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目的:观察沙枣提取物对人肝癌Bel-7404细胞增殖抑制作用及其对细胞凋亡的影响。方法:以没食子酸为对照品,采用Folin-Ciocalteu法测定沙枣提取物各分离组分的总酚含量。通过MTT法,观察各组分对Bel-7404细胞增殖抑制作用。选取对Bel-7404增殖抑制活性最强的20%乙醇洗脱组分,设50、100、150μg/ml剂量组,采用Ainexin V-FITC/PI双染法,流式细胞仪检测其诱导细胞凋亡,并采用Hoechst 33258荧光染色法观察细胞形态变化。结果:沙枣提取物各分离组分对Bel-7404增殖抑制作用各不相同,其中20%乙醇洗脱组分的50、100、150μg/ml剂量组的凋亡率分别为29.8%、58.7%和76.2%;凋亡细胞的细胞核或细胞质内可见浓染致密的颗粒块状荧光。结论:沙枣提取物可抑制Bel-7404细胞增殖,诱导其凋亡,并与总酚含量呈现一定的量效关系。
OBJECTIVE: To observe the inhibitory effect of extracts of Elaeagnus angustifolia on the proliferation of human hepatocellular carcinoma Bel-7404 cells and its effect on apoptosis. Methods: Using gallic acid as reference substance, Folin-Ciocalteu method was used to determine the total phenolic content of each component of Radix extract. MTT assay was used to observe the inhibitory effect of each component on Bel-7404 cell proliferation. Selected 20% ethanol elution fractions with the strongest inhibitory activity on Bel-7404, 50, 100 and 150μg / ml dose groups were selected, and the apoptosis was induced by flow cytometry using Ainexin V-FITC / PI double staining The morphological changes of cells were observed by Hoechst 33258 fluorescence staining. Results: The inhibitory effects of different fractions of Radix extract on Bel-7404 proliferation were different. The apoptosis rates of 50, 100 and 150μg / ml 20% ethanol fractions were 29.8%, 58.7% And 76.2% respectively. Apoptotic cell nuclei or cytoplasm could be seen dense and dense granular lump fluorescence. Conclusion: Radix extract can inhibit the proliferation and induce the apoptosis of Bel-7404 cells, and show some dose-effect relationship with the total phenolics.