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 ...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-yl）-2,5- diphenyltetrazolium 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 Bcl-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.展开更多
目的:东莨菪亭(7-羟基-6甲氧基香豆素,化学式C10H8O4,简称HMC)是从常绿钩吻(Gelsemium sem pervirens)中提取的一种天然香豆素类化合物,被认为具有抗癌活性。本研究将HMC制成聚乳酸聚乙醇酸共聚物(polylactic-co-glycolic acid,PLGA)纳...目的:东莨菪亭(7-羟基-6甲氧基香豆素,化学式C10H8O4,简称HMC)是从常绿钩吻(Gelsemium sem pervirens)中提取的一种天然香豆素类化合物,被认为具有抗癌活性。本研究将HMC制成聚乳酸聚乙醇酸共聚物(polylactic-co-glycolic acid,PLGA)纳米颗粒胶囊,通过多项指标观察其与非纳米HMC相比,在被细胞摄取、生物利用度及对细胞凋亡的影响(抗癌活性)等方面是否有所提高。方法:人黑色素瘤A375细胞用来检测HMC及纳米HMC颗粒(nano-7-hydroxy-6-methoxy coumarin,NHMC)的被细胞摄取率及抗癌活性;通过动态光散射确定NHMC的颗粒大小、多分散性指数及电动电位;扫描电子显微镜及原子力显微镜检测纳米颗粒的表面形态。结果:HMC制成纳米颗粒胶囊的包封率超过85%。NHMC颗粒的平均直径小于110nm,其多分散性系数为0.237,提高了其被细胞摄取率及生物活性。NHMC被细胞摄取的时间(15min)明显少于非纳米HMC(30min)。测定细胞内某些信号分子mRNA的表达,表明NHMC作用细胞后能够下调细胞周期素D1(cyclin-D1)、增殖细胞核抗原(PCNA)、存活素(survivin)及信号转导和转录活化因子3(Stat-3)的表达并上调p53及半胱天冬酶3(caspase-3)的表达。与非纳米HMC相比,NHMC能够更快地被细胞摄取并引起更多的肿瘤细胞凋亡。NHMC对正常皮肤细胞无明显细胞毒性。结论:使用生物可降解材料PLGA制成的小分子纳米颗粒NHMC能够更快速地被细胞摄取并引起更多的肿瘤细胞凋亡,而对正常皮肤细胞无明显细胞毒性。提取天然药用植物中的有效成分制成纳米颗粒胶囊有望成为更好地发挥其药效的途径。展开更多
OBJECTIVE： To examine if the ultra-highly diluted homeopathic remedy Thuja 30C can ameliorate benzo（a）pyrene （BaP）-induced DNA damage, stress and viability of perfused lung cells of Swiss albino mice in vitro. ME...OBJECTIVE： To examine if the ultra-highly diluted homeopathic remedy Thuja 30C can ameliorate benzo（a）pyrene （BaP）-induced DNA damage, stress and viability of perfused lung cells of Swiss albino mice in vitro. METHODS： Perfused normal lung cells from mice were cultured in 5% Roswell Park Memorial Institute medium and exposed to BaP, a potent carcinogen, at the half maximal inhibitory concentration dose （2.2 μmol/L） for 24 h. Thereafter, the intoxicated cells were either treated with Thuja 30C （used against tumor or cancer） or its vehicle media, succussed alcohol 30C. Relevant parameters of study involving reactive oxygen species （ROS） accumulation, total glutathione （GSH） content, and generations of heat shock protein （hsp）-90 were measured; the cell viability and other test parameters were measured after treatment with either Thuja 30C or its vehicle media. Circular dichroism spectroscopy was performed to examine if Thuja 30C directly interacted with calf thymus DNA as target. For ascertaining if DNA damaged by BaP could be partially repaired and restituted by the remedy, 4＇,6-diamidino-2-phenylindole staining was performed. RESULTS： Thuja 30C increased cell viability of BaP-intoxicated cells significantly, as compared to drug-untreated or drug-vehicle control. A minimal dose of Thuja 30C significantly inhibited BaP-induced stress level, by down-regulating ROS and hsp-90, and increasing GSH content. Thuja 30C itself had no DNA-damaging effect, and no direct drug-DNA interaction. However, it showed quite striking ability to repair DNA damage caused by BaP. CONCLUSION： Thuja 30C ameliorates BaP-induced toxicity, stress and DNA damage in perfused lung cells of mice and it apparently has no effect on normal lung cells.展开更多
基金financially supported by a grant sanctioned to Prof. A.R. Khuda-Bukhsh, Department of Zoology,University of Kalyani, by Boiron Laboratories, Lyon, France
文摘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-yl）-2,5- diphenyltetrazolium 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 Bcl-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.
