姜黄素诱导永生化人HaCaT细胞凋亡

目的 探讨姜黄素对人永生化上皮细胞凋亡的诱导作用.方法 应用细胞计数、流式细胞术、琼脂糖凝胶电泳、Hoechst33258染色、苏木素-伊红(HE)染色和透射电镜检测经姜黄素诱导处理后人永生化上皮HaCaT细胞的凋亡.结果经7.5mg/L姜黄素诱导处理后,人永生化上皮HaCaT细胞的增殖活动受到明显的抑制,细胞生长抑制率达83.03%;细胞周期检测出现亚二倍体(亚G1期)细胞峰值,细胞凋亡率达13.1%,并发生G2/M期阻滞;琼脂糖凝胶电泳出现细胞凋亡典型的DNA"梯状"条带;细胞核经Hoechst33258染色出现浓染致密的固缩形态和颗粒状荧光;光镜和电镜观察结果显示,姜黄素处理组细胞体积缩小,细胞核固缩,染色质凝聚,线粒体肿胀,有凋亡小体形成等显著的凋亡特征.结论姜黄素对人永生化上皮HaCaT细胞凋亡有显著的诱导作用,从而为进一步研究表皮细胞衰老、凋亡机理提供了重要基础和研究依据.

Curcumin induces apoptosis of immortalized human epithelial cell line HaCaT

Objective To study the apoptosis effect of curcumin(Cur) on the immortalized human epithelial cell line HaCaT. Methods The apoptosis effect were detected cell by cell count, flow cytometry analysis, DNA gel electrophoresis, Hoechst33258 staining,HE staining, and electron microscopy. Results After treated with 7.5mg/L curcumin the proliferation of HaCaT cells were inhibited, and the inhibitory rate was 83.03%. The results of flow cytometry analysis showed that curcumin could induced the emergence of the phase of apoptosis, and the rate was 13.1%,the cell cycle were arrested in GSUB>2/SUB>/M phase. Agarose gel electrophoresis revealed that cell DNA fragment exhibited characteristic “DNA ladder”. Cell nucleus concentrated and appeared granular fluorescence by Hoechst33258 staining. Light microscope and electron microscope showed that the cells treated with curcumin shrinked, cell nucleus concerntrated.chromatin agglutinated, mitochondria swelled, and apoptosis body formed. Conclusion This study suggests that curcumin could induce apoptosis of immortalized human epithelial cell line HaCaT effectively, and provided an important foundation for further studying the consenescence and apoptosis mechanisms of the epid