The Experimental Study Of Liu-shen-wan On Inducing HL-60 Cell Apoptosis With Chinese Herbs Serum Pharmacology Method

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.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.