현량을 확인하였다. 또한 ${\beta}$-hexosaminidase 분비를 측정하여 탈과립 유도효과를 평가하였다. IL-4의 발현은 만병초 $10^{-7}$, $10^{-5}$, $10^{-3}%$ 농도와 구상나무의 모든 농도에서 유의성 있게 감소하였으며, IL-13의 발현은 만병초와 구상나무 모든 농도에서 유의성 있게 감소되었다. 만병초 추출물과 구상나무 추출물의 $10^{-5}$, $10^{-3}%$ 농도에서는 ${\beta}$-hexosaminidase 분비가 유의성 있게 감소하였다. 이러한 결과를 통해 만병초와 구상나무 추출물이 면역반응 표적유전자의 발현을 감소시키고 면역반응에 의해 유도되는 탈과립유도 효과도 감소시킴을 확인할 수 있었다. 이는 만병초와 구상나무 추출물이 항알러지 효과가 있음을 제시하는 결과이다.

• BMB Reports
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• 제55권6호
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• pp.275-280
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• 2022

The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.

• 대한한의학회지
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• 제23권3호
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• pp.104-118
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• 2002

Objectives : To study the effects of Kumsooyukkun-jeon on asthma. Methods : Asthma was induced to Balb/c mice with ovalbumin using the method of Hatfield et al. We measured the 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 mucous-secretory cells in the bronchus, morphology of the bronchus, ultramicroscopical appearance of surface of trachea and number of cilia and mucous-secretory cells by scanning electron microscope. Results : 1. Hypertrophy of mucous membrane of trachea and bronchus and bronchioles in the lung, peritracheal, peribronchus and peribronchiolar inflammatory cell infiltration, and mucoid exudate deposition the lumen were observed in control groups but these phenomena were recovered in the Kumsooyukkun-jeon groups. 2. Cellular compartments including neutrophil and eosinophil were increased in the BALF of control groups but these phenomena were recovered in the Kumsooyukkun-jeon groups. 3. Degranulation and decrease of the numbers of mast cells were detected in the trachea of control groups. However, these phenomena were recovered in the Kumsooyukkun-jeon groups. 4. Shedding, decrease of cilia cell and increase of mucous-secretory cells in the surface of the trachea were measured in control groups but these phenomena were recovered in the Kumsooyukkun-jeon groups. Conclusions : It is considered that Kumsooyukkun-jeon has somewhat favorable effects on asthma.

• 한국미생물·생명공학회지
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• 제33권4호
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• pp.322-326
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• 2005

IgE-dependent histamine-releasing factor (HRF) is found extracellularly to regulate the degranulation process of histamine in mast cells and basophils and known to play a predominant role in the pathogenesis of chronic allergic disease. HRF has been also identified in the intracellular region of the cell. Previously, we reported that HRF interacts with the 3rd cytoplasmic domain of the alpha subunit of Na,K-ATPase and inhibits Na,K-ATPase activity. Since it is known that estroaen activates the sarcolemmal Na,K-ATPase, we tested whether estrogen recovers the Na,K-ATPase activity repressed by HRF. In this study, we showed that estrogen activates Na,K-ATPase repressed by HRF. RT-PCR and western blot analysis showed that estrogen doesn't reduce the expression level of HRF in HeLa cell, suggesting that this recovery effects of estrogen probably occur via indirect mechanism on HRF and Na,K-ATPase.

• Environmental Analysis Health and Toxicology
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• 제14권3호
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• pp.95-101
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• 1999

The scavenging effects of two hyaroxycinnamic acids such as caffeic acid and chlorogenic acid on paraquat induced pulmonary toxicity were investigated. The results are summerized as follows: 1. In the 5-lipoxygenase assay, caffeic acid and chlorogenic acid inhibited the enzyme activities whose inhibition concentration (IC$\_$50/) were 4.1 and 9.6 ${\mu}$M respectively. 2. To evaluate the antiinflammatory effects on mediator related to the mechanism of inflammation, ADP-induced platelet aggregation assay and histamine degranulation assay were used. Caffeic acid and chlorogenic acid inhibited on ADP-induced platelet aggregation and histamine release at a concentration dependent manners. 3. Arachidonic acid-induced ear edema were inhibited by administration of caffeic acid and chlorogenic acid. 4. Cytologicad analysis of branchoalveolar lavage fluid (BALF) which was the useful tool for detection of an inflammatory response in the lungs of animals intoxicated with chemicals were used. Alveolar macrophages and neutrophils in BALF, as well as the protein content and the LDH activity in BALF supernatant increased by intoxication of paraquat, but decreased by administration of caffeic acid and chlorogenic acid. Therefore, two hydroxyeinnamic acids tested were the useful candidates for scavenger and antiinflammatory agents on paraquat induced pulmonary toxicity.