氨肽酶抑制剂Bestatin通过激活半胱天冬酶3诱导HL-60细胞凋亡

目的 研究半胱天冬酶3在国产氨肽酶N抑制剂Bestatin(商品名百士欣，BS）诱导人白血病细胞凋亡过程中的变化及其意义。方法 用光学显微镜观察细胞形态结构的改变，通过DNA片段原位末端标记法及流式细胞术（FCM）检测细胞凋亡。用发色底物法检测细胞半胱天冬酶3活性。Rhodamin(Rh)123染色后，采用FCM检测细胞线粒体跨膜电位。结果 典型的细胞形态改变、DNA片段化、DNA末端原位标记及FCM结果，均证实BS能诱导HL－60细胞凋亡。凋亡率与药物剂量和作用时间呈依赖关系。BS诱导细胞凋亡过程中，半胱天冬酶3活性明显升高，而AC0DEVD－CHO能抑制BS诱导的细胞凋亡。经BS处理的HL－60细胞出现线粒体跨膜电位（ΔΨm)下降。结论 BS通过激活半胱天冬酶3而诱导白血病细胞凋亡。

Amiopeptidase inhihitor Bestatin induces HL-60 cell apoptosis through activating caspase 3

OBJECTIVE: To study the variation and significance of caspase 3 activity in the process of amino-peptidase inhibitor--bestatin (BS) inducing human leukemic cell apoptosis. METHODS: Cell apoptosis was evaluated by light microscopy, TUNEL labeling and flow cytometry (FCM). Caspase 3 activity was detected by colorimetry. The mitochondrial transmembrane potentials (DeltaPsi(m)) were detected by Rhodamine123 staining. RESULTS: The apoptotic morphology, apoptotic peak on FCM and positive Annexin V(FITC) on cell membrane showed that BS could induce HL-60 cell apoptosis in a dose- and time-dependent manner. Caspase 3 activity was significantly higher in the apoptotic cells than in control cells. The apoptosis induced by BS was inhibited by AC-DEVD-CHO. The DeltaPsi(m) of cells treated with BS declined. CONCLUSION: BS induces apoptosis of human acute leukemic cells through activation of caspase 3.