香菇多糖对小鼠巨噬细胞活化作用.docVIP

香菇多糖对小鼠巨噬细胞活化作用 　【摘要】 目的 探讨香菇多糖（LNT）对机体的免疫系统刺激活性。方法 （1）615纯系小鼠静脉注射LNT，测定巨噬细胞吞噬功能；（2）肿瘤坏死因子（TNF）的测定；取传代后3～4天L615小鼠白血病细胞检测细胞毒；（3）荧光染色和DN A电泳观察细胞凋亡。结果 LNT能够诱导体外培养的巨噬细胞、脾细胞和胸腺细胞的培养上清液中TNF含量显著增加。诱导后的巨噬细胞上清对L615白血病细胞细胞毒活性由对照的11．4%增高到66．2％。且呈LNT剂量依赖性，作用后的白血病小鼠细胞体积显著缩小，核发生固缩，呈现凋亡细胞特征。结论 LNT活化后的巨噬细胞对肿瘤细胞有显著的细胞毒作用，其机制与诱导巨噬细胞产生TNF而介导肿瘤细胞凋亡有着密切的关系 【关键词】 香菇多糖 巨噬细胞 肿瘤坏死因子 细胞凋亡 【Abstract】 Objective To research one of its mechanisms mediated by the activation of macrophages in murine cavity induced by lentinan （LNT） Methods （1）Observing the phagocytosis of macrophages in murine cavity induced by LNT，（2） Measuring the tumor necrosis factor （TNF）of L615 leukeamia mice after being injected by LNT,（3） Apoptosis mediated by TNF in macrophages induced by LNT. was detected with Hochest 33258 fluorochrome staining and DNA electrophoresis.Results The ratio and index of phagocytosis of macrophages in murine cavity induced by lentinan （LNT） were increased significantly.Meanwhile content of tumor necrosis factor（TNF） in supernatant of macrophages splenic cells and thymocytes induced by LNT were higher than the control.The cytoxic activation of macrophage supernatant induced by LNT on L615 leukeamia cells rose to 66.2% from the control （11.4%） and expressed a dose dependeant manner when L615 leukeamia cells were treated by the macrophage supernatant induced by LNT DNA,the apoptotic cells were found.Conclusion The macrophages activated by LNT shows high cytotoxic activation on tumor cells, and one of its mechanisms of anti-tumor cells is the apoptosis mediated by TNF in macrophages induced by LNT. 【Key words】 lentinan；macrophage；TNF;apptosis 现代研究证明，香菇多糖（LNT）可调节人体内有免疫功能的T细胞活性，可降低甲基胆蒽诱发肿瘤的能力。香菇对癌细胞有强烈的抑制作用。从细胞凋亡（Apoptosis）的角度探讨抗肿瘤途径是目前医学研究的热点。细胞凋亡机制比较复杂，而且不单纯是一个生理过程，它与很多免疫的发生发展有着重要的关系［1,2］。利用某些手段诱导肿瘤细胞凋亡进而发挥抗肿瘤目的是本课题出发点。我们利用 LNT诱导活化巨噬细胞进而介导肿瘤细胞凋亡，起到抗肿瘤效果，更符合生物自身防御原则，达到扶正固本的作用。 1 材料与方法 1.1 巨噬细胞功能测定 615纯系小鼠（4～6周龄，中国医科院血研所动物中心）静脉注射LNT［Lentinus edodes（Berk） Sing干燥子实体中提取，福建梅峰制药厂］66.7mg/㎏体重，qd× 7d