• 대한본초학회지
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• 제24권3호
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• pp.29-37
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• 2009

Objectives : The present study was carried out to investigate the effect of Trichosanthis Radix extract (TRE) on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mice by ovalbumin (OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dossiciated with collagenase (1 $\mu$g/ml). Eosinophils were activated by rmIL-3/rmIL-5 co-treatments. The lung cells were treated with TRE, incubated for 48 hr at 37$^{\circ}C$, and analyzed by flow cytometer, ELISA and RT-PCR methods Results : To measure cytotoxicity, mouse lung fibroblast cells (mLFCs) were pretreated with various concentrations of TRE. TRE at 100 $\mu$g/ml, the highest concentration, examined did not have any cytotoxic effects on mLFCs. In FACS analysis, number of granulocyte/lymphocyte, CD3e-/CCR3+, CD3e+/CD69+, CD4+/CD8+ T cells in asthma-induced lung cells were significantly decreased by TRE treatment compared to the control group. But CD4+/CD25+ T cells were not examined significant change in lung cells treated with TRE. In ELISA analysis, production levels of IL-3, IL-5, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by TRE treatment. Conclusions : The present data suggested that Trichosanthis Radix on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Trichosanthis Radix.

• Clinical and Experimental Pediatrics
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• 제54권9호
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• pp.373-379
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• 2011

Purpose: 4-1BB (CD 137) is a costimulatory molecule expressed on activated T-cells. Repression by 4-1BB is thought to attenuate Th2-mediated allergic reactions. The aim of this study was to investigate the effect of 4-1BB on allergic airway inflammation in a murine asthma model. Methods: BALB/c mice were sensitized to and challenged with ovalbumin (OVA). Hu.4-1BB-Fc was administered 1 day before the first OVA sensitization or 1 day after the second OVA sensitization. Following antigen challenge, airway responsiveness to methacholine was assessed and bronchoalveolar lavage (BAL) fluid was analyzed. Total immunoglobulin (Ig) E, OVA-specific IgE, $IgG_1$, and $IgG_{2a}$ levels in sera were measured by enzyme-linked immunosorbent assay. Lung pathology was also evaluated. Results: In mice treated with Hu.4-1BB-Fc before the first OVA sensitization, there was a marked decrease in airway hyperresponsiveness, total cell count, and eosinophil count in the BAL fluid. In addition, Hu.4-1BB-Fc treatment decreased serum OVA-specific $IgG_1$ levels and increased serum $IgG_{2a}$ level significantly compared with the corresponding levels in mice sensitized to and challenged with OVA. Hu.4-1BB-Fc-treated mice also showed suppressed peribronchial and perivascular inflammatory cell infiltration. In contrast, treatment with Hu.4-1BB-Fc 1 day after sensitization had no effect on airway hyperresponsiveness and showed less suppression of inflammation in lung tissue. Conclusion: Administration of Hu.4-1BB-Fc can attenuate airway inflammation and hyperreactivity in a mouse model of allergic airway inflammation. In addition, administration before sensitization may be more effective. These findings suggest that 4-1BB may be a useful therapeutic molecule against asthma.

• 동의생리병리학회지
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• 제24권4호
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• pp.616-623
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• 2010

Three herbal formulas (Bangpungtongsung-san, Ohyaksungi-san, and Ojeok-san) for wind-cold and heat pain symptom were applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses in murine macrophage cell line (RAW 264.7) and ovalbumin (OVA)-immunized mice. This study was carried out in nitric oxide (NO) synthesis in RAW 264.7 cells and cellular proliferation in mouse splenocytes according to three herbal formulas. C57BL/6 mice were immunized intraperitonially with OVA/aluminium ($100\;{\mu}g/200\;{\mu}g$/mouse) on day 1, 8, and 15. Three herbal formulas were administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, lgG1, and total IgM classes) in plasma were measured. Ohyaksungi-san increased NO synthesis in RAW 264.7 cells. Ojeok-san and Ohyaksungi-san significantly enhanced cellular proliferation by LPS and Con A in splenocytes from OVA-immunized mice (p<0.001). Three herbal formulas for wind-cold and heat pain symptom also significantly enhanced plasma OVA-specific IgG, IgG1, and total IgM levels compared with the OVA/Alum group. These results suggested that three herbal formulas for wind-cold and heat pain symptom could be used as stimulator of immune response.

