露蜂房蛋白体外诱导K562细胞凋亡的作用及其机制

Apoptosis of K562 cells induced by proteins extracted from Nidus Vespae in vitro and its mechanism

目的探讨露蜂房蛋白(NVP)体外诱导人红白血病细胞系K562细胞凋亡的作用及其机制。方法将露蜂房蛋白NVP1、NVP2、NVP3及NVP4分别作用于K562细胞,采用细胞培养、Hoechst 33258荧光染色技术观察K562细胞形态变化;采用免疫组织化学方法检测K562细胞天冬氨酸特异性半胱氨酸蛋白酶-3(cas-pase-3)表达。结果NVP1~NVP4作用后K562细胞生长缓慢,数量减少,胞质内颗粒增多,并出现大量空泡,细胞碎片逐渐增多;Hoechst33258荧光染色显著增强,染色质呈致密浓染的块状或粗颗粒状荧光。K562细胞cas-pase-3蛋白表达明显增加,与空白对照组相比有显著性差异(P<0.05)。结论NVP能抑制K562细胞增殖并诱导其凋亡;其作用机制可能是使caspase-3蛋白表达升高和活化。

[Objective] To explore proteins from Nidus Vespae （NVP） inducing apoptosis of K562 cells in vitro and its mechanism. [Methods] Protein ingredients were extracted from NVP by various means including centrifuge,gel chromatography and dialysis. K562 cells were cultured with NVP1-4. Morphological changes of apoptotic cells were observed via an inverted phase contrast microscope and by Hoechst 33258 stain via a fluorescent microscope （FM）. Expressions of caspase-3 was examined by immunohistochemistry. [Results] K562 cells growed more slowly and were decreased. Particles in cytoplasm were increased and there were many cacuoles and fragments cytoplasm. Fluorescence staining enhanced significantly, karyotin showed dense lumps or thick particles. Caspase-3 expression was increased significantly. Compared with blank control group NVP groups all had significant differences （P^0.05）. [Conclusion] NVP can markedly inhibit the proliferation of K562 cells. The mechanism may be related to upregulation and activation of caspase-3.