Objective : Allergic asthma is a chronic airway disease that affects millions of people in the developed world. The disease is characterized by concurring airway inflammation, Th2 cytokine production, increased mucus secretion, airway hyperresponsiveness (AHR) to inhaled antigen, and pulmonary fibro...
Objective : Allergic asthma is a chronic airway disease that affects millions of people in the developed world. The disease is characterized by concurring airway inflammation, Th2 cytokine production, increased mucus secretion, airway hyperresponsiveness (AHR) to inhaled antigen, and pulmonary fibrosis. To investigate the therapeutic and anti-asthmatic effects of Drynariae Rhizoma (DR), we examined the influence of DR on the development of pulmonary eosinophilic inflammation and airway hyperresponsiveness in a mouse model of allergic asthma. Methods : In this study, BALB/c mice were systemically sensitized to ovalbumin (OVA) followed intratracheally, intraperitoneally, and by aerosol allergen challenges. We investigated the effect of DR on airway hyperresponsiveness, pulmonary eosinophilic infiltration, various immune cell phenotypes, Th2 cytokine production and OVA specific IgE production in a mouse model of asthma. Results : In asthmatic mice, we found that DR.treated groups had suppressed eosinophil infiltration, allergic airway inflammation and AHR by suppressing the production of IL-5, IL-13 and OVA specific IgE. Conclusions : Our data suggest that the therapeutic mechanism by which DR effectively treats asthma is based on reductions of Th2 cytokines (IL-5), eotaxin, OVA-specific IgE production and eosinophil infiltration.
Objective : Allergic asthma is a chronic airway disease that affects millions of people in the developed world. The disease is characterized by concurring airway inflammation, Th2 cytokine production, increased mucus secretion, airway hyperresponsiveness (AHR) to inhaled antigen, and pulmonary fibrosis. To investigate the therapeutic and anti-asthmatic effects of Drynariae Rhizoma (DR), we examined the influence of DR on the development of pulmonary eosinophilic inflammation and airway hyperresponsiveness in a mouse model of allergic asthma. Methods : In this study, BALB/c mice were systemically sensitized to ovalbumin (OVA) followed intratracheally, intraperitoneally, and by aerosol allergen challenges. We investigated the effect of DR on airway hyperresponsiveness, pulmonary eosinophilic infiltration, various immune cell phenotypes, Th2 cytokine production and OVA specific IgE production in a mouse model of asthma. Results : In asthmatic mice, we found that DR.treated groups had suppressed eosinophil infiltration, allergic airway inflammation and AHR by suppressing the production of IL-5, IL-13 and OVA specific IgE. Conclusions : Our data suggest that the therapeutic mechanism by which DR effectively treats asthma is based on reductions of Th2 cytokines (IL-5), eotaxin, OVA-specific IgE production and eosinophil infiltration.
* AI 자동 식별 결과로 적합하지 않은 문장이 있을 수 있으니, 이용에 유의하시기 바랍니다.
문제 정의
Although the precise mechanism of action needs further investigation, this is the first report that DR significantly inhibited airway inflammation and hyperresponsiveness in a mouse model of allergic asthma. Therefore, we propose that DR can be used for the treatment of pathologic inflammatory airway disorders such as allergic asthma.
The aim of this study was to evaluate the control activity of DR extract on Th1- and Th2-type cytokines, the development of allergic airway inflammation and airway hyperresponsiveness. Therefore, we decided to investigate antiinflammatory and antiasthmatic effects of DR extract in a mouse model of allergic asthma.
제안 방법
At the end of the experiment,, lung was fixed and histologic analysis was performed. Lung sections of normal, asthmatic mice treated with vehicle (control), DR (200 mg/kg), and DR (50 mg/kg).
) for flow cytometric analysis were purchased from Becton Dickinson (BD) PharMingen (SanDiego,CA). Cells from lung tissues and BALF were stained with the indicated antibodies in staining buffer (PBS containing 1% FBS and 0.01% NaN3)for 10 min on ice, and analyzed by two color flow cytometry on a FACSCalibur using Cell Quest software(BD Biosciences, MountainView, CA) for data expression.
Penh is equal to Pause × PEF/PIF, where Pause = (Te-Tr)/Tr (PIF, peak inspiratory flow； PEF： peak expiratory flow； Te, expiratory time；Tr, relaxation time). In this experiment mice were aerosolized with OVA for 30 min/day, 3 days/week for 5 weeks. At 24 hours after the final inhalation, mice were given aerosolized normal saline, followed by 3.
