【作者】 罗英儒；

【导师】 叶春玲；

【作者基本信息】 暨南大学 ， 药理学， 2002， 硕士

【摘要】 本研究采用血清药理学方法，以人白血病细胞HL60、T、B淋巴瘤细胞及正常人胚肺二倍体细胞为靶细胞，通过台盼兰拒染试验、流式细胞仪分析、DNA片断凝胶电泳、免疫组织化学(SP法)等观察中华眼镜蛇毒兔血清体外对白血病和淋巴瘤细胞增殖及诱导凋亡的影响，并探讨其可能的作用机制。结果显示：(1)与空白对照组比较有显著性差异，中华眼镜蛇毒兔血清在体外显著抑制HL60细胞的生长，其抑制作用随着剂量的加大而增强，高剂量(7.5mg／ml)的抑制效果相近于阿霉素(5μg／ml)。而对Jurkat和Raji细胞，当剂量达到15mg／ml时，中华眼镜蛇毒兔血清才表现出同样强度的抑制作用。(2)中华眼镜蛇毒兔血清对正常人胚肺二倍体细胞增殖无抑制。(3)经DNA电泳、流式细胞仪分析显示：中华眼镜蛇毒兔血清7.5mg／ml与HL60细胞共同培养48小时可诱导其细胞凋亡增加。Jurkat及Raji细胞则在15mg／ml剂量时出现细胞凋亡率的增加。(4)中华眼镜蛇毒兔血清作用48小时后，HL60细胞bcl-2、C-myc基因蛋白表达下调。上述结果表明：(1)中华眼镜蛇毒兔血清体外能显著抑制白血病和淋巴瘤细胞的生长，此作用可能与诱导细胞发生凋亡有密切关系。而对正常细胞，中华眼镜蛇毒兔血清未表现出明显的细胞毒作用。(2)人白血病细胞HL60对中华眼镜蛇毒兔血清的敏感性较Jurkat、Raji淋巴瘤细胞高。(3)中华眼镜蛇毒兔血清可使HL60细胞bcl-2、C-myc基因蛋白表达下调，可能与其抑制细胞增殖及诱导细胞凋亡密切相关。更多还原

【Abstract】 In order to explore the effects and mechanism of Chinese cobra venom serum of rabbit (CVSR) on proliferation and apoptosis in leukemia and lymphoma cells in vitro, serologic pharmacological test was used. The effects were observed by means of trypan blue dye exclusion test, flow cytometry analysis, agrose gel electrophoresis and immunohistochernistry. The results showed that: (1) HL60 cells were inhibited significantly by the dosage of 2.5mg/ml, 5.0 mg/ml, 7.5mg/ml of CVSR. At the same dosage, there is no influence of CVSR on proliferation of human embryo lung diploid cells. Furthermore, the Jurkat and Raji cells showed the same inhibition when the dosage was added to 15 mg/ml. (2) By flow cytometry analysis, we found that the apoptosis rate of HL60 cells was significantly increased by the CVSR. A characteristic DNA "ladder" was detected by using agarose gel electrophoresis. (3) Bcl-2 and C-myc gene protein expression of HL60 cells was down-regulated. The results suggested that: (1) CVSR inhibited the growth of leukemia and lymphoma cells but had no cytotoxin on human normal cells in vitro. (2) HL60 cells was more sensitive than Jurkat and Raji cells to the CVSR. (3) The gene protein expression of bcl-2 and C-myc on HL60 cells down-regulated by CVSR was probably related to apoptosis.更多还原