1. Suppression of Th2 Immune Responses by Mekabu Fucoidan from Undaria Pinnatifida Sporophylls 김윤지 오지현 최애숙 문정은 이나람애

2. Contents Abstract Introduction Methods Results Discussion

4. What is Fucoidan? Fucoidan is a sulfated polysaccharide found mainly in various species of brown algae and brown seaweed. Fucoidan appears to have anti-tumor, anti-cancer and neuroprotective action. Fucoidan acts to block cancer cells from developing and growing. Fucoidan helps to maintain healthy upper respiratory mucous membranes which when weakened may increase the incidence of Bronchitis, Asthma, Chronic Obstructive Pulmonary Disease and other pulmonary diseases. Fucoidan and Its Benefits

5. Abstract the pulmonary inflammation was relieved by mekabu fucoidan, which also downregulated Th2- dominated responses. Mekabu fucoidan modulates Th2 responses and might be useful for treating allergic inflammation. The production of Th2 cytokines was suppressed, and the amount of IFN-γ was not increased in the mice treated with mekabu fucoidan. Anti-OVA IgE and IgE levels in serum determined after challenge with aerosolized OVA at the end of the experiment were lower in the treated than in the control mice. Mekabu fucoidan was injected intraperitoneally into BALF/c mice for 4 days, and then the mice were sensitized with 50μg/mouse of OVA plus alum (1mg/mouse) 1 and 8 days later. A study was examined the effects of the fucoidan of mekabu on the type 2 T-helper(Th2) response in bronchoalveolar lavage fluid(BALF) after ov-albumin(OVA) aerosol challenge.

6. Introduction Some reports indicate that fucoidan from brown algae exerts antitumor, antibacterial and antiviral activites via several mechanisms in a murine model. Th2 cytokines are important as the allergic inflammatory response leads to airway hyperimmune responsiveness. In other wards, the airway inflammation is due to the production of Th2 cytokines such as IL-4, IL-5 and IL-13 by activated T lymphocytes.

7. In Previous Study It was examined the activity of fucoidan from brown edible algae against L-1210 and P-388 leukemia in mice. The antitumor action of fucoidan from Eisenia bicyclis appears to be exerted through enhanced NK cell activity. it was also reported that mekabu fucoidan augments type1 Th1 cell responses as well as the production of INF-γ and IL 12. Moreover, fucoidan inhibited the growth of subcutaneously inoculated P-388 leukemia cells in mice.

8. The present study examines the antiallergic activities of mekabu fucoidan. It was examined how mekabu fucoidan influences the Th1/Th2 balance in a Th2-biased immune system. We induced Th2 responeses in mice by challenge with ovalbumin and then examined whether mekabu fucoidan suppresses IgE levels and downregulates Th2 cytokines in this model. The results revealed that mekabu fucoidan suppresed the Th2 immune response. In Present Study

10. Materials and Methods Mekabu Fucoidan Evaporated and lyophilized Filter Supernatant was passed Neutralized with 3N NaOH Ultrafiltered Centrifuge 12227g, 10min at 4 ℃ Extract 0.15N HCl at 22 ℃ for 15h Wash twice 80% ethanol at 80 ℃ for 3h Obtained from sporophyll of U.pinnatifida and Milled

11. Materials and Methods Animals Male BALB/c mice at 6 weeks of age specific pathogens free-conditions

12. Materials and Methods Experimental Design Fucoidan Chicken OVA and alum Aerosol of OVA BALB/c mice Saline Chicken OVA and alum Aerosol of OVA Control mice

13. Fucoidan 50mg/kg in 0.1ml For 4 days 0.5ml PBS ( chicken OVA 50μg And alum 1mg) In mouse chamber ( aerosol of OVA 100mg/ml for 30min) ① Blood sample ② Bronchoalveolar lavage fluid(BALF) ③ Fixed lungs After 1 week After 8 and 9 days After 24h

14. Materials and Methods ELISA for IgE and Anti-OVA Antibody 96 well microtiter plate Anti-IgE Ab Anti-OVA Ab Serum sample Anti-IgE Ab

15. Materials and Methods Detection of Eosinphills in BALF Using a cytospin - BALF(100μl) smear on glass slide - 3000rpm for 5min - fixed in 95% ethanol Diff-Quick stain Count -under an optical microscope -number per 1000 effusion cells

16. Materials and Methods Histopathological Examination Lung tissue - fixed 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4) - embedded in paraffin - H&E stain or PAS stain Mucosal material and leukocyte infiltlation - histologically examined Goblet cells in airway epithelium -PAS stain -count Statistical Methods The result are expressed as means ± standard error of the mean.

17. Result Induction of IL-4, IL-5, IL-13 and IFN-γ in BALF

18. Result Total IgE and OVA-Specific IgE in blood serum

19. Result Presence of eosinophils in BALF(Diff-Quick staining) Control mice Mice treated with mekabu fucoidan

20. Result Number of Eosinophils in BALF Eosinophils decreased

21. Result Histopathological Change of OVA-Specific Allergic Airways Disease

22. Result Number of Goblet cells among bronchial epithelial cells Goblet cells decreased

23. Th1 Fucoidan IFN-γ IL-12 IFN-γ IL-12 Anti-Tumor Action Anti-Tumor Action Th2 IL-4 IL-13 IL-4 IL-13 Hyper Air inflammation Hyper Air inflammation Discussion Future studies should compare the immunosuppressive actions of mekabu fucoidan with those of an antiallergic agent, as well as in combination with other agents.

24. Discussion We plan to evaluate the effects of oral mekabu fucoidan in mice.

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