露蜂房纯化蛋白对急性髓系白血病患者骨髓细胞的影响

目的研究中药露蜂房纯化蛋白(nidus vespae protein-Ⅱ,NVP-Ⅱ)对急性髓系白血病(AML)患者骨髓单个核细胞(BMMNC)的作用并探讨其机制。方法采用NVP-Ⅱ在体外作用于AML患者BMMNC,经光镜、电镜、Hochest33258染色法、TdT缺口末端标记(TUNEL)技术等方法检测NVP-Ⅱ对AML患者BMMNC的影响。结果不同浓度NVP-Ⅱ对AML患者BMMNC有不同程度的生长抑制和诱导凋亡作用。①光镜下观察到,与生理盐水对照组比较,NVP-Ⅱ处理组AML患者BMMNC数量明显减少,胞质内颗粒增多,并有较多的细胞碎片。②透射电镜下发现,NVP-Ⅱ处理组AML患者BMMNC细胞核染色质浓缩、碎裂,靠核膜边集呈块状,并形成凋亡小体;线粒体基质深染,呈现空泡样变及髓样变。③Hochest33258染色法显示,NVP-Ⅱ处理组AML患者BMMNC核染色质高度凝聚,致密浓染,或裂解呈碎块状,边缘光滑清晰。④TUNEL检测见阳性细胞核深染,NVP-Ⅱ处理组与生理盐水处理组比较,凋亡细胞明显增多(P<0.05)。结论NVP-Ⅱ可抑制AML患者BMMNC生长并诱导其凋亡。

Effect of purified protein from nidus vespae on bone marrow mononuclear cells of acute myeloid leukemia

Aim To study the effect of purified protein extracted from Nidus Vespae(NVP-Ⅱ) on bone marrow mononuclear cells (BMMNC) of the patients with acute myeloid leukemia (AML). Method BMMNC of AML patients were treated with NVP-Ⅱ in vitro. Light microscopy, electron microscopy, Hochest33258 dying method and TUNEL were used to investigate the effect. Results (1)Compared with saline control group, the number of BMMNC of AML patients in NVP-Ⅱ group decreased obviously; more granules appeared in cytoplasm; many leukemia cells broke into several fragments. (2) By means of transmission electronic microscopy, it was observed that nuclear chromatin condensed and marginalized to be nubby and formed apoptotic body; organelle such as ribosome, rough endoplasm reticulum and mitochondrion clustered; mitochondrial basal substance was stained deeply , and vacuolar and myeloid degeneration appeared in BMMNC of AML patients in NVP-Ⅱ group.(3) Hochest33258 dying showed that the morphological changes of BMMNC of AML patients in NVP-Ⅱgroup ,such as compaction, condensation and segregation of the nuclear chromatin appeared.(4) Positive cells of hyperchromism were observed with TUNEL. Apoptotic cells increased significantly in NVP-Ⅱ group, comparing with saline control group(P<0.05). Conclusion NVP-Ⅱ can inhibit the proliferation and induce the apoptosis of BMMNC of AML patients in vitro.