Zhang Lu-min, Lv Xue-wei, Shao Lin-xiang, Ma Yan-fang, Cheng Wen-zhao, Gao Hai-tao
Zhong Yao Cai. 2013 Dec;36(12):1988-92.
To investigate the effects of Artemisia lavandulaefolia essential oil on apoptosis and necrosis of HeLa cells.
Cell viability was assayed using MTT method. The morphological and structure alterations in HeLa cells were observed by microscopy. Furthermore, cell apoptosis was measured by DNA Ladder and flow cytometry. DNA damage was measured by comet assay, and the protein expression was examined by Western blot analysis.
MTT assay displayed essential oil from Artemisia lavandulaefolia could inhibit the proliferation of HeLa cells in a dose-dependent manner. After treated with essential oil of Artemisia lavadulaefolia for 24 h, HeLa cells in 100 and 200 microg/mL experiment groups exhibited the typical morphology changes of undergoing apoptosis, such as cell shrinkage and nucleus chromatin condensed. However, the cells in the 400 microg/mL group showed the necrotic morphology changes including cytomembrane rupture and cytoplasm spillover. In addition, DNA Ladder could be demonstrated by DNA electrophoresis in each experiment group. Apoptosis peak was also evident in flow cytometry in each experiment group. After treating the HeLa cells with essential oil of Artemisia lavadulaefolia for 6 h, comet tail was detected by comet assay. Moreover, western blotting analysis showed that caspase-3 was activated and the cleavage of PARP was inactivated.
Essential oil from Artemisia lavadulaefolia can inhibit the proliferation of HeLa cells in vitro. Low concentration of essential oil from Artemisia lavadulaefolia can induce apoptosis, whereas high concentration of the compounds result in necrosis of HeLa cells. And,the mechanism may be related to the caspase-3-mediated-PARP apoptotic signal pathway.
探讨薰衣草叶艾精油对人宫颈癌HeLa细胞凋亡和坏死的影响。
采用MTT法检测细胞活力。通过显微镜观察HeLa细胞的形态和结构变化。此外,采用DNA Ladder法和流式细胞术检测细胞凋亡。采用彗星试验检测DNA损伤,通过蛋白质印迹分析检测蛋白表达。
MTT试验显示薰衣草叶艾精油能以剂量依赖性方式抑制HeLa细胞的增殖。用薰衣草叶艾精油处理24 h后,100和200 μg/mL实验组的HeLa细胞呈现典型的凋亡形态变化,如细胞收缩和细胞核染色质浓缩。然而,400 μg/mL组的细胞表现出坏死形态变化,包括细胞膜破裂和细胞质外溢。此外,各实验组的DNA电泳均能显示出DNA Ladder。各实验组的流式细胞术中也可见凋亡峰。用薰衣草叶艾精油处理HeLa细胞6 h后,彗星试验检测到彗星尾。此外,蛋白质印迹分析显示caspase-3被激活,PARP的裂解被抑制。
薰衣草叶艾精油可在体外抑制HeLa细胞的增殖。低浓度的薰衣草叶艾精油可诱导凋亡,而高浓度的该化合物则导致HeLa细胞坏死。其机制可能与caspase-3介导的PARP凋亡信号通路有关。