紫蘇萃取物抑制歐洲室塵蹣過敏原Der p 2誘發的支氣管上皮細胞BEAS 2B發炎反應之機制研究

歐洲室塵蹣第二類群過敏原(Dermatophagoides pteronyssinus grop 2(Der p 2)為不具有蛋白水解酶特性的主要塵蹣過敏原之一，這類過敏原對呼吸道上皮細胞的潛在影響相關的知識有限，不過有研究顯示，利用重組蛋白Der p 2去刺激小鼠上呼吸道，會導致小鼠產生特異性的免疫球蛋白IgE以及呼吸道發炎現象，可藉由活化細胞核轉錄因子 NF-κB，以及活化促分裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)，導致支氣管上皮細胞釋放細胞激素(cytokines)與趨化激素(chemokines) 。在本實驗中，由於我們由之前的文獻已知，DP2會誘發小鼠呼吸道發炎，且可以活化支氣管上皮細胞MAPK以及NF-κB活化，使小鼠發炎相關的cytokines與chemokines表現量增加；而紫蘇葉提取物也已被研究具有抗發炎，抗過敏，抗氧化活性具有治療潛力。於是我們想進一步研究紫蘇多酚對於塵蹣過敏原所引起的支氣管過敏性炎症之影響。我們首先由中藥行購入乾燥紫蘇葉進行紫蘇多酚PFE的萃取，接著以PFE預處理正常呼吸道上皮細胞BEAS-2B，之後再以DP2刺激，觀察細胞是否表現與過敏或發炎反應相關的cytokines與chemokines，以及此現象是否能被PFE所抑制。由結果可以得知，當以塵蹣過敏原DP2刺激正常呼吸道上皮細胞，會使MAP Kinase磷酸化增加，增加NF-κB轉位至細胞核內，使細胞內產生與發炎相關的cytokines與chemokine，而先以紫蘇多酚預處理過，在以DP2刺激BEAS 2B細胞，的確可以抑制細胞MAPK活化以及NF-κB的核轉位(nuclear translocation)。實驗結果並進一步證實PFE是透過抑制IκBα降解，使NF-κB無法進行核轉位，進而使細胞發炎相關的cytokines與chemokines不會被表現。上述研究結果顯示紫蘇多酚PFE可以有效抑制由過敏原所引起的呼吸道上皮細胞發炎反應。

關鍵字

歐洲屋塵蹣過敏原第二群

並列摘要

The Perilla frutescens Britt. (PF), a traditional Chinese medicinal herb, has been used in China for centuries to treat various diseases including bacterial and fungal infections and allergic diseases.House dust mites are well-known as a source of indoor aeroallergens and for causing allergic airway diseases. In this study, we investigated the anti-allergic effect of methanol extract of PF leaf (PFE) on Dermatophagoides pteronyssinus 2 (DP2)-stimulated bronchial epithelial cell line (BEAS-2B). Pro-inflammatory cytokines mRNA expression was analyzed by RT-PCR and quantitative real-time PCR. Activation of kinase cascades were investigated by immunoblot. Our results revealed that PFE significantly reduced the DP2-induced mRNA expression of IL-4, IL-5, IL-6, IL-8, IL-13, MCP-1 and GM-CSF in a dose-dependent manner. A mechanism based kinase cascades analysis showed that PFE suppressed the DP2- induced phosphorylation of mitogen-activated protein kinase (MAPK) including extracellular signal-regulated kinase1/2 (ERK1/2) , P38 MAPK(p38) and c-Jun N-terminal kinase (JNK).Additionally,we also found that PFE inhibited the translocation of NF-κB/p65 into nucleus which may result from the suppression of IκBα degradation. Taken together, these findings indicate that PFE significantly alleviates DP2-induced inflammatory responses, which may attribute to suppression of MAPK signaling and nuclear of NF-κB, suggesting that PFE should be beneficial to treatment of allergen-induce inflammation.

並列關鍵字

house dust mite allergen Der p 2

參考文獻

[1] Platts-Mills T, Vervloet D, Thomas W, Aalberse R, Chapman M.Indoor allergens and asthma: report of the Third International Workshop. J Allergy Clin Immunol 1997; 100:S2–24.

[2] Custovic A, Taggart S, Francis H, Chapman M, Woodcock A.

Exposure to house dust mite allergens and the clinical activity of asthma. J Allergy Clin Immunol 1996; 98:64–72.

[3] Ueda H, Yamazaki C, Yamazaki M. Luteolin as an anti-inflammatory and anti-allergic constituent of Perilla frutescens.Biol Pharm Bull 2002; 25:1197–202.

[6] Menzies-Gow A, Robinson DS. Eosinophils,eosinophilic cytokines(interleukin-5), and antieosinophilic therapy in asthma. Curr Opin Pulm Med 2002; 8:33–8.

國際替代計量

紫蘇萃取物抑制歐洲室塵蹣過敏原Der p 2誘發的支氣管上皮細胞BEAS 2B發炎反應之機制研究

全文下載

主題瀏覽