Isolation and characterization of gallic acid and methyl gallate from the seed coats of Givotia rottleriformis Griff. and their anti-proliferative effect on human epidermoid carcinoma A431 cells.
Isolation and characterization of gallic acid and methyl gallate from the seed coats of Givotia rottleriformis Griff. and their anti-proliferative effect on human epidermoid carcinoma A431 cells.
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DOI：
10.1016/j.toxrep.2015.03.001
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发表时间：
2015
影响因子：
--
通讯作者：
Gudipalli P
中科院分区：
文献类型：
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作者：
Kamatham S;Kumar N;Gudipalli P
从银杏种皮中分离得到没食子酸（GA）和没食子酸甲酯（MG）。rottleriformis我们研究了纯化的GA和MG对A431皮肤癌细胞增殖的影响，并呈剂量和时间依赖性。我们分析了GA/MG处理的A431细胞中凋亡增强/抑制调节标记蛋白的剂量或时间依赖性。我们发现GA/MG是一种有效的天然分子，以避免皮肤癌的风险。没食子酸（GA）及其衍生物没食子酸甲酯（MG）是研究得很好的植物酚类化合物。它们在几种癌细胞系中表现出抗癌作用。然而，在Givotia rottleriformis种皮中存在GA/MG及其对人表皮样癌（A431）皮肤癌细胞的抑制作用未见报道。在这项研究中，我们已经分离和化学特性的生物活性化合物GA和MG从生物测定引导甲醇（MeOH）种皮提取物。rottleriformis将从开放式硅胶柱色谱法获得的级分进行体外酶测定。在7个组分中，只有组分5和组分6对考克斯-1和考克斯-2具有明显的抑制活性，其IC 50分别为28 μg/mL和9.3 μg/mL，IC 50分别为35 μg/mL和7.0 μg/mL。但是，我们没有发现5-LOX酶抑制活性。种皮中的主要成分为MG（10 mg/g DW）和GA（6 mg/g DW）。MTT法检测GA/MG对A431细胞生长的抑制作用，24 h和48 h的IC 50分别为25 μg/mL和53 μg/mL，11 μg/mL和43 μg/mL。Western blot分析表明，GA/MG处理后，随着GA/MG剂量的增加，Bcl-2表达下调，caspase-3表达上调。我们的研究结果得出结论，GA和MG具有潜在的抗癌作用，可用作皮肤癌的治疗剂。
We isolated gallic acid (GA) and methyl gallate (MG) from the seed coats of G. rottleriformis. We investigated the effect of purified GA and MG on A431 skin cancer cell proliferation in a dose and time dependent manner. We analyzed the apoptosis enhancer/suppressor regulatory marker proteins in GA/MG treated A431 cells in a dose or time dependent manner. We found that GA/MG is an effective natural molecule to avoid the risk of skin cancers. Gallic acid (GA) and its derivative methyl gallate (MG) are well studied plant phenolics. They have exhibited anticancer effects in several cancer cell lines. However, the presence of GA/MG in the seed coats of Givotia rottleriformis and their inhibitory effect on human epidermoid carcinoma (A431) skin cancer cells were not reported. In this study we have isolated and chemically characterized the bioactive compounds GA and MG from the bioassay guided methanolic (MeOH) seed coat extracts of G. rottleriformis. The fractions obtained from open silica column chromatography were subjected to in vitro enzymatic assays. Among seven fractions we found that only fractions 5 and 6 showed significant inhibition activity toward COX-1 with an IC50 value of 28 μg/mL and 9.3 μg/mL and COX-2 with an IC50 value of 35 μg/mL and 7.0 μg/mL respectively. However, we could not find 5-LOX enzyme inhibition activity. MG (10 mg/g DW) and GA (6 mg/g DW) were the major compounds of seed coats. Cell viability was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, which showed that GA/MG significantly reduced the growth of A431 cells with an IC50 value of 25 μg/mL and 53 μg/mL and 11 μg/mL and 43 μg/mL at 24 h and 48 h, respectively. However the cytotoxic effect of GA/MG on HaCaT normal skin keratinocyte cell line was found to be less. Western blot analysis has shown that GA/MG treatment down regulated Bcl-2 and up regulated cleaved caspase-3 with respect to increasing doses. Our results conclude that GA and MG have potential anticancer effects and can be used as therapeutic agents for skin cancers.