• The Journal of Internal Korean Medicine
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• v.28 no.4
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• pp.681-693
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• 2007

Background and Objective : Atractylodes japonica (AJ) is a commonly-used herbal medicine in Asian countries such as Korea, China and Japan. The present study was designated to evaluate the direct effects of AJ on helper T cell activities and on Th1/Th2 lineage development in vitro. Materials and Methods : Spleen cells from 8-week BALB/c mice were cultured in CR extracts containing medium without activation for 24 hours and with activation for 48 hours. CD4+ T cells were isolated and analyzed for mRNA expression levels of INF-$\gamma$, IL-4, T-bet and GATA-3 by RT-PCR and secretion cytokines levels of INF-$\gamma$, IL-2, IL-4, IL-5 and IL-10 by ELISA. Results : The results demonstrated that AJ had no mitogenic effects on unstimulated CD4+ T cells, but augmented CD4+T-cell proliferation upon activation with anti-CD3/anti-CD28 antibodies in a dose-dependent manner. AJ treatment significantly increased CD4+ T cell population and IFN-$\gamma$ expression was significantly enhanced, while IL-4 expression significantly decreased. In addition, in vitro Th1/Th2 polarization experiments revealed that AJ enhanced IFN-$\gamma$ secretion in Th1 cells, but reduced the IL-4 in Th2 cells in dose-dependent manner. Conclusion : These results suggest that AJ treatment could be a desirable alternative therapy for the prevention or correction of Th2 dominant pathological disorders, such as allergy and asthma.

• The Journal of Internal Korean Medicine
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• v.30 no.3
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• pp.451-464
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• 2009

Background and Objective : Cyperus rotundus L. (CR) is a commonly used herbal medicine in Asian countries such as Korea, China and Japan. The present study was designated to evaluate the direct effects of CR on helper T cell activities and on Th1/Th2 lineage development in vitro. Materials and Methods : Spleen cells from 8 week BALB/c mice were cultured in CR extracts containing medium without activation for 24 hours and with activation for 48 hours. CD4+ T cells were isolated and analyzed for mRNA expression levels of INF-$\gamma$, IL-4, T-bet and GATA-3 by RT-PCR and secretion cytokines levels of INF-$\gamma$, IL-4, IL-5 and IL-10 by ELISA. Results : The results demonstrated that CR had no mitogenic effects on unstimulated CD4+ T cells, but augmented CD4+ T-cell proliferation upon activation with anti-CD3/anti-CD28 antibodies in a dose-dependent manner. CR treatment significantly increased CD4+ T cell population and the IFN-$\gamma$ expression was significantly enhanced, while IL-4 expression was significantly decreased. In addition, in vitro Th1/Th2 polarization experiments revealed that CR enhanced IFN-$\gamma$ secretion in Th1 cells, but reduced the IL-4 in Th2 cells in a dose-dependent manner. Conclusion : These results suggest that CR treatment could be a desirable alternative therapy for the prevention or correction of Th2 dominant pathological disorders, such as allergy and asthma.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.5
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• pp.1397-1403
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• 2004

By recently study, GM (Gamipaemo-tang) treatment have worked well on the allergic asthma. The purpose of this study was effect of GM extract on helper T cell, major regulator of immune system. Splenic cells from 8-week BALB/c mice were cultured in GM containing media without activation for 48 hours. The MTS assay and flow cytometry study revealed that lymphocyte treated with GM were not effective on CD4+ T cells. Subsequently CD4+ T cells were isolated and cultured in GM containing media. Either GM were not effective on CD4+ T cell without APCs. By FACS scan analysis, the expression of INF-γ, IL-4 were down-regulated in the condition skewed Th1 and Th2 cells respectively, Using ELISA analysis, the expression of INF-γ is up-regulated and IL-4 is down-regulated in the condition skewed Th1, Th2 cells respectively. With RT-PCR analysis, the expression of mRNA for INF-γ is down-regulated and IL-4 is down-regulated in the condition skewed Th1 and Th2 cells respectively. The result suggests that GM inhibited the differetiation of Th2 cells significantly and indicates GM could enhance anti-allergic immune system.

