朝鲜族大酱中异黄酮的提取工艺优化及其抗肿瘤活性研究

为优化朝鲜族大酱异黄酮的提取工艺,并研究朝鲜族大酱中异黄酮的抗肿瘤活性,本实验用超声波法提取朝鲜族大酱中的异黄酮,通过单因素实验和正交实验优化其提取工艺,并通过CCK-8法对正常乳腺细胞、MDA-MB-231和MCF-7两种乳腺癌细胞的增殖抑制情况进行了测定,用Western Blot法检测了朝鲜族大酱中异黄酮处理后的两种乳腺癌细胞凋亡相关蛋白的表达情况。通过单因素和正交试验确定其最佳提取条件为提取时间80 min、料液比1:24、温度50 ℃,此时得率最高,为0.802%。CCK-8结果显示,不同浓度的朝鲜族大酱异黄酮对正常细胞无毒副作用,对MDA-MB-231和MCF-7两种乳腺癌细胞的增殖抑制情况均呈剂量-时间依赖性,且均有显著性差异(p<0.05)。对不同浓度朝鲜族大酱异黄酮处理过的两种细胞凋亡相关蛋白的检测结果显示,两种细胞随着异黄酮浓度的增加,其促凋亡蛋白Bax,caspase-3的表达量与空白对照组相比均显著上升(p<0.05),而抗凋亡蛋白Bcl-2的表达量显著下降(p<0.05),且呈现剂量依赖性。表明朝鲜族大酱异黄酮对乳腺癌细胞的生长能够起到抑制作用。

Study on Extraction Process Optimization and Antitumor Activity of Isoflavone from Korean Soybean Paste

In order to optimize the extraction process of isoflavones from Korean soybean paste and study the anti-tumor activity of isoflavones in Korean soybean paste,this experiment used ultrasonic method to extract isoflavones from Korean soybean paste,which was optimized by single factor experiment and orthogonal experiment. The extraction process was carried out,and the proliferation inhibition of normal breast cells,MDA-MB-231 and MCF-7 breast cancer cells was determined by CCK-8 method. Western Blot was used to detect the expression of apoptosis-related proteins in two breast cancer cells treated with isoflavone of Korean soybean paste. The optimum extraction conditions were determined by single factor and orthogonal experiments. The extraction time was 80 min,the ratio of material to liquid was 1∶24,and the temperature was 50 ℃. The yield was the highest at 0.802%. The results of CCK-8 showed that different concentrations of Korean soybean paste isoflavone had no toxic side effect on normal cells. The proliferation inhibition of MDA-MB-231 and MCF-7 breast cancer cells was dose-dependent and there were significant differences(p<0.05). The detection results of apoptosis-related proteins in two kinds of cells treated with different concentrations of Korean soybean paste isoflavone showed that the two kinds of cells increased with the concentration of isoflavone,the expression of Bax and caspase-3 significantly increased compared with the control group(p<0.05),while the expression of anti-apoptotic protein Bcl-2 significantly decreased(p<0.05). With dose dependence. It indicates that Korean soybean paste isoflavone can inhibit the growth of breast cancer cells.