探討葡萄籽萃取物具有抗過敏作用是藉由調控第一型血基質氧化酶的表現

Investigation of the potential anti-allergic properties of grape seed extract via up-regulation of heme oxygenase-1 in RBL-2H3 mast cells

洪美惠(Mei-Huei Hung)

指導教授 ： 陳炳宏

高雄醫學大學/生命科學院/生物科技學系碩士班/碩士(2011年)

https://doi.org/10.6832/KMU.2011.00106

摘要

過敏為全世界性的疾病，隨著社會經濟的發展，過敏性疾病的發生率越來越高，像是氣喘、過敏性鼻炎與異位性皮膚炎。而過敏的發生是環境中的抗原再次與體內免疫球蛋白 E結合後，藉由細胞表面上的FcεRI與其結合所引起的免疫反應，造成肥大細胞與嗜鹼性球細胞的活化並產生去顆粒作用，釋放出許多發炎媒介物質，像是組織胺、趨化因子、血小板活化因子、白三烯素以及前列腺素。本研究目的為如果可以降低或抑制肥大細胞表面上的FcεRI含量，或者是影響抑制過敏發生的生理反應途徑，即可能發展抗過敏之藥物。而目前研究發現，生物類黃酮激素具有抗氧化作用，可從許多植物萃取而得。其中，葡萄籽萃取物含有大量的類黃酮激素-寡聚合物原花青素。為了探討葡萄籽萃取物是否能減緩或抑制過敏反應的發生，利用流式細胞儀與即時定量聚合酶連鎖反應分析經葡萄籽萃取物處理後的RBL-2H3肥大細胞。實驗結果顯示，細胞表面上FcεRI的含量與細胞內FcεRI mRNA的表現量明顯減少。進一步的利用RT-PCR、即時定量聚合酶連鎖反應與西方墨點法分析葡萄籽萃取物可能影響過敏反應發生的途徑。實驗結果顯示，經葡萄籽萃取物處理後，第一型血基質氧化酶的表現會被調升。而調控第一型血基質氧化酶的轉錄因子-Nrf2可由西方墨點法實驗結果看出，其Nrf2可由細胞質轉位進入細胞核，以調控第一型血基質氧化酶的表現。綜合以上實驗結果，本論文證明葡萄籽萃取物可能是透過調升第一型血基質氧化酶的表現而達到降低或者是抑制過敏反應的發生。

關鍵字

葡萄籽萃取物 ； 肥大細胞 ； 過敏 ； 第一型血基質氧化酶

並列摘要

Allergy is a global disease, and due to socio-economic development, there is a higher incidence of the allergic diseases such as asthma, allergic rhinitis and atopic dermatitis. Allergy reaction occurs as a result of the antigen in the environment binds to immunoglobulin E followed by its binding with FcεRI upon the cells. This results in the activation and degranulation of mast cells and basophils and the release of inflammatory mediators including histamine, chemotactic factor, platelet activating factor, leukotriene, and prostaglandin. The aim of this study was to examine whether grape seed extracts (GSE) could inhibit the expression of FcεRI on mast surface, or influence the allergic pathway, which may develop anti-allergic drugs. Current studies have found that bioflavonoids possess anti-oxidant effect, and can be extracted from many plants. GSE contain a large amount of bioflavonoids, and among those, oligomeric proanthocyanidins (OPCs) is the main one. GSE have high antioxidant activities and free radical scavenging properties that can be beneficial to the human health. In recent studies, it was suggested that the consumption of fruits and vegetables can help reducing the risks of various cancers including breast cancer, lung cancer, and colorectal cancer. To investigate whether GSE can inhibit the allergic reaction, flow cytometry and real-time polymerase chain reaction were used to analyze GSE treated RBL-2H3 mast cells. The results demonstrated that FcεRI on the cell surface increased and intracellular mRNA levels expression was significantly enhanced. For further investigation, RT-PCR, real-time PCR and western blot analysis were used to test the allergic pathway affected by GSE and increased expression of heme oxygenase-1 (HO-1) in the cells treated with GSE was observed. In addition, western blot analysis demonstrated the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2), the transcription factor of heme oxygenase-1, into the cell nuclease to regulate HO-1 expression.
In conclusion, the thesis proves that GSE inhibit allergic reactions through up-regulating HO-1 expression and by the cytoproduction of CO and bilirubin.

並列關鍵字

grape seed extract ； mast cells ； allergy ； heme oxygenase-1

參考文獻

1. 黃群峰. 2006. 台灣東部地區學童過敏疾病盛行率調查及過敏原研究-以花蓮縣玉里鎮全鎮國小學童為研究對象. In 國立中山大學 生物醫學研究所 碩士論文.

2. Gould, H. J., and B. J. Sutton. 2008. IgE in allergy and asthma today. Nat Rev Immunol 8:205-217.

3. Poggianella, M., M. Bestagno, and O. R. Burrone. 2006. The extracellular membrane-proximal domain of human membrane IgE controls apoptotic signaling of the B cell receptor in the mature B cell line A20. J Immunol 177:3597-3605.

4. Bestagno, M., L. Vangelista, P. A. Mandiola, S. Mukherjee, J. Sepulveda, and O. R. Burrone. 2001. Membrane immunoglobulins are stabilized by interchain disulfide bonds occurring within the extracellular membrane-proximal domain. Biochemistry 40:10686-10692.

5. Takai, T. 2002. Roles of Fc receptors in autoimmunity. Nat Rev Immunol 2:580-592.