• The Korea Journal of Herbology
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• v.24 no.1
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• pp.79-88
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• 2009

Objectives : It has been recently shown that Piperis Longi Fructus (PLF) is involved in the reduction of eosinophil recruitment and production of Th2 cytokines in vivo. However, the main therapeutic mechanisms of PLF remains a matter of considerable debate. To investigate the therapeutic mechanisms of PLF, we examined the influence of PLF on regulatory T cells number, IgE, histamine production in vivo and Th1/Th2 cytokine balance in vitro. Methods : All mice were immunized on two different days (21 days and 7 days before inhalational exposure) by i.p. injections of 0.2 $m\ell$ alum-precipitated Ag containing 100 ${\mu}g$ of OVA bound to 4 mg of aluminum hydroxide in PBS. Seven days after the second sensitization, mice were exposed to aerosolized ovalbumin for 30 min/day on 3 days/week for 12 weeks(at a flow rate of 250 L/min, 2.5％ ovalbumin in normal saline) and PLF (150 mg/kg) were orally administered 3 times a week for 8 weeks. Splenocytes from C57BL/6 mice at 8 weeks of age were stimulated with anti-CD3 (1 mg/ml) plus anti-CD28 (1 mg/ml) antibody for 48hrs. IL-4 and IFN-$\gamma$ in the culture supernatants were measured by ELISA Results : The suppressive effects of PLF on asthma model were demonstrated by the increase the number of regulatory T cells and by reducing IgE, histamine production in vivo and modulation of Th1/Th2 cytokine balance. Conclusions : These results indicate that PLF has a deep inhibitory effects on asthma model mice by increase the number of regulatory T cells, and by reducing IgE, histamine production.

• Tuberculosis and Respiratory Diseases
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• v.45 no.1
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• pp.90-98
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• 1998

Background : The CD4+ T-helper cells comprise functionally distinct subsets of Th1 and Th2 cells that are distinguished on the basis of differential cytokines production Th1 cells secrete interferon-$\gamma$, lymphotoxin, interleukin-2. Th2 cells produce interleukin-4, interleukin-5, interleukin-10. A previous study shown that Th2 cells and their cytokines increased in patients with atopic asthma. We compared cytokines(IL-4, IFN-$\gamma$) activity and subpopulation of T-lymphocytes in peripheral blood from atopic asthmatics versus non-asthmatics. Method: Fifteen patients with atopic asthma(nine men, six women), twelve patients with chronic bronchitis(six men, six women), five healthy persons(three men, two women) were studied. Activity of IL-4, IFN-$\gamma$ and T-cell subpopulation in peripheral blood were estimated. Results: Patients had a median age of 55yr. The mean activity of IL-4 of asthmatics was significantly increased(control $0.75{\pm}1.1pmol/L$, atopic asthmatics $3.50{\pm}0.75pmol/L$, chronic bronchitis $2.01{\pm}1.2pmol/L$), but IFN-$\gamma$ was not significantly increased. In the T lymphocyte sunsets the percent of CD62L+ T-lymphoeytes of asthmatics was not significantly increased (control $16.7{\pm}16.4%$, atopic asthmatics $24.8{\pm}23.6%$, chronic bronchitis $17.0{\pm}16.9%$). Conclusion: In this study elevated production of IL-4 was observed in atopic asthmatics. CD62L+T-lymphoeytes was not increased in atopic asthma.

• Journal of Society of Preventive Korean Medicine
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• v.6 no.1
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• pp.140-155
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• 2002

Background: SBP(桑白皮)is an herbal medicine which has been used in oriental medicine as a traditional therapeutic agent of bronchial asthma. Objective: This study was performed to investigate the effect of SBP on the anti-hypersensitivity and immune response in the murine of type I hypersensitivity induced by the experiment. Materials and Methods: Laboratory rats were primary sensitized with OA(ovalbumin); on day 1, rats of a Control group and Sample group (SBP group) were systemically immunized by subcutaneous injection of 1 mg OA and 300mg of Al(OH)3 in a total volume of 2ml saline. The rats of the sample group were orally administered with an SBP water extract for 14 days after primary immunization. On day 14 after the systemic immunization, rats received local immunization by inhaling 0.9% saline aerosol containing 2%(wt/vol) OA. A day after local immunization, BAL fluid and serum were collected from the rats. Total cell, lymphocyte, CD4+ T cell, CD8+ T cell, CD4+/CD8+ ratio in the BALF, and IgE level in serum were measured and evaluated. Results: SBP showed a suppressive effect on the immune response in the rats. 1. Total cells in the BALF decreased in the SBP treated group in comparision to the control group, but statistic differences were not observed. 2. Total lymphocytes in the BALF were statistically decreased in SBP treated group in comparision to the control group. 3. CD4+ T cells in the BALF were statistically decreased in SBP treated group in comparision to the control group. 4. CD8+ T cells in the BALF were not statistically different in SBP treated group and the control group. 5. The ratio of CD4+/CD8+ in the BALF was statistically decreased in SBP treated group in comparision to the control group. 6. The IgE level in serum decreased in the SBP treated group in comparision to the control group, but statistic differences were not observed.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.185-200
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• 2005