DR (50 and 200 mg/kg) were orally administered 3 times a week for the last 5 weeks. One day after the last OVA exposures (2％ OVA inhalation), airway hyperresponsiveness was determined and samples (bronchoalveolar lavage fluid, lung cells, and serum) were collected for further molecular analyses.
Korea) in PBS. Seven days after the second sensitization intratracheally injected with 250㎍ of OVA (on day 21) on the back of the tongue, mice were exposed to aerosolized OVA for 30 min/day, 3 days/week for 5 weeks (at a flow rate of 250 L/min, 1％ OVA in normal saline for first 4 weeks and 2％ OVA in normal saline for last 1 week). DR (50 and 200 mg/kg) were orally administered 3 times a week for the last 5 weeks.
0 statistic software). The difference between the normal group and the control group (OVA+vehicle) was clearly distinguished, and for this reason, statistical significance between the normal group and the control group was not shown in the figures and tables to put an emphasis on the statistical differences between the experimental groups and the control group. Results (presented as mean ± standard error of mean) were considered statistically significant if P values were <0.
To determine whether DR influenced cytokine and chemokine secretion in the BALF, the levels of IL-4, IL-5, and IL-13 in this fluid were measured using ELISA after the final challenge.
To investigate the level of mucus expression in the airway, PAS-positive and PAS-negative epithelial cells in individual bronchioles were analyzed. The degrees of goblet cell hyperplasia and mucus hyperproduction were evaluated by means of PAS staining and quantification of PAS-stained cells.
As described in Materials and Methods, lung and BALF was harvested 24 hrs after the last OVA challenge. Total inflammatory cell numbers in lung and BALF were counted, and cell classification was performed on a minimum of 200 cells to classify lymphocytes. Results are expressed as mean±S.
데이터처리
Data were analyzed by one-way analysis of variance (ANOVA) or unpaired Student's t-test followed by Dunnett's multiple comparison test (SPSS version 14.0 statistic software).
Differences of Penh value between groups were evaluated using a unpaired Student’s t-test.
Statistical significance between control and treatment groups was assessed by ANOVA or unpaired Student's t-test followed by Dunnett's multiple comparison test (*p<0.05, **p<0.01, ***p<0.001).
성능/효과
As previously described in results, effects of DR on leukocyte subsets in lungs and BALF were marked with change in numbers of CD3+ T cells, CD4+ helper T cells, Gr-1+/CD11b+granulocytes,CD3-/CCR3+ eosinophils, CD3+/CCR3+ Th2 cells, CD3 +/CD69+ early activated T cells, B220+/CD23+ B cells in a mouse model of asthma compared to control group (Table 1), and the deficits in CD3-/CCR3+ eosinophils were accompanied by concurrent decreases eosinophils in BALF cytospin (Fig. 3d). DR also inhibit B cell– dependent production of OVA specific IgE in serum (Fig.
Moreover, eosinophils are one of the cell types known to express Gr-1, therefore eosinophil populations may constitute a substantial portion of the CD11b+Gr-1+ populations. Our results(Table 1) showed that Gr-1+ cells was increased with OVA challenge but significantly decreased with DR treated mice in BALF and lung.
To evaluate effect of DR on immune cell subtypes flow cytometric analysis was accomplished. The numbers of CD3, CD4, CD8, CCR3, CD69, B220, CD23, CD11b, Gr-1 positive cells in the lungs of OVA-challenged mice were increased compared to the saline treated group, and generally, each values from DR-treated mice were significantly lower than those of OVA-challenged mice (Table 1). Formoterol administration resulted significant reduction in T cell subtypes similarly.
후속연구
It would be valuable to find the natural products that have specific inhibitory effects on airway inflammation and hyperresponsiveness and Th2 cytokines production – in view of both basic and clinical sciences – and the result from this study suggest a possibility of using DR as new therapeutic material for respiratory diseases including allergic asthma, although further studies are required.
참고문헌 (27)
Kay AB. Asthma and inflammation. J. Allergy. Clin. Immunol. 1991 ; 87 : 893-910.
Bousquet J, Jeffery PK, Busse WW, Johnson M, Vignola AM. Asthma. From bronchoconstriction to airways inflammation and remodeling. Am. J. Respir. Crit. Care Med. 2000 ; 161 : 1720-45.
