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林蛙油蛋白酶水解物的免疫促进功能与抗氧化活性

ISSN号：1005-9989
期刊名称：食品科技
时间：2014.3.20
页码：211-214
分类：R285[医药卫生—中药学;医药卫生—中医学]
作者机构：[1]北京林业大学生物科学与技术学院,北京100083, [2]北京林业大学林业食品加工与安全北京市重点实验室,北京100083
相关基金：基金项目：国家自然科学基金资助子项目（J1103516-2D26）;林业公益性行业科研专题（201004081）.
相关项目：北京林业大学生物学基地

关键词：山杏仁蛋白, 人结肠癌细胞, 抑制细胞增殖, 细胞死亡, 乙醇, apricot kernel protein, human colon carcinoma cells, inhibited cell proliferation, cell death, ethanol

中文摘要：

分别以水、盐、乙醇及NaOH溶液为提取剂，从山杏仁中提取蛋白，通过细胞增殖分析实验，研究山杏仁蛋白对人结肠癌细胞Caco-2生长的影响。结果表明，用有机溶剂乙醇提取的山杏仁蛋白显著抑制Caco-2细胞生长（P〈0．01，其IC50值为1．018mg／mL），而用其他提取剂提取的山杏仁蛋白并没有显示同样活性。以乙二醇、异丙醇及叔丁醇等其他有机溶剂代替乙醇提取的山杏仁蛋白，也没有显示抑制Caco-2细胞生长作用。因用乙醇提取的山杏仁蛋白具有显著抑制Caco-2细胞生长作用，称为活性山杏仁蛋白。SDS-聚丙烯凝胶电泳分析结果表明，21ku和41ku2个亚基存在于活性山杏仁蛋白中。细胞活力检测结果表明，用5mg／mL浓度活性山杏仁蛋白处理Caco-2细胞48h后，死细胞占细胞总数的89．5％。TUNEL和AnnexinV—FITC染色等细胞凋亡检测结果分别表明，活性山杏仁蛋白诱导了Caco-2细胞凋亡。

英文摘要：

Proteins were extracted from apricot kernel by water, salt, ethanol and NaOH solution respectively. The effect of the proteins on growth of Caco-2 human colon adenocarcinoma cell was studied by cell proliferation assay. The results indicated that the protein extracted from apricot kernel by ethanol inhibitedthe proliferation of the cells （P〈0.01, IC50 value is 1.018 mg/mL） significantly, while the ones from apricot kernel by other solution did not show the same activity. Replacing ethanol by ethylene glycol, isopropanol and tert-butanol, the extracts were almost inert. The protein extracted by ethanol was called active apricot kernel protein, due to its inhibiting effect on Caco-2 cell proliferation. SDS-polyacrylamide gel electrophoresis assay showed that 21 ku and 42 ku two protein subunits existing in the active apricot kernel protein. Cell viability assay using trypan blue solution indicated that 89.5% of Caco-2 cells died after exposing 5 mg/mL active apricot kernel protein for 48 h. TUNEL and Annexin V-FITC staining assays both showed that active apricot kernel protein induced Caco-2 cells apoptosis.

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