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目的:探讨片仔癀诱导骨肉瘤U2OS细胞凋亡与线粒体跨膜电位(Δψm)的关系。方法:采用不同浓度的片仔癀(0.8、1.2、1.6mg/mL)作用U2OS细胞后,Hoechst 33258染色镜下观察、Annexin V-FITC/PI双染流式细胞仪检测细胞凋亡情况;JC-1染色流式细胞仪检测线粒体跨膜电位;分光光度法检测Caspase-9、Caspase-3活性;Western blot检测Bcl-2、Bax蛋白的表达。结果:随剂量的增加,片仔癀可明显诱导骨肉瘤U2OS细胞凋亡、降低细胞线粒体膜电位,增加Caspase-9、Caspase-3活性,各干预组与阴性对照组比较均有明显差异(P<0.05,P<0.01),Western blot检测结果表明Bcl-2蛋白表达下降,而Bax蛋白表达上调。结论:片仔癀可降低细胞线粒体跨膜电位,改变线粒体通透性,促使骨肉瘤细胞凋亡。
Objective: To investigate the relationship between the apoptosis of U2OS cells and mitochondrial transmembrane potential (Δψm) induced by Pien Tze Huang. Methods: U2OS cells were treated with different concentrations of Pien Tze Huang (0.8,1.2,1.6mg / mL). Hoechst 33258 staining and Annexin V-FITC / PI double staining were used to detect apoptosis. JC-1 The transmembrane potential of mitochondria was detected by flow cytometry. The activities of Caspase-9 and Caspase-3 were detected by spectrophotometry. The expressions of Bcl-2 and Bax protein were detected by Western blot. Results: With the increase of dose, Pien Tze Huang significantly induced the apoptosis of osteosarcoma U2OS cells, decreased the mitochondrial membrane potential and increased the activity of Caspase-9 and Caspase-3. There was significant difference between each intervention group and the negative control group (P <0.05 , P <0.01). The results of Western blot showed that the expression of Bcl-2 protein was down-regulated and the expression of Bax protein was up-regulated. Conclusion: Pien Tze Huang can reduce mitochondrial transmembrane potential, change mitochondrial permeability and promote osteosarcoma cell apoptosis.