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机构地区:[1]北京医科大学基础医学院药理学系
出 处:《中国药理学通报》1999年第1期27-30,共4页Chinese Pharmacological Bulletin
摘 要:目的探讨灵芝菌丝体粗总多糖(MGLP1)、总多糖(MGLP2)作用的小鼠腹腔巨噬细胞培养上清对HL-60细胞凋亡的影响。方法MGLP1、MGLP2作用于小鼠腹腔巨噬细胞,生物法检测培养上清中TNF-α的活性;再将共培养上清(MGLP1-ΜΦ-CM、MGLP2-ΜΦ-CM)作用于HL-60细胞,MTT法检测MGLP1-ΜΦ-CM、MGLP2-ΜΦ-CM对HL-60细胞增殖的抑制作用;琼脂糖凝胶电泳法和流式细胞检测法定性定量检测它们对HL-60细胞凋亡的诱导作用。结果MGLP1、MGLP2作用于腹腔巨噬细胞后,可以明显增强培养上清中TNF-α的活性;其共培养上清可以明显地抑制HL-60细胞的增殖并诱导该细胞凋亡(P<0.01)。结论MGLP1、MGLP2可以通过小鼠腹腔巨噬细胞,促进TNF-α分泌,诱导HL-60细胞凋亡。AIM The present study was to investigate the effects of the conditioned medium of polysaccharides isolated from mycelia of Ganoderma lucidum (MGLP1 and MGLP2) activated macrophages (MGLP1 MΦ CM, MGLP2 MΦ CM) on HL 60 cell apoptosis. METHODS TNF α activity was determined by biological method. The in vitro effects of MGLP1 MΦ CM, MGLP2 MΦ CM on the proliferation of HL 60 cells were evaluated by MTT assay. DNA gel electrophoresis and flow cytometric analysis were use to determine the apoptotic cells. RESULTS The influence of MGLP1 and MGLP2 on TNF α activity in the peritoneal macrophage cell conditioned media (MGLP1 MΦ CM, MGLP2 MΦ CM) displayed enhancement in concentration dependent manner (12 5 ̄200 mg·L -1 ). When MGLP1 and MGLP2 (50 ̄200 mg·L -1 ) treated peritoneal macrophages for 2 days, the MGLP1 MΦ CM and MGLP2 MΦ CM were found to suppress the proliferation of HL 60 cell line; DNA gel electrophoresis showed that treatment with MGLP1 MΦ CM and MGLP2 MΦ CM (20%) induced HL 60 cell apoptosis. Flow cytometric analysis revealed that 23 75% (MGLP1 MΦ CM) and 24 68% (MGLP2 MΦ CM)apoptotic cells were seen while in control culture (N MΦ CM) there were only few (5 24%) apoptotic cells CONCLSION MGLP1 and MGLP2 induced HL 60 cell apoptosis by enhancing macrophages activity and promoting TNF α production.
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