沙利度胺对于M7细胞作用的实验研究.docVIP

精品论文 参考文献 沙利度胺对于M7细胞作用的实验研究 冯 林 　　泸州医学院附属医院血液科 四川泸州 646000 　　【摘 要】目的：研究沙利度胺对于M7细胞（巨核细胞白血病细胞）的作用。方法：用Sigma公司生产的沙利度胺原料药。以0.1%DMSO（二甲基亚砜）为阴性对照组，以M7细胞为空白组，以不同浓度的沙利度胺作用于M7细胞，用CCK-8方法（细胞计数试剂盒方法）观察不同浓度不同时间沙利度胺对于M7细胞的增值抑制作用；以不同浓度沙利度胺分别作用M7细胞24h, 48h, 72h后，用ELISA方法检测上清液中VEGF（血管内皮生长因子）的浓度；以不同浓度沙利度胺分别作用M7细胞24h, 48h, 72h后，经HE及DAPI（二脒基苯基吲哚）染色后，用光学显微镜及荧光显微镜观察M7细胞形态变化，并应用统计学分析实验结果。结果：沙利度胺可明显抑制M7细胞分泌VEGF，并且抑制作用随着沙利度胺浓度及作用时间而增强；在CCK-8实验中，各组M7细胞与空白组相比，发现沙利度胺对M7细胞无明显抑制作用；光学显微镜观察不同时间段不同组，各组M7细胞与空白组相比较，细胞生长较好，呈圆形，细胞核大，核膜清晰。荧光显微镜下，未见明显凋亡小体。结论：1.沙利度胺能够明显抑制M7细胞分泌VEGF，并且呈时间浓度依赖性。2.沙利度胺对于M7细胞没有明显抗增殖效应及诱导其凋亡的作用。 　　【关键词】沙利度胺；VEGF；M7细胞; 凋亡小体；DAPI染色 　　【中图分类号】R52.5 【文献标识码】B 【文章编号】1674-8999（2015）9-0566-02 　　【Abstract】Objective：To research on the impact of thalidomide on M7 cell line. Methods：Thalidomide used as crude drug was bought from Sigma company. Take 0.1%DMSO as the negetive control group, and M7 cells as blank group, to treat M7 cell with thalidomide of different concentration and check the proliferative inhibition rate of M7 cell by CCK-8; to treat M7 cell with thalidomide of different concentration for 24hr ,48hr and 72hr respectively and check the VEGF concentration of supernatant by Elisa; to treat M7 cell with thalidomide of different concentration for 24hr ,48hr and 72hr respectively and check the forms and apoptotic state of M7 cells by light microscope and fluorescent microscope and analyze the experiment results by statistics. Results：Thalidomide can remarkablely inhibit the VEGF secreted by M7 cell and the inhibition increases as the time and concentration go by. In CCK-8 test, to compare each subject group with blank group, we find thalidomide has no remarkable inhibitory action on M7 cell. To compare each subject group of different times with blank group by light microscope and fluorescent microscope, we find cells grow well with round shape, large nucleus and limpid nuclear membrane with HE staining and there is no apparente apoptotic body with DAPI staining. Dis