露蜂房纯化蛋白对白血病细胞形态学影响的研究

Effect of the purified protein from Nidus Vespae on the morphology of leukemic cell

目的观察中药露蜂房纯化蛋白Ⅱ(nidus vespae protein-Ⅱ,NVP-Ⅱ)对人早幼粒白血病细胞株HL-60和急性髓系白血病(AML)患者骨髓单个核细胞(BMMNC)形态学的影响。方法采用体外细胞培养,加入不同浓度NVP-Ⅱ作用于HL-60细胞和AML患者BMMNC,培养72 h后,生长曲线法测定NVP-Ⅱ对白血病细胞增殖的影响,倒置显微镜下观察细胞生长状况,并收集细胞常规制备超薄切片,透射电子显微镜下观察HL-60细胞和AML患者BMMNC的超微结构。结果光镜下NVP-Ⅱ处理组白血病细胞数量明显减少,并出现细胞碎片;透射电镜下,NVP-Ⅱ处理组白血病细胞出现核固缩,染色质浓缩边集,呈新月状、块状或碎裂为质膜包绕的团块,形成凋亡小体;线粒体基质深染,出现空泡样变。结论NVP-Ⅱ有明显诱导白血病细胞凋亡的作用,并呈现典型的细胞凋亡形态学改变。

Objective To observe the effect of the purificated protein from Nidus Vespae（ NVP-Ⅱ） on the morphologic changes of HL-60 cells and bone marrow mononuelear cells （ BMMNC ） of acute myeloid leukemia （ AML ） patient. Methods Treated cultured HL-60 cells with various concentration of NVP-Ⅱ for 72 hours in vitro, using growth curve to detect cell proliferation and using inverted microscope to observe the growth of leukemia cells. Transmission electronic microscope was used to observe the uhrastructural changs of the ceils. Results The number of leukemia cells obviously decreased and cell fragments appeared under light microscope. Under transmission electronic microscope, typical morphological changes of apoptosis were observed among HL-60 cells and BMMNC of AML patient in NVP-Ⅱ treated groups, such as nuclear chromatin condensation and marginized to be crescent and formed apoptotic body; hyperchomatin and vacuolar degeneration of mitochondria appeared. Conclusion NVP-Ⅱ can induce typical morphologic changes of apoptosis of HL-60 cells in vitro.