| Summary: | 碩士 === 國立清華大學 === 生命科學系 === 89 === Toxin-r is the major compound in the venom of African spitting cobra,Naja nigricollis; it includes in the cardiotoxin family, and could include various effect such as the depolarization of neuron and muscle,hemolysis, and also involve in the interruption of the equilibrium of ionchannel, and exhibit various cytotoxicity effects. Defensin, a protein which have the ability in killing bacterial, and in regulating the function of immunocyte. We found that cardiotoxin have a high similarity in structure with defensin,and since their structure resemble to each other,we wonder whether they also exhibit some similarity in their function.First at all, in the virulence experiment done in our lab have confirm the result of the cardiotoxin have no ability in killing bacteria. But in the regulation of immunocyte functions experiment,we found out although toxin-r at high concentration show a cytotoxicity effect toward neutrophils, but low concentarion of toxin-r in reversal could actiate chemotaxis,chemokinesis, NADPH oxidase, and actin polymerization in neutrophils; which the degree of activation is proportional to the concentration of toxin-r, this observation may be due to the actin polymerization induced by the toxin-r. The signal transduction pathway of various activations induce by toxin-r in neutrophils is not yet observed, but here we can rule out the possibility of the involvement of calcium ion in signal transduction, we also perform that actin polymerization induced would decrease in the presence of the signal transduction inhibitor inside the cell.
1.1.心臟毒素簡介 5
1.1.1. 蛇毒 5
1.1.2. 心臟毒素 6
1.1.3. Defensin 7
1.2. 嗜中性白血球簡介 8
1.2.1. 嗜中性白血球 8
1.2.2. 化學趨化性 9
1.2.3. NADHP氧化 9
1.2.4. 肌動蛋白 11
1.2.5. 鈣離子 14
1.2.6. cAMP 16
2. 實驗材料及方法 23
2.1. 材料 23
2.2. 方法 24
2.2.1. 蛇毒心臟毒素的製備 24
2.2.2. 嗜中性白血球分離 25
2.2.3. 細胞毒性測定 27
2.2.4. 化學趨化功能的測定 28
2.2.5. 過氧化物濃度測定 30
2.2.6. 絲狀肌動蛋白含量測定 31
2.2.7. 細胞內鈣離子濃度測定 34
2.2.8. cAMP對肌動蛋白聚合反應與化學趨化性的影響 35
3.結果 36
3.1. Tγ對嗜中性白血球的細胞毒性 36
3.2. Tγ對化學趨化性的影響 36
3.3. Tγ對NADPH氧化的影響 37
3.4. Tγ活化球狀肌動蛋白的聚合反應 38
3.5. 鈣離子未參與訊息傳遞 39
3.6. cAMP對嗜中性白血球功能的影響 40
4. 討論 53
5. 參考文獻 58
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