Objective : The aim of this study was to investigate the asthma-suppressive and immune-regulatory effect of AHCR-HA(ASARI HERBA CUM RADICE Herbal-acupuncture) at Pyesu(BLl3) on OVA(ovalbumin)-induced asthma in mice. Methods : C57BL/6 mice out of all the experimental groups, except the Normal group and the AHCR-HA group, were sensitized and challenged with OVA The mice in the AHCR-HA group and the OVA-AHCR-HA group were treated with AHCR-HA(1%) at Pyesu(BL13). The mice in the OVA-Saline group were injected with saline at Pyesu(BL13). The mice in the OVA-Needle-Prick group were treated with a single prick with an injection needle at Pyesu(BL13). AHCR-HA saline injection and needle prick were administered for 8 weeks, three times a week. Result : 1. The populations of granulocytes, CD3e-/CCR3+ cells, CD69+/CD3e+ cells, CD4+ cells and CD23+/B220+ cells in the OVA-induced asthmatic mouse lungs decreased significantly by AHCR-HA. 2. The lung weight, total cells in lung, total leukocytes in BALF, eosinophils in BALF, collagen accumulation in the lung sections of the OVA-AHCR-HA group decreased significantly. 3.The concentrations of IL-4, IL-5, IL-13, IgE in BALF and serum of the OVA-AHCR-HA group decreased significantly. 4. The numbers of Gr-1+/CD11b+, CCR3+, CD3e+, CD19+, CD3e+/CD69+cells in the OVA-AHCR-HA group decreased significantly. 5. The mRAN expressions of $TNF-{\alpha}$, IL-5, IL-4 and IL-13 in lung of the OVA-AHCR-HA group decreased significantly. 6. The AHCR-HA group didn't show any considerable difference from the Normal group. The OVA-saline group and the OVA-Needle prick group showed suppressive effects on OVA-induced asthma however they were not statistically significant. Conclusion : These results suggest that AHCR-HA at Pyesu(BL13) is considered to be effective in treating asthma and to be put to practical use in the future asthma clinic.

• Korean Journal of Acupuncture
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• v.21 no.1
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• pp.61-77
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• 2004

Objectives and methods : The aim of this study was to investigate the effect of AHCR(ASARI HERBA CUM RADICE) herbal acupuncture at St36 on ovalbumin-induced asthma in mice. C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks(once a week). Two experimental groups were treated with different concentrations(1%, 0.1%) of AHCR-HAS at Chok-samni (St36) for the later 8 weeks (3times/week). Results 1. Lung weight of the mice group treated with AHCR-HA decreased significantly compared with that of control group. 2. Total Leukocytes in BALF of the mice group treated wtih AHCR-HA decreased significantly compared with those of control group. 3. The number of Eosinophils in BALF of the mice group treated wtih AHCR-HA decreased significantly compared with that of control group. 4. The number of $Gr-1^+/CD11b^+,\;CCR3^+,\;CD4^+,\;CD3e^+/CD69^+,\;IgE^+/B220^+$ cells in the lungs of the mice group treated with AHCR-HA decreased significantly compared with that of control group. 5. The number of $CD8^+$ cells in the lungs of the mice group treated with AHCR-HA didn't show significant difference compared with that of control group. 6. The concentration of IgE, IL-13, IL-4 in serum of the mice group treated with AHCR-HA decreased significantly compared with that of control group. Conclusion We conclude that AHCR-HA is effective on OVA-induced asthma in C57BL/6 mouse.

• Tuberculosis and Respiratory Diseases
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• v.44 no.4
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• pp.822-835
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• 1997

