【摘要】 目的　研究冬凌草甲素诱导人黑色素瘤 A375 - S2细胞凋亡的作用机制。方法　 MTT法检测冬凌草甲素对 A375 - S2细胞生长的影响及各种 Caspase抑制剂、佛波酯 (PMA)、PKC抑制剂 H- 7对冬凌草甲素作用的影响 ;DNA凝胶电泳法检测细胞凋亡 ;L DH法测定乳酸脱氢酶 (L DH)活力。结果　冬凌草甲素对 A375 - S2细胞生长有显著抑制作用 ,并能显著诱导 A375 - S2细胞发生凋亡 ,其作用呈明显的量效关系和时间依赖性。形态学观察可见凋亡小体的形成 ,琼脂糖凝胶电泳可见凋亡典型的 DNA梯带 ;Caspase家族抑制剂 ,Caspase- 9抑制剂能显著抑制冬凌草甲素诱导 A375 - S2细胞的凋亡。大剂量 PMA(4 0 0 ng/m L)显著抑制 34.3μmol/L 冬凌草甲素诱导 A375 - S2细胞凋亡 ,而小剂量 PMA(10 ng/m L)与冬凌草甲素有协同杀死细胞的作用 ,且 PKC抑制剂 H- 7也可下调这种凋亡作用。 Caspase- 3的抑制剂可抑制 Caspase- 3的活力升高。结论　适宜浓度的冬凌草甲素 (34.3μm ol/L )诱导A375 - S2细胞凋亡 ,这种作用是通过线粒体调节 Caspase的激活来实现的。更多还原

【Abstract】 Object To study the mechanisms of oridonin-induced A375-S2 cell apoptosis. Methods MTT assay, morphological observation, agarose gel electrophoresis, LDH release and Caspase inhibitors, PMA, PKC were used. Results Oridonin had significant apoptotic effect on A375-S2 cells in a dose- and a time-dependent manners. The marked morphological changes including condensed chromatin, nuclear fragmentation and apoptotic bodies; and DNA ladder in agarose gel were observed. The activity of Caspase-3 was increased after treatment with oridonin about 6 times as much as the control value, since high dose PMA inhibited PKC activity, low dose PMA activated PKC. In this study, 400 ng/mL PMA blocked 34.3 μmol/L oridonin-induced A375-S2 cell apoptosis and PKC inhibitor H-7 down-regulated the apoptotic effect, however, low dose PMA (10 ng/mL) more sensitized A375-S2 cell to oridonin. Conclusion Oridonin (34.3 μmol/L) can induce A375-S2 cell apoptosis via mitochondria-regulated Caspase pathway activation, PKC activation is involved in this mechanism.更多还原