河蚬乙酸乙酯提取物的抗氧化活性及其对人胆管癌细胞的抑制作用

作为传统中药的河蚬具有退热、治疗肝病和解酒等功效.以新鲜河蚬 (Corbicula fluminea)肉为原料,经甲醇抽提,乙酸乙酯萃取等方法得到河蚬乙酸乙酯提取物.运用DPPH法检测其清除自由基的能力,通过MTT法和倒置显微镜观察其对人胆管癌细胞QBC939生长的影响,并通过流式细胞术分析其对QBC939细胞周期的影响.结果显示:河蚬乙酸乙酯提取物对DPPH自由基具有一定的清除作用,在较低的浓度下有较高的清除效果,IC50值为0.6 mg/mL,且随其浓度的增加而不断增强;河蚬乙酸乙酯提取物对QBC939细胞有显著的抑制作用,处理48 h,IC50值为250 μg/mL,细胞形态发生明显变化;处理后的QBC939细胞出现明显的凋亡峰.研究表明:河蚬乙酸乙酯提取物不但具有抗氧化活性,且对体外培养的QBC939细胞具有明显的增殖抑制作用,能够改变QBC939细胞周期分布,并诱导QBC939细胞发生凋亡.说明河蚬乙酸乙酯提取物可能具有天然药物开发的潜力,同时也为胆管癌的防治提供新的思路和方法.

Antioxidation Effect and Inhibition Effect on Human Cholangiocarcinoma Cells of Corbicula fluminea Ethyl Acetate Extract

As a traditional chinese herb, Corbicula fluminea has many effects, such as, defervescence, curing the liver ailment, antialcoholism, and so on. We used fresh Corbicula fluminea body as the material and obtained the ethyl acetate extract by the method of ethyl acetate extraction. The ability of antioxidation was measured by DPPH method. The inhibition rate of cholangiocarcinoma cells was measured by MTT method. The morphological change of QBC939 cells was detected by ordinary inverted microscopy. The effect on cell cycles of QBC939 was observed by flow cytometry (FCM) assay. The results showed that ethyl acetate extract from Corbicula fluminea has strong ability to scavenging up DPPH radical, of which the value of IC50 was 0.6 mg/mL. And it could also inhibit the growth of QBC939 cells in a dose- and time-dependent manner, and the IC50 value was determined to be 250 μg/mL for 48 h treatment. The morphological of QBC939 cells changed significantly. At the same time, FCM showed that Corbicula fluminea ethyl acetate extract could change the cell cycle of QBC939 cells, and induce apoptosis. These results indicated that Corbicula fluminea ethyl acetate extract had the ability of antioxidation and effectively inhibited the proliferation of the human cholangiocarcinoma cell line QBC939 in vitro and acted as an apoptosis inducer in its antitumor effects via inducing morphological changes and cell cycle arrest. Thus, Corbicula fluminea ethyl acetate extract has the potential of becoming a natural drug, and it also provide a new thought and a new method for the therapy of cholangiocarcinoma.