Background : There have been many in vitro evidences that interleukin-4(IL-4) might be the most important cytokine inducing IgE synthesis from B-cells, and interferon-gamma(IFN-$\gamma$) might be a main cytokine antagonizing IL-4-mediated IgE synthesis. Recently some reports demonstrated that IFN-$\gamma$ might be used as a new therapeutic modality in some allergic diseases with high serum IgE level, such as atopic dermatitis or bronchial asthma. To evaluate the in vivo effect of IFN-$\gamma$ in bronchial asthma we tried a clinical study. Methods : Fifty bronchial asthmatics(serum IgE level over 200 IU/ml) who did not respond to inhaled or systemic corticosteroid treatment, and 17 healthy nonsmoking volunteers were included in this study. The CD 23 expressions of peripheral B-cells, the IL-4 activities of peripheral T-cells, the serum soluble CD23(sCD23) levels, and the superoxide anion(${O_2}^-$) generations by peripheral PMN were compared between bronchial asthmatics and normal subjects. The IL-4 activities of peripheral T-cells were analyzed by T-cell supernatant (T-sup)-induced CD23 expression from tonsil B-cells. In bronchial asthmatics the serum IgE levels and histamine $PC_{20}$ in addition to the above parameters were also compared before and after IFN-$\gamma$ treatment. IFN-$\gamma$ was administered subcutaneously with a weekly dose of 30,000 IU per kilogram of body weight for 4 weeks. Results : The ${O_2}^-$ generations by peripheral PMNs in bronchial asthmatics were higher than normal subjects($8.23{\pm}0.94$ vs $5.00{\pm}0.68\;nmol/1{\times}10^6$ cells, P<0.05), and significantly decreased after IFN-$\gamma$ treatment compared to initial values($3.69{\pm}0.88$ vs $8.61{\pm}1.53\;nmol/1{\times}10^6$ cells, P<0.05). CD23 expression of peripheral B-cells in bronchial asthmatics was higher than normal subjects($47.47{\pm}2.96%$, vs $31.62{\pm}1.92%$, P<0.05), but showed no significant change after IFN-$\gamma$ treatment. The serum sCD23 levels in bronchial asthmatics were slightly higher than normal subjects($191.04{\pm}23.3\;U/ml$ vs $162.85{\pm}4.85\;U/ml$), and 11(64.7%) of 17 patients showed a decreasing pattern in their serum sCD23 levels after IFN-$\gamma$ treatment. However the means of serum sCD23 levels were not different before and after IFN-$\gamma$ treatment. The IL-4 activities of peripheral T-cells in bronchial asthmatics were slightly higher than normal subjects($22.48{\pm}6.81%$ vs $18.90{\pm}2.43%$), and slightly increased after IFN-$\gamma$ treatment($27.90{\pm}2.56%$). Nine(60%) of 15 patients showed a decreasing pattern in their serum IgE levels after IFN-$\gamma$ treatment. And the levels of serum IgE were significantly decreased after IFN-$\gamma$ treatment compared to initial values ($658.67{\pm}120.84\;IU/ml$ vs $1394.32{\pm}314.42\;IU/ml$, P<0.05). Ten(83.3%) of 12 patients showed an improving pattern in bronchial hyperresponsiveness after IFN-$\gamma$ treatment, and the means of histamine $PC_{20}$ were significantly increased after IFN-$\gamma$ treatment compared to initial values ($1.22{\pm}0.29mg/ml$ vs $0.69{\pm}0.17mg/ml$, P<0.05). Conclusion : Our results suggest that IFN-$\gamma$ may be useful as well as safety in the treatment of bronchial asthmatics with high serum IgE level and that in vivo effects of IFN-$\gamma$ may be different from its in vitro effects on the regulations of IgE synthesis or the respiratory burst of PMN.

• Tuberculosis and Respiratory Diseases
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• v.46 no.1
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• pp.44-52
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• 1999

Background : Airway infiltration by inflammatory cells, particularly of eosinophils, is one of the characteristic features of asthma. Several mechanisms for the recruitment of eosinophil is focused on the CD4+ T lymphocyte for the preferential production of Th2-c1erived cytokines. Interleukin-10(IL-10) is identified cytokine with potent antiinflammatory activity. This molecule has been shown to inhibit the release of cytokine from inflammatory cells including Th2 cell, and also to inhibit eosinophil survival. We therefore attempted to determine whether decreased synthesis of IL-10 in the lung of bronchial asthma may contribute to inflammation that is characteristics of this dease. Method: Subjects were patients with bronchial asthma(n=23) and normal controls(n=11). IL-10 produced from peripheral mononuclear cell(PBMC) and in bronchoalveolar lavage(BAL) fluid was measured by ELISA method. Degree of bronchial inflammation was assessed by total cell counts and eosinophil percents in BAL fluid, eosinophil infiltration on bronchial biopsy tissue and $PC_{20}$ for methacholine. Results: The IL-10 level produced by PBMC and in BAL fluid from patient with bronchial asthma were not different with normal controls(respectively, $901.6\pm220.4$ pg/ml, $810.9\pm290.8$ pg/ml for PBMC, $24.5\pm9.5$ pg/mL $30.5\pm13.5$ pg/ml for BAL fluid p>0.05). There were significant negative correlation between IL-10 in BAL fluid and eosinophil percents in BAL fluid or degree of eosinophil infiltration in bronchial biopsy (respectively r=-0.522, r=-0.4486 p<0.05). However there was no difference of IL-10 level according to $PC_{20}$ for methacholine. There were no correlation between IL-10 production by PBMC and peripheral blood eosinophil counts or serum eosinophilic cationic protein levels(respectively r=0.1146, r=0.0769 p>0.05). Conclusion: These observation suggest that IL-10 may participate but not acts the crucial role in regulation of the airway inflammation in bronchial asthma.