Lopez AF, Sanderson CJ, Gamble JR, Campbell HD, Young IG, Vadas MA. Recombinant human interleukin 5 is a selective activator of human eosinophil function. J. Exp. Med. 1988 ; 167 : 219-24.
Sanderson CJ, Warren DJ, Strath M. Identification of a liymphokine that stimulates eosinophil differentiation in vitro. Its relationship to interleukin-3, and functional properties of eosinophils produced in cultures. J. Exp. Med. 1985 ; 162 : 60.
Nakajima H, Hirose K. Role of IL-23 and Th17 Cells in Airway Inflammation in Asthma. Immune Netw. 2010 ; 10 : 1-4.
Shi XC. Chinese-English Terminology of Traditional Chinese Medicine. Hunan Science Publishing Co., China. 1983 : 71-98.
Park SM, Lee JH, Lee KC, Yang HW, Lee BC, Lee JH, Jeong JG, Seo BI, Kang YS, Kim SS, Hwang DR, Kim DH, Shin MK, Song HJ. Effect of Drynariae Rhizoma on Cultured Mouse Fibroblast Injured by Streptomycin. The Korea Journal of Herbology. 2003 ; 18(1) : 7-13.
Liang YH, Ye M, Zhang LZ, Li HF, Han J, Wang BR, Guo DA. Two new phenolic acids from Drynariae Rhizoma. Acta Pharm. Sin. 2010 ; 45 : 874-878.
Liang YH, Wei W, Yu SW, Ye M, He XH, Gong NB, Lu Y, Ikhlas AK, Guo DA. A new chiratane type triterpenoid from the rhizomes of Drynaria fortunei. Fitoterapia. 2010 ; 81 : 988-991.
Liang YH, Ye M, Han J, Wang BR, Guo DA. Lignans and flavonoids from rhizome of Drynaria fortunei. Chin. Trad. Herb. Drugs. 2011 ; 42 : 25-30.
Kim SH, Kim BK, Lee YC. Antiasthmatic effects of hesperidin, a potential Th2 cytokine antagonist, in a mouse model of allergic asthma. Mediators Inflamm. 2011 ; 2011 : 485402.
Bharadwaj A, Agrawal DK. Immunomodulation in asthma : a distant dream or a close reality? Int. Immunopharmacol. 2004 ; 4 : 495-511.
Lee NA, Gelfand EW, Lee JJ. Pulmonary T cells and eosinophils : coconspirators or independent triggers of allergic respiratory pathology? J. Allergy Clin. Immunol. 2001 ; 107 : 945-57.
Yamaguchi Y, Suda T, Suda J, Eguchi M, Miura Y, Harada N, et al. Purified interleukin 5 supports the terminal differentiation and proliferation of murine eosinophilic precursors. J. Exp. Med. 1988 ; 167 : 43-56.
Lopez AF, Sanderson CJ, Gamble JR, Campbell HD, Young IG, Vadas MA. Recombinant human interleukin 5 is a selective activator of human eosinophil function. J. Exp. Med. 1988 ; 167 : 219-24.
Rothenberg ME, Hogan SP. The eosinophil. Ann. Rev. Immunol. 2006 ; 24 : 147-74.
Gleich GJ, Frigas E, Loegering DA, Wassom DL, Steinmuller D. Cytotoxic properties of the eosinophil major basic protein. J. Immunol. 1979 ; 123 : 2925-2927.
Humbles AA, Conroy DM, Marleau S, Rankin SM, Palframan RT, Proudfoot AE, Wells TN, Li D, Jeffery PK, Griffiths-Johnson DA, et al. Kinetics of eotaxin generation and its relationship to eosinophil accumulation in allergic airways diseas e : analysis in a guinea pig model in vivo. J. Exp. Med. 1997 ; 186 : 601.
Kitamura M, Nakajima T, Imai T, Harada S, Combadiere C, Tiffany HL, Murphy PM, Yoshie O. Molecular cloning of human eotaxin, an eosinophil-selective CC chemokine, and identification of a specific eosinophil eotaxin receptor, CC chemokine receptor 3. The journal of biological chemistry. 1996 ; 271 : 7725-7730.
Umland SP, Wan Y, Shortall J, Shah H, Jakway J, Garlisi CG, Tian F, EganRW, Billah MM. Receptor reserve analysis of the human CCR3 receptor in eosinophils and CCR3-transfected cells. Journal of leukocyte biology. 2000 ; 67(3) : 441-447.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.