• 대한한의학회지
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• 제32권3호
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• pp.25-34
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• 2011

Objectives: We performed simultaneous determination of five constituents by HPLC in Samul-tang (SMT). Additionally, we investigated the immune-stimulatory potential of SMT on specific cellular and humoral immune responses in ovalbumin (OVA)-immunized mice. Methods: Reverse-phase chromatography using a Gemini C18 column operating at $40^{\circ}C$, and photodiode array (PDA) detection at 190-400 nm, were used for quantification of the five components of SMT. Mobile phase using a gradient flow consisted of two solvent systems. Solvent A was 1.0% (v/v) aqueous acetic acid and solvent B was acetonitrile with 1.0% (v/v) acetic acid. C57BL/6 mice were immunized intraperitoneally with OVA/alum ($100{\mu}g/200{\mu}g$) on days 1, 8, and 15. The extract of SMT (1000 mg/kg) was given to mice orally for 21 days (from day 1 to day 21). At day 22, OVA-, lipopolysaccharide (LPS)- and concanavalin A (Con A)-stimulated splenocyte proliferation and OVA-specific and total antibodies were measured in plasma. Results: Calibration curves were acquired with $r^2$>0.9999, and the relative standard deviation (RSD, %) for intra- and inter-day precision were both less than 3.5%. The recovery was in the range of 95.69-115.12%, with an RSD less than 6.0%. The contents of five components in SMT were 1.08-15.30 mg/g. SMT significantly enhanced Con A-induced splenocyte proliferation in OVA-immunized mice (p<0.01). Also, SMT significantly enhanced OVAspecific IgG, IgG1 and total IgM levels in plasma compared with the OVA-immunized group. Conclusions: The established method will be applied for the quantification of major components and immunestimulating activity in OVA-immunized mouse model of SMT.

• 대한한방소아과학회지
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• 제21권1호
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• pp.41-51
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• 2007

목적 : OVA에 유도된 천식 쥐 모델에 향소산가미방을 투여한 후 $PPAR{\gamma}$의 변화를 조사하여 향소산가미방의 천식 치료 기전을 알아보고자 하였다. 대상 및 방법 : 8주된 암컷 BALB/c 마우스에 첫 날과 14일 후에 20 ${\mu}g$의 OVA를 알루미니윰 하이드록사이드 1mg과 혼합한 후 총 200 ${\mu}g$를 복강 내로 주입하여 감작시켰다. 처음 감작시킨 날로부터 21,22,23일 후에 천식 모델에 사용하는 초음파 분무기를 이용하여 세 번째 감작시켜서 천식 마우스 모델을 만들었다. 천식 마우스 모델을 만드는 기간 중 OVA를 복강 내로 주입한 후 19일째에 24시간의 간격으로 향소산가미방을 7일 동안 경구 투여하여 향소산가미방의 효과를 조사하였다. 기관지폐포세척술은 마지막 감작 후 72시간 후에 실행하고 기관지폐포 세척액의 총 세포수를 측정하였다. $PPAR{\gamma}$의 발현은 천식 마우스 모델의 폐와 향소산가미방을 투여한 마우스 모델의 폐를 적출한 후 Western blotting 방법을 아용하여 측정하였다. 병리 조직학적 검사는 hematoxylin 2 and eosin-Y 염색을 이용하여 조사하였다. 결과 : 정상 군과 비교하여 OVA감작 천식 쥐 모델에서는 72시간 후에 총 세포 수가 증가하였다. 특히 OVA감작 천식 군에서 증가된 호산구의 수가 향소산가미방을 투여 한 쥐 군에서는 유의하게 감소하였다. OVA감작 천식 쥐 모델에서 72시간 후에 정상 군과 비교하여 핵 내에서의 $PPAR{\gamma}$단백질의 발현이 약간 증가하였다. 그러나 향소산가미방을 투여한 쥐 모델에서는 세포질과 핵 내에서 $PPAR{\gamma}$단백질의 발현이 유의하게 증가하였다. 조직학적 검사상 정상 군과 비교하여 OVA 감작된 천식 쥐 모델에서는 폐포, 세기관지, 기도내강 주변에 많은 염증 세포들이 있었다. 그러나 가미향소산 을 투여 한 후에는 염증 세포들이 유의하게 감소하였다. 결론 : 가미향소산은 $PPAR{\gamma}$작용제로서 역할을 하며, 천식에 대한 치료제 또는 예방제를 개발하는 데 후보 물질이 될 것으로 사료된다.

