Centella asiatica Fraction-3 Suppresses the Nuclear Factor Erythroid 2-Related Factor 2 Anti-Oxidant Pathway and Enhances Reactive Oxygen Species-Mediated Cell Death in Cancerous Lung A549 Cells
Centella asiatica Fraction-3 Suppresses the Nuclear Factor Erythroid 2-Related Factor 2 Anti-Oxidant Pathway and Enhances Reactive Oxygen Species-Mediated Cell Death in Cancerous Lung A549 Cells
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DOI：
10.1089/jmf.2017.0005
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发表时间：
2017-10-01
影响因子：
2.4
通讯作者：
Chuturgoon, Anil Amichund
中科院分区：
文献类型：
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作者：
Naidoo, Dhaneshree Bestinee;Phulukdaree, Alisa;Chuturgoon, Anil Amichund
积雪草是一种热带药用植物，通常用于传统医学。药用价值C.包括抗氧化、抗炎和抗癌活性。我们研究了C的半纯化部分的抗氧化和抗增殖/细胞毒性作用。积雪草叶乙醇提取物（C3）在癌性肺A549细胞中的作用。通过硅胶柱分离得到C3，并通过薄层色谱和气相色谱质谱鉴定。评价C3在A549细胞中的细胞毒性（细胞活力试验-WST-1; 24 h; [0.23 mg/mL]），以测定抑制浓度（IC 50）。评价细胞内活性氧（IROS）、线粒体膜电位（流式细胞术）、丙二醛（MDA）、乳酸脱氢酶（LDH）（分光光度法）、谷胱甘肽（GSH）、氧化型谷胱甘肽（GSSG）、三磷酸腺苷水平、半胱天冬酶活性（光度法）和DNA损伤（彗星试验）。蛋白质表达（Nrf-2、p53、Bax、Bcl-2和HSP-70）和基因表达（Nrf-2、GPx、SOD、CAT、c-myc和OGG-1）分别通过蛋白质印迹和定量聚合酶链反应（qPCR）进行定量。C3剂量依赖性地降低A549细胞活力。C3的IC 50增加了MDA、IROS、线粒体去极化、LDH、caspase（-8，-9，-3/7）活性、DNA损伤、GSH水平、Nrf-2蛋白表达、HSP-70蛋白表达和OGG-1基因表达（P < .05）。C3可降低GSSG水平、抗氧化剂（Nrf-2、GPx、SOD）基因表达、p53、Bax和Bcl-2蛋白表达（P <0.02）。C3降低了A549细胞中抗氧化基因的表达并诱导了抗增殖/细胞毒性作用。
Centella asiatica is a tropical medicinal plant that is commonly used in traditional medicine. Medicinal properties of C. asiatica include anti-oxidant, anti-inflammatory, and anti-cancer activity. We investigated the anti-oxidant and anti-proliferative/cytotoxic effects of a semi-purified fraction of C. asiatica ethanolic leaf extract (C3) in cancerous lung A549 cells. C3 was obtained by silica column fractionation and identified by using thin-layer chromatography and gas chromatography mass spectrometry. Cytotoxicity of C3 in A549 cells was evaluated (cell viability assay-WST-1; 24 h; [0.23 mg/mL]) to determine an inhibitory concentration (IC50). Intracellular reactive oxygen species (IROS), mitochondrial membrane potential (flow cytometry), malondialdehyde (MDA), lactate dehydrogenase (LDH) (spectrophotometry), glutathione (GSH), oxidised glutathione (GSSG), adenosine triphosphate levels, caspase activity (luminometry), and DNA damage (comet assay) were evaluated. Protein expression (Nrf-2, p53, Bax, Bcl-2, and HSP-70) and gene expression (Nrf-2, GPx, SOD, CAT, c-myc, and OGG-1) were quantified by western blotting and quantitative polymerase chain reaction (qPCR), respectively. C3 dose dependently decreased A549 cell viability. The IC50 of C3 increased MDA, IROS, mitochondrial depolarization, LDH, caspase (-8, -9, -3/7) activity, DNA damage, GSH levels, Nrf-2 protein expression, HSP-70 protein expression, and OGG-1 gene expression (P < .05). GSSG levels, anti-oxidant (Nrf-2, GPx, SOD) gene expression, p53, Bax, and Bcl-2 protein expression were decreased by C3 (P < .02). C3 diminished the anti-oxidant gene expression and induced anti-proliferative/cytotoxic effects in A549 cells.