Homeopathic mother tincture of Phytolacca decandra induces apoptosis in skin melanoma cells by activating caspase-mediated signaling via reactive oxygen species elevation
Homeopathic mother tincture of Phytolacca decandra induces apoptosis in skin melanoma cells by activating caspase-mediated signaling via reactive oxygen species elevation
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DOI：
10.3736/jintegrmed2013014
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发表时间：
2013-03-01
影响因子：
4.8
通讯作者：
Khuda-Bukhsh, Anisur Rahman
中科院分区：
文献类型：
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作者：
Ghosh, Samrat;Bishayee, Kausik;Khuda-Bukhsh, Anisur Rahman
目的：针对皮肤黑色素瘤的预防措施（如化疗）是有用的，但存在慢性副作用和耐药性。商陆 (PD) 的乙醇提取物，用于顺势疗法治疗各种疾病，如慢性风湿病、定期结膜炎、牛皮癣，并在某些皮肤病中测试其可能的抗癌潜力。方法：通过对正常人和正常人进行 3-(4,5-二甲基噻唑-2-y1)-2,5-二苯基四唑溴化物测定来测试该药物的细胞毒性。 （外周血单核细胞）和A375细胞。对 4',6-二脒基-2-苯基吲哚二盐酸盐染色的细胞进行荧光显微镜研究以进行 DNA 片段化分析，并记录细胞形态的变化（如果有）。进行乳酸脱氢酶活性测定以评估细胞凋亡和坏死的百分比。活性氧 (ROS) 积累（如果有）以及凋亡基因的表达研究也进行了评估，以查明凋亡的实际事件。 结果：结果显示，PD 给药导致 A375 细胞增殖显着减少，但对外周血单核细胞没有表现出太大的细胞毒性。 PD 处理的细胞中，ROS 的产生和 DNA 损伤（使癌细胞易于凋亡）增强。这些结果得到了不同细胞和核蛋白表达的分析数据的充分支持，例如，通过下调 Akt 和 Bcl-2，上调 p53、Bax 和 caspase 3，以及 A375 细胞中细胞凋亡导致的细胞死亡数量增加。结论：总体结果表明 PD 通过激活 caspase 介导的信号传导和 ROS 生成对 A375 细胞具有抗癌潜力。
OBJECTIVE: Preventive measures against skin melanoma like chemotherapy are useful but suffer from chronic side effects and drug resistance. Ethanolic extract of Phytolacca decandra (PD), used in homeopathy for the treatment of various ailments like chronic rheumatism, regular conjunctivitis, psoriasis, and in some skin diseases was tested for its possible anticancer potential.METHODS: Cytotoxicity of the drug was tested by conducting 3-(4,5-dimethylthiazol-2-y1)-2,5diphenyltetrazoliurn bromide assay on both normal (peripheral blood mononuclear cells) and A375 cells. Fluorescence microscopic study of 4',6-diamidino-2-phenylindole dihydrochloride-stained cells was conducted for DNA fragmentation assay, and changes in cellular morphology, if any, were also recorded. Lactate dehydrogenase activity assay was done to evaluate the percentages of apoptosis and necrosis. Reactive oxygen species (ROS) accumulation, if any, and expression study of apoptotic genes also were evaluated to pin-point the actual events of apoptosis.RESULTS: Results showed that PD administration caused a remarkable reduction in proliferation of A375 cells, without showing much cytotoxicity on peripheral blood mononuclear cells. Generation of ROS and DNA damage, which made the cancer cells prone to apoptosis, were found to be enhanced in PD-treated cells. These results were duly supported by the analytical data on expression of different cellular and nuclear proteins, as for example, by down regulation of Akt and BcI-2, up-regulation of p53, Bax and caspase 3, and an increase in number of cell deaths by apoptosis in A375 cells.CONCLUSION: Overall results demonstrate anticancer potentials of PD on A375 cells through activation of caspase-mediated signaling and ROS generation.