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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。
Objective To investigate the induction of curcumin on human immortalized epithelial cells. Methods The apoptosis of immortalized human HaCaT cells induced by curcumin was detected by cell counting, flow cytometry, agarose gel electrophoresis, Hoechst33258 staining, hematoxylin and eosin (HE) staining and transmission electron microscopy. Results After induced by 7.5 mg / L curcumin, the proliferation activity of human immortalized HaCaT cells was significantly inhibited and the rate of cell growth inhibition was 83.03%. The peak of sub-diploid (sub-G1 phase) cells was detected in cell cycle, Apoptosis rate reached 13.1%, and G2 / M phase arrest occurred. A typical DNA “ladder” banding of apoptosis appeared on agarose gel electrophoresis. The nucleus was dense and densely packed by Hoechst33258 staining And granular fluorescence. The results of light microscope and electron microscope showed that curcumin treated cells showed obvious apoptotic features such as reduced cell volume, nuclear condensation, chromatin condensation, mitochondria swelling and formation of apoptotic bodies. Conclusion Curcumin induces the apoptosis of human immortalized epithelial HaCaT cells, which provides an important basis and basis for further study on the mechanism of epidermal cell aging and apoptosis.