Kang Kyung-Hwa, Lee Kyung-Hee, Yoon Hyun-Min, Jang Kyung-Jeon, Song Chun-, Kim Cheol-Hong
Department of Oriental Physiology, Dong-Eui University College of Oriental Medicine and Research Institute of Oriental Medicine, Busan, Korea.
Department of Acupuncture & Moxibustion, Dong-Eui University College of Oriental Medicine and Research Institute of Oriental Medicine, Busan, Korea.
J Pharmacopuncture. 2012 Dec;15(4):32-41. doi: 10.3831/KPI.2012.15.015.
Rehmannia glutinosa pharmacopuncture solution (RGPS) was investigated to determine both its anti-allergic inflammatory effects on mast cells and its detailed mechanism of actions.
We investigated whether RGPS suppress cytokines, enzymes, FcεRI expression and FcεRImediated signaling in RBL-2H3 cells stimulated with anti-DNP IgE/DNP-HSA. The suppressive effects of RGPS on the levels of cytokines such as IL-1β, IL-6 and GM-CSF were measured using emzyme-linked immunospecific assay (ELISA). The mRNA expression levels of cytokines, enzymes (HDC2, COX-1, COX-2 and 5LO) and FcεRI αβγsubunits were measured using reverse transcription polymerase chain reaction (RTPCR) method. The activation of FcεRI-mediated signaling was examined using Western blot analyses.
RGPS suppressed production of proinflamm-atory cytokines (IL-1β, IL-6, and GM-CSF) in stimulated RBL-2H3 cells significantly (p< 0.05). RGPS also suppressed mRNA expression of inflammatory enzymes (HDC2, COX-1, COX-2, 5LO). In addition, mRNA expression levels of FcεRIα, FcεRIβand FcεRIγ were lowered by treatment with RGPS. Finally, RGPS prevented phosphrylation of Lyn, Syk, LAT, Gab2, PLC γ1/2, PI3K, Akt, cPLA2 and IκBα.
RGPS effectively suppresses mast cell activations such as degranulation and inflammatory response via down-regulation of the FcεRI-mediated signaling pathways in IgE/Ag-stimulated mast cells.
研究熟地黄药刺溶液（RGPS）对肥大细胞的抗过敏炎症作用及其详细作用机制。
我们研究了RGPS是否能抑制抗DNP IgE/DNP-HSA刺激的RBL-2H3细胞中的细胞因子、酶、FcεRI表达和FcεRI介导的信号传导。使用酶联免疫特异性测定（ELISA）测量RGPS对IL-1β、IL-6和GM-CSF等细胞因子水平的抑制作用。使用逆转录聚合酶链反应（RTPCR）方法测量细胞因子、酶（HDC2、COX-1、COX-2和5LO）和FcεRIαβγ亚基的mRNA表达水平。使用蛋白质印迹分析检查FcεRI介导的信号传导的激活情况。
RGPS显著抑制了抗DNP IgE/DNP-HSA刺激的RBL-2H3细胞中促炎细胞因子（IL-1β、IL-6和GM-CSF）的产生（p<0.05）。RGPS还抑制了炎症酶（HDC2、COX-1、COX-2、5LO）的mRNA表达。此外，用RGPS处理可降低FcεRIα、FcεRIβ和FcεRIγ的mRNA表达水平。最后，RGPS阻止了Lyn、Syk、LAT、Gab2、PLCγ1/2、PI3K、Akt、cPLA2和IκBα的磷酸化。
RGPS通过下调IgE/Ag刺激的肥大细胞中FcεRI介导的信号通路，有效抑制肥大细胞的活化，如脱颗粒和炎症反应。
