【作者】 黄振东；

【导师】 戴锡孟；

【作者基本信息】 天津中医学院 ， 中医内科学， 2003， 硕士

【摘要】 目的：1 研究六神丸含药血清的制备，探讨六神丸血清药理学的实验方法。2 观察六神丸含药血清诱导白血病细胞凋亡，探讨六神丸治疗急性白血病的部分机制。方法：1 分不同的给药剂量、给药天数和不同的采血时间制备含药血清。2 用原子吸收法测定血清砷含量；薄层色谱法定性检测六神丸含药血清中蟾酥成分。3 用MTT法、胎盘蓝染色法观察六神丸对HL-60细胞的增殖抑制作用，确定最佳血清制备方案。4 用吉姆萨染色法观察细胞凋亡的形态学变化，流式细胞仪、原位末端标记法（TUNEL）定量检测细胞凋亡程度。结果：1 血清砷含量随着给药剂量、给药天数增加和采血时间的延长而升高；薄层色谱表明六神丸含药血清中有蟾酥成分。2 六神丸含药血清对白血病细胞的抑制率，随给药剂量、天数增加和采血时间的延长而升高3 吉姆萨染色、流式细胞仪检测、原位末端标记检测均证明六神丸含药血清能诱导HL-60白血病细胞凋亡。结论：1 中药血清药理学含药血清的制备应区分不同的中药，采用相应的方案。2 六神丸含药血清能诱导细胞凋亡，这可能是六神丸治疗白血病的机制之一。更多还原

【Abstract】 Purpose1. To study the making method of medicine serum of Liu-Shen-Wan, to explore Liu-Shen-Wan serum pharmacology method.2. To observe medicine serum of Liu-Shen-Wan inducing leukaemia cell apoptosis and to explore the partial mechanism of liu-shen-wan in treating acute leukaemiaMethod1.To produce medicine serum of Liu-Shen-Wan with different dose, different time of pouring medicine, different time of blood sampling.2. To measure content of arsenic in serum with atomic absorption method, to qualitative analysis toadfish in medicine serum of Liu-Shen-Wan with thin layer chromatography.3. MTT and trypan blue assay detects the growth inhibition effect of Liu-Shen-Wan on HL-60 cells and to make certain the best scheme of making method of medicine serum of Liu-Shen-Wan.4. Gimsa dying assay observes morphology changes of apoptosis .In situ end-labeling method (POD) and flow cell machine quantificationally probes apoptosis phenomena.Results1. Content of arsenic in serum raises with dose adding, time of pouring medicine and time of blood sampling prolonging. Thin layer chromatography indicates that there is toadfish in medicine serum of Liu-Shen-Wan.2. The suppressive rate of medicine serum of Liu-Shen-Wan on HL-60 cell raises with adding of dose and time of pouring medicine and prolonging of blood sampling.3. Gimsa dying assay, in situ end-labeling method (POD) and flow cell machine all exhibits: HI-60 cells dealed with Liu-Shen-Wan medicine-serum present typical apoptosis morphology.<WP=7>Conclusions1. To use the corresponding scheme of producing medicine serum for different chinese traditional medicine.2. Liu-Shen-Wan medicine-serum can induce apoptosis of HL-60 cell. It is one of mechanisms in treating leucemia by Liu-Shen-Wan.更多还原