灵芝菌丝体多糖通过增强小鼠巨噬细胞功能诱导HL-60细胞凋亡

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。

Polysaccharides isolated from mycelia of Ganoderma lucidum induced on HL 60 cell apoptosis by enhancing macrophage activity

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.