基金Boiron Laboratory supported this work financially by a research grant sanctioned to Prof . A.R. Khuda-Bukhsh.
文摘目的:东莨菪亭(7-羟基-6甲氧基香豆素,化学式C10H8O4,简称HMC)是从常绿钩吻(Gelsemium sem pervirens)中提取的一种天然香豆素类化合物,被认为具有抗癌活性。本研究将HMC制成聚乳酸聚乙醇酸共聚物(polylactic-co-glycolic acid,PLGA)纳米颗粒胶囊,通过多项指标观察其与非纳米HMC相比,在被细胞摄取、生物利用度及对细胞凋亡的影响(抗癌活性)等方面是否有所提高。方法:人黑色素瘤A375细胞用来检测HMC及纳米HMC颗粒(nano-7-hydroxy-6-methoxy coumarin,NHMC)的被细胞摄取率及抗癌活性;通过动态光散射确定NHMC的颗粒大小、多分散性指数及电动电位;扫描电子显微镜及原子力显微镜检测纳米颗粒的表面形态。结果:HMC制成纳米颗粒胶囊的包封率超过85%。NHMC颗粒的平均直径小于110nm,其多分散性系数为0.237,提高了其被细胞摄取率及生物活性。NHMC被细胞摄取的时间(15min)明显少于非纳米HMC(30min)。测定细胞内某些信号分子mRNA的表达,表明NHMC作用细胞后能够下调细胞周期素D1(cyclin-D1)、增殖细胞核抗原(PCNA)、存活素(survivin)及信号转导和转录活化因子3(Stat-3)的表达并上调p53及半胱天冬酶3(caspase-3)的表达。与非纳米HMC相比,NHMC能够更快地被细胞摄取并引起更多的肿瘤细胞凋亡。NHMC对正常皮肤细胞无明显细胞毒性。结论:使用生物可降解材料PLGA制成的小分子纳米颗粒NHMC能够更快速地被细胞摄取并引起更多的肿瘤细胞凋亡,而对正常皮肤细胞无明显细胞毒性。提取天然药用植物中的有效成分制成纳米颗粒胶囊有望成为更好地发挥其药效的途径。
基金supported by a grant sanctioned to Prof.A.R.Khuda-Bukhsh,Depart ment of Zoology,University of Kalyani,Kalyani-741235,India by Boiron Laboratory,Lyon,France
基金financially supported by a grant sanctioned to Prof.A.R.Khuda-Bukhsh,Department of Zoology, University of Kalyani,India,by Boiron Laboratories,Lyon, France
文摘OBJECTIVE： To examine if the ultra-highly diluted homeopathic remedy Thuja 30C can ameliorate benzo（a）pyrene （BaP）-induced DNA damage, stress and viability of perfused lung cells of Swiss albino mice in vitro. METHODS： Perfused normal lung cells from mice were cultured in 5% Roswell Park Memorial Institute medium and exposed to BaP, a potent carcinogen, at the half maximal inhibitory concentration dose （2.2 μmol/L） for 24 h. Thereafter, the intoxicated cells were either treated with Thuja 30C （used against tumor or cancer） or its vehicle media, succussed alcohol 30C. Relevant parameters of study involving reactive oxygen species （ROS） accumulation, total glutathione （GSH） content, and generations of heat shock protein （hsp）-90 were measured; the cell viability and other test parameters were measured after treatment with either Thuja 30C or its vehicle media. Circular dichroism spectroscopy was performed to examine if Thuja 30C directly interacted with calf thymus DNA as target. For ascertaining if DNA damaged by BaP could be partially repaired and restituted by the remedy, 4＇,6-diamidino-2-phenylindole staining was performed. RESULTS： Thuja 30C increased cell viability of BaP-intoxicated cells significantly, as compared to drug-untreated or drug-vehicle control. A minimal dose of Thuja 30C significantly inhibited BaP-induced stress level, by down-regulating ROS and hsp-90, and increasing GSH content. Thuja 30C itself had no DNA-damaging effect, and no direct drug-DNA interaction. However, it showed quite striking ability to repair DNA damage caused by BaP. CONCLUSION： Thuja 30C ameliorates BaP-induced toxicity, stress and DNA damage in perfused lung cells of mice and it apparently has no effect on normal lung cells.
