RBL-2H3 비만세포와 OVA/alum 감작 생쥐에서 淸心補血湯의 항알레르기 효과
Anti-allergic Effects of Chungshimbohyeoltang in RBL-2H3 Mast Cells and OVA/alum Sensitized Mice
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Objectives: 청심보혈탕의 항알레르기의 효과와 기전을 규명하고자 하였다. Methods: 청심보혈탕의 항알레르기 효과를 알아보기 위하여 RBL-2H3 비만세포를 이용하여 Real-Time PCR과 ELISA로 IL-4와 IL-13를 분석하였고, western blot으로 GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos, c-Jun, NF-κB p65 등의 전사인자의 발현을 관찰하였다. 그리고 OVA/alum을 감작한 생쥐 모델에서 OVA-specific IgE 생성량과 Th2 사이토카인인 IL-4와 IL-13 생성량을 측정하였다. Results: RBL-2H3 비만세포에서의 항알레르기 효과를 살펴본 결과 청심보혈탕은 생쥐 비만세포주의 모든 농도에서 세포독성이 나타나지 않았다. Real-Time PCR를 분석한 결과 IL-4와 IL-13 mRNA 유전자 발현을 억제하였다. ELISA 측정 결과 청심보혈탕은 IL-4와 IL-13 단백질 생성을 억제하였다. Western blot 분석 결과 청심보혈탕은 GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos, NF-κB p65 단백질 발현을 억제하였다. OVA/alum 감작 생쥐에서의 항알레르기 효과를 살펴본 결과 청심보혈탕은 혈청 중 OVA-specific IgE 생성을 억제하였고 비장세포 배양 상층액에서의 IL-4와 IL-13 생성을 억제하였다. Conclusions: 淸心補血湯은 알레르기 염증 반응시 Th2 세포가 분비하는 IL-4와 IL-13의 생성을 억제하고, 비만세포의 특이적 전사인자인 GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos 및 NF-κB p65를 억제함으로써 IL-4와 IL-13 생성을 억제한다.
Abstract
Objectives The suppressive effect of CSBHT has been mysterious. Thus, the present study is designed to investigate the suppressive effect and its mechanism. Methods To investigate the anti-allergy effect from ChungShimBoHyeolTang(CSBHT), RBL-2H3 cell was used and examined by Real-Time PCR, and IL-4 and IL-13 from RBL-2H3 was examined by ELIS. In addition, GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos, c-Jun, NF-κB p65 transcription factors of RBL-2H3 mast cell were examined by Western Blotting. Also, OVA/alum-sensitized mice were orally administrated CSBHT and serum OVA-specific IgE production, IL-4, and IL-13 production in splenocytes supernatant were examined. Results As a result of treating with CSBHT extract, RBL-2H3 mast cells significantly suppressed the IL-4 and IL-13 mRNA expression and IL-4 and IL-13 production. Western blot analysis of transcription factors involving IL-4 and IL-13 expression also revealed a prominent decreases of mast cell's specific transcription factors including GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos, and NF-κB p65. Also, examining the mice, administration of CSBHT suppressed the amount of OVA-specific IgE in OVA/alum-sensitized mice and IL-4 and IL-13 production in splenocytes supernatant. Conclusions The study suggested that the anti-allergic activities of CSBHT suppresses IL-4 and IL-13 production from the Th2 cytokines by suppressing transcription factors as GATA-1, GATA-2, NFAT-1, NFAT-2, c-Fos and NF-κB p65 in mast cells.
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