摘要
屬於核醣核酸酵素A基因家族的人類嗜酸性陽離子蛋白質(ECP)，經由受到活化的嗜伊紅性白血球釋出，具有多重的生物功能其中含有對於神經細胞的毒性，抗原核生物以及抗寄生蟲生長的毒性，對於不同的細胞型態有不同的細胞毒性特異性，而至今我們對於它的毒性機制尚未有完全的瞭解；在因過敏所引起的發炎反應，如氣喘症候中，人類嗜伊紅性白血球受到吸引移動到發炎反應區域，而受到活化後釋出內含顆粒性蛋白質引起的發炎組織中細胞凋亡現象。因此我們選用了人類呼吸道上皮細胞株作為試驗的對象，並且以鎳親和力管住純化了重組嗜酸性陽離子蛋白質作為實驗的材料，實驗當中，我們發現在細胞型態上，流式細胞儀的檢測，以及DNA片段的分析中，細胞發生了細胞凋亡；根據前人的研究，人類嗜酸性陽離子蛋白質的核醣核酸酵素活性對於神經細胞毒性為其所必須，相反的對於抗原核生物，抗寄生蟲生長的活性卻非必要，因此我們利用了點突變技術以降低了人類嗜酸性陽離子蛋白質的核醣核酸酵素活性，以利後續對於人類嗜酸性陽離子蛋白質的毒性機制研究。

Abstract
Eosinophil cationic protein (ECP) is one of the proteins in the ribonuclease A (RNase A) superfamily that have developed various biological properties related to the function of eosinophils. ECP secreted by activated human eosinophils that has anti-parastic, antibacterial, and neurotoxic activities. It is a potent cytotoxic molecule, and the mechanism remains unknown. Reported study suggest that growth inhibition by ECP is dependent on cell type and is cytostatic. In allergic inflammatory process is associated almost invariably with tissue damage and healing. The respiratory epithelium is an essential target of the inflammatory attack by eosinophils in asthma. Bronchial epithelial cells show morphological characteristics of apoptosis mediated by activated eosinophils. To determine the cytotoxic effect of ECP on BEAS-2B cells line. A method for producing recombinant ECP in a prokaryotic expression system was devised. Periplasmic isolates from induced bacterial transfectants contained enzymatically active ECP; micromolar concentration of ECP were shown to be toxic for BEAS-2B cells line. As the data shown, we find that the ECP treatment on BEAS-2B cells has apoptotic effect. The result match to apoptosis induced in inflammatory tissue and allergic disease by activated eosinophils.

Due to report study, the RNase activity of ECP companies to its neurotoxic activities, but opposite to its anti-parasite and antibacteridal activity. During our experiment, we decreased the RNase activity of ECP by site direct mutation for further test.

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