N-butanol fraction of Entada phaseoloides ethanol extract inhibits hepatocellular carcinoma HepG2 cell proliferation by inducing apoptosis.
N-butanol fraction of Entada phaseoloides ethanol extract inhibits hepatocellular carcinoma HepG2 cell proliferation by inducing apoptosis.
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发表时间：
2014-04
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通讯作者：
Li Zhang;Ling Huang;Qiang-Ni Liu;Shao-Yi Kuang;Qiao-Qin Xu;Xianjin Qin;S. Fu;Qian Wu;Qibing Liu;Yu-qin Zeng
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作者：
Li Zhang;Ling Huang;Qiang-Ni Liu;Shao-Yi Kuang;Qiao-Qin Xu;Xianjin Qin;S. Fu;Qian Wu;Qibing Liu;Yu-qin Zeng
目的从药用植物山龙眼（Heliciopsis lobata（Merr.）本研究对五种植物的药理作用机制进行了初步探讨，并对五种植物的药理作用机制进行了初步研究。方法采用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四氮唑（MTT）比色法检测植物提取物对HepG 2细胞增殖的抑制作用。Hoechst 33342染色检测细胞凋亡，JC-1探针检测线粒体跨膜电位（ΔCgr;m）。结果与莲子草相比，裂叶山慈姑、鬼针草、车前草和菝葜提取物对HepG 2细胞增殖的抑制作用较弱（均p<0.05）。结果表明，四棱豆藤乙醇提取物正丁醇部位的抑菌率最高。用500、250和100 μg/ml正丁醇提取物处理HepG 2细胞，分别导致细胞活力下降89.92± 0.58%，IC 50为81.66± 0.42%，68.85±0.57%，表明IC 50为9.27 μg/ml。在100 μg/ml的恩他达正丁醇提取物存在下24 h，在HepG 2细胞中观察到凋亡的细胞核和深染的、致密的荧光块状颗粒，这种效应随着提取物浓度的增加而增加。最后，我们表明，恩塔达phaseoloides正丁醇提取物诱导线粒体膜电位的去极化。结论莲子草正丁醇提取物可能通过线粒体凋亡途径诱导细胞凋亡，从而抑制HepG 2细胞增殖。因此，该提取物是发现用于治疗HCC的新候选药物的潜在天然来源。
PURPOSE To screen for substances with inhibitory effects on the proliferation of hepatocellular carcinoma (HCC) HepG2 cell line from extracts of traditional Chinese medicinal plants including Heliciopsis lobata (Merr.) Sleum, Bidens pilosa, Entada phaseoloides, Plantago major, and Smilax, and unveil their mechanism of action. METHODS 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was used to assess the inhibition of HepG2 cell proliferation by plant extracts. Cell apoptosis was evaluated by Hoechst 33342 staining and mitochondrial transmembrane potential (ΔCgr;m) dissipation was measured using JC-1 probe by fluorescence microscopy. RESULTS Heliciopsis lobata, Bidens, Plantago, and Smilax extracts showed reduced inhibitory effects on HepG2 cell proliferation compared with Entada phaseoloides (all p<0.05). The n-butanol fraction of Entada phaseoloides ethanol extract exhibited the highest inhibition rate. Treatment of HepG2 cells with 500, 250, and 100 μg/ml n-butanol extract resulted in 89.92±0.58%, IC50 81.66±0.42%, 68.85±0.57% decrease in cell viability, respectively, indicating an IC50 of 9.27 μg/ml. In the presence of 100 μg/ml entada phaseoloides n-butanol extract for 24h, apoptotic nuclei and hyperchromatic, dense fluorescent massive granules were observed in the cytoplasm, effects that increased with extract concentrations in HepG2 cells. Finally, we showed that Entada phaseoloides n-butanol extract induced depolarization of mitochondrial membrane potential. CONCLUSIONS Entada phaseoloides n-butanol extract inhibits HepG2 cell proliferation by inducing cell apoptosis likely through mitochondrial apoptotic pathway. This extract is therefore a potential natural source towards the discovery for a new drug-candidate for the treatment of HCC.