摘要
本文研究乳酸菌发酵大麦过程中主要物质成分的变化及其发酵提取物（LFBE）对人结肠癌HT-29细胞增殖的抑制作用。利用植物乳杆菌（Lactobacillus plantarum dy-1,L.p）发酵大麦,测定发酵过程中蛋白质、多酚、总糖的含量变化;通过SDS-PAGE法观察发酵过程中蛋白质分子质量的变化情况。将LFBE作用于HT-29肿瘤细胞,采用CCK-8法和倒置荧光显微镜及扫描电镜观测细胞增殖能力与形态变化,通过Hoechst/PI双染法研究细胞凋亡,并用单细胞凝胶电泳检测细胞的DNA损伤。试验结果显示,0~48 h发酵过程中,LFBE的蛋白质含量先升后降,在36 h达到峰值15.41%,多酚含量由0.42%增至1.18%,总糖含量在12 h达到最高值56.82%,随后逐渐降低。大麦发酵后相对分子质量在32~37 ku范围的蛋白质含量显著增加。LFBE能够改变HT-29肿瘤细胞的形态,诱导其凋亡。结论：LFBE对人结肠癌HT-29细胞增殖具有显著的抑制作用,能够诱导HT-29细胞凋亡,导致其细胞解体。
To study the effect of fermented barley extract with lactic acid bacteria on inhibition effcets of human colon carcinoma cell strain HT-29, and research the changes of main components in fermentation process, Barly was fermented by Lactobacillus plantarum dy-1. Changes of protein, polyphenol and polysaccharide content in the fermentation process were analyzed by conventional methods, and protein molecular weight was obversed by SDS-PAGE. Cell growth inhibition was detected by CCK-8 assay, the cell morphology was observed by inverted microscope and scanning electron microscope, and the frequencies of apoptosis were determined by Hoechst/propidium iodide（PI） double staining method;Finally, we detected the DNA damage by single cell gel electrophoresis. The results showed that in the fermentation of48 h, the protein content of LFBE first increased and then decreased, and reached the maximum value of 15.41% after36 h of fermentation, polyphenol content increased from 0.42% to 1.18%, the polysaccharide content reached the maximum value of 56.82% after 12 h of fermentation and then decreased gradually. Electrophoresis results showed that the protein content that molecular weight was between 32 ~37 ku increased significantly after fermentation. Cell experiments indicated that LFBE could inhibited tumor cell proliferation, it could also change the morphology of tumor cells, induce tumor cell apoptosis, cause damage to DNA and even lead to the disintegration. We can draw a conclusion that LFBE can significantly inhibit human colon carcinoma cell strain HT-29 and induce cell apoptosis.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2016年第6期23-29,共7页
Journal of Chinese Institute Of Food Science and Technology
基金
江苏省产学研联合创新资金--前瞻性联合研究项目(BY2012172)
关键词
乳酸菌
大麦
发酵提取物
细胞凋亡
蛋白质
lactic acid bacteria
barley
fermentation extract
cell apoptosis
protein