• 동의생리병리학회지
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• 제31권5호
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• pp.277-283
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• 2017

Activated T helper 2 (Th2) immune function is hallmark of various allergic diseases. We investigated the anti-allergic effect of Jagamcho-tang extract(JE) on ovalbumin(OVA)-induced atopic dermatitis mice model and OVA-stimulated splenocytes isolated from the mice. Mice were intraperitoneally injected OVA/alum solution 2 times at interval of 14 days, followed by oral administration of JE for 7 days. After administration, mice were subcutaneously injected with OVA in ear. JE treatment reduced ear swelling and infiltration of inflammatory cells in ear. Serum levels of interleukin(IL)-4 and immunoglobulins, such as total-IgE and OVA-specific IgE, were decreased in JE treated group. Furthermore, JE treatment decreased OVA-induced Th2-associated cytokines like IL-4, IL-5 and IL-13 mRNA levels in splenocytes. In conclusion, JE reduced allergic immune response via IgE production and Th2 response in OVA-sensitized mice, suggesting that JE could be useful prescription for allergic diseases including atopic dermatitis.

• 한국동물학회지
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• 제13권3호
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• pp.68-74
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• 1970

안전방안에서 여포난자가 성숙한다는 것이 몇가지 동물을 이용한 실험에서 밝혀진 바가 있었거니와 여포 난자가 異種의 안전방내에서도 성숙이 가능한 가를 보기 위하여 본 실험을 행하였다. 토끼의 경우 자체의 안전방내에 이식한 여포난자가 다른 동물 즉 흰쥐나 흰생쥐의 안전방에 이식하였을 때보다 낮은 성숙률을 보여 주고 있을 뿐, 흰쥐, 흰생쥐들은 각각 상호간의 안전방에 이식하였을때 同種의 다른 개체의 안정방에서 얻은 결과보다 훨씬 좋은 성숙률을 나타내고 있다. 특히 흰쥐의 여포난자는 숫컷 흰쥐의 안전방에서 24시간 사이에 76%가 제 1및 제 2차 분렬의 증기에 있으며 같은 종류의 수컷 생쥐의 안전방에서 55%가 성숙한 것 보다 훨씬 높은 율을 보여주고 있는 것이다. 안전방의 전방수가 함유단백질의 양이거나 삼투압에 있어서 일반체액이거나 표준배양액의 것들과는 크게 다르며 세포나 조직의 배양액으로는 그다지 적당한 것으로 보이지 않으나 여포난자의 활성획득에는 큰 영향이 없다할 것이다. 특히 異種간의 이식실험에서 얻은 결과가 자체거나 同種간의 이식실험과 거의 같거나 오히려 더 나은 성과를 보여주고 있는데, 는 그 이유에 대해 더 밝혀야 할 문제로 본다. 단지 난자는 성숙분렬을 위한 활성을 갖는데 난자자체의 구성성분 즉 난황물질의 양에 크게 좌우되는 것같으며 또한 난자가 배양될 안전방의 환경요인이 성숙률을 결정하는 데 큰 영향을 주는 것으로 보인다.

• 대한한방소아과학회지
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• 제26권4호
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• pp.10-23
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• 2012

Objectives: Gagamyanggyeoksan (G-YGS) has been used to suppress allergic reaction, however, the cellular target of G-YGS and its mode of action remain unclear. The present study was designed to investigate the effect of extracted G-YGS on the PMA and lonomycin (PI)-induced activation of RBL-2H3. Methods: For this investigation, We examined IL-4, IL-13 mRNA expression by Real-Time PCR, IL-4, IL-13 production by ELISA analysis and manifestations of GATA-1, GATA-2, NF-AT1, NF-AT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting, OVA-specific IgE, IL-4, IL-13 by mouse be sensitive to OVA. Results: Here we showed that treatment of RBL-2H3 mast cells with G-YGS, suppressed PI-induced production of Th2 cytokines including IL-4 and IL-13 in a dose dependent manner. The mRNA expression of IL-4 were completely abolished by G-YGS at the concentration of $100{\mu}g/ml$. Data from a stable cell lines consistently expressing IL-4. And the mRNA expression of IL-13 were abolished by G-YGS at the $200{\mu}g/ml$. But there is no difference between the $50{\mu}g/ml$, the $100{\mu}g/ml$ and the comparison. Results from the western blot analysis of transcription factors involving IL-4 and IL-13 expression indicated that it prominently decreased the expression of mast cell specific transcricption factors including GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65 but not c-Fos. And G-YGS suppressed IgE, IL-4, IL-13 in mouse be sensitive to OVA. Conclusions We suggested the anti-allergic activities of G-YGS might be mediated by down-regulation of Th2 cytokines such as IL-4 and IL-13 through the regulation of transcription factors as GATA-1, GATA-2, NF-AT2, c-Jun, NF-${\kappa}B$ p65.

• Tuberculosis and Respiratory Diseases
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• 제84권1호
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• pp.55-66
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• 2021

Background: Particulate matter 10 (PM₁₀; airborne particles <10 ㎛) inhalation has been demonstrated to induce airway and lung diseases. In this study, we investigate the effects of PM₁₀ inhalation on RNA expression in lung tissues using a murine model. Methods: Female BALB/c mice were affected with PM₁₀, ovalbumin (OVA), or both OVA and PM₁₀. PM₁₀ was administered intranasally while OVA was both intraperitoneally injected and intranasally administered. Treatments occurred 4 times over a 2-week period. Two days after the final challenges, mice were sacrificed. Full RNA sequencing using lung homogenates was conducted. Results: While PM₁₀ did not induce cell proliferation in bronchoalveolar fluid or lead to airway hyper-responsiveness, it did cause airway inflammation and lung fibrosis. Levels of interleukin 1β, tumor necrosis factor-α, and transforming growth factor-β in lung homogenates were significantly elevated in the PM₁₀-treated group, compared to the control group. The PM₁₀ group also showed increased RNA expression of Rn45a, Snord22, Atp6v0c-ps2, Snora28, Snord15b, Snora70, and Mmp12. Generally, genes associated with RNA splicing, DNA repair, the inflammatory response, the immune response, cell death, and apoptotic processes were highly expressed in the PM₁₀-treated group. The OVA/PM₁₀ treatment did not produce greater effects than OVA alone. However, the OVA/PM₁₀-treated group did show increased RNA expression of Clca1, Snord22, Retnla, Prg2, Tff2, Atp6v0c-ps2, and Fcgbp when compared to the control groups. These genes are associated with RNA splicing, DNA repair, the inflammatory response, and the immune response. Conclusion: Inhalation of PM₁₀ extensively altered RNA expression while also inducing cellular inflammation, fibrosis, and increased inflammatory cytokines in this murine mouse model.

• The Korean Journal of Pain
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• 제35권2호
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• pp.140-151
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• 2022

Background: Essential oils are of great interest for their analgesic and anti-inflammatory properties. We aimed to study the content of the essential oil of the Origanum vulgare of the Armenian highlands (OVA) in different periods of vegetation and to investigate its antinociceptive and anti-inflammatory effects in mice (in vivo) and cytotoxic action in cultured cells (in vitro). OVA essential oil was extracted from fresh plant material by hydro-distillation. Methods: For OVA essential oil contents determination the gas chromatography-mass spectrometry method was used. Formalin and hot plate tests and analysis of cell viability using the methyl-thiazolyl-tetrazolium (MTT) assay were used. Results: The maximal content of β-caryophyllene and β-caryophyllene oxide in OVA essential oil was revealed in the period of blossoming (8.18% and 13.36%, correspondently). In the formalin test, 4% OVA essential oil solution (3.5 mg/mouse) exerts significant antinociceptive and anti-inflammatory effects (P = 0.003). MTT assay shows approximately 60% cytotoxicity in HeLa and Vero cells for 2.0 µL/mL OVA essential oil in media. Conclusions: The wild oregano herb of Armenian highlands, harvested in the blossoming period, may be considered as a valuable source for developing pain-relieving preparations.