食品科学 ›› 2020, Vol. 41 ›› Issue (1): 139-146.doi: 10.7506/spkx1002-6630-20181119-211

• 营养卫生 • 上一篇    下一篇

不同热处理方式的α-乳白蛋白对正常人肠道上皮细胞株增殖、周期及凋亡的影响

盛雪,张居典,李梦寒,崔东影,席恩泽,许晓曦   

  1. (东北农业大学食品学院,黑龙江 哈尔滨 150030)
  • 出版日期:2020-01-15 发布日期:2020-01-19
  • 基金资助:
    “十二五”国家科技支撑计划项目(2013BAD18B00)

Effect of α-Lactalbumin with Different Heat Treatments on Proliferation, Cell Cycle and Apoptosis of Human Intestinal Epithelial Cells

SHENG Xue, ZHANG Judian, LI Menghan, CUI Dongying, XI Enze, XU Xiaoxi   

  1. (School of Food Science, Northeast Agriculture University, Harbin 150030, China)
  • Online:2020-01-15 Published:2020-01-19

摘要: 以经过巴氏杀菌方式(63?℃加热30?min、72?℃加热15?s、85?℃加热15?s、95?℃加热10?min)处理的α-乳白蛋白(α-lactalbumin,α-LA)为对象,研究其对正常人肠道上皮细胞株(human?intestinal?epithelial?cells,HIEC)增殖、周期及凋亡的影响,并比较不同热处理对HIEC作用效果的差异性。在通过圆二色光谱测定α-LA二级结构、分析不同热处理方式对α-LA二级结构影响的基础上,模拟婴儿肠道条件对α-LA进行体外消化,采用CCK-8法和流式细胞术检测热处理消化后α-LA作用HIEC?48?h后,对细胞增殖、细胞周期以及细胞凋亡的影响。结果显示:经不同加热处理及未经热处理的消化后α-LA均对HIEC增殖具有促进作用,且在一定范围内促进效果随α-LA质量浓度升高而加强,在α-LA质量浓度为0.10?mg/mL、72?℃加热15?s时促进效果最为显著(P<0.05);经不同加热处理及未经热处理的消化后α-LA在质量浓度为0.10?mg/mL、作用HIEC 48?h后,G0/G1期细胞比例显著降低(P<0.05),S期和G2/M期细胞比例显著升高(P<0.05),细胞凋亡率显著降低(P<0.05)。综上,热处理α-LA可以明显促进HIEC增殖,其作用机制可能与分裂细胞的比例增加从而抑制细胞凋亡有关。α-LA应用于婴儿配方产品中或可提高婴儿免疫能力,推荐添加量为0.10?mg/mL,推荐热处理方式为72?℃加热15?s,但仍需进一步研究。

关键词: α-乳白蛋白, 正常人肠道上皮细胞株, 二级结构, 肠道免疫

Abstract: The effects of α-lactalbumin (α-LA) treated by pasteurization (63 ℃/30 min, 72 ℃/15 s, 85 ℃/15 s, or 95℃/10 min) on the proliferation, cell cycle and apoptosis of normal human intestinal epithelial cells (HIEC) were studied and compared. The secondary structure of α-LA with different pasteurization treatments was determined by circular dichroism spectroscopy. Then, the pasteurized α-LA was digested in vitro under simulated infant intestinal conditions and the effect of the resulting α-LA hydrolysates on the proliferation, cell cycle and apoptosis of HIEC cells at 48 h of culture was detected by cell counting kit-8 (CCK-8) assay and flow cytometry. The results showed that both raw and pasteurized α-LAs could significantly promote the proliferation of HIEC cells in a concentration-dependent manner in a certain range, and the most significant (P < 0.05) effect was achieved under the conditions of pasteurization at 72 ℃ for 15 s and 0.10 mg/mL concentration. Flow cytometry showed that both digested pasteurized α-LA and digested α-LA at a concentration of 0.10 mg/mL significantly reduced the percentage of HIEC cells in the G0/G1 phase (P < 0.05), increased the percentage of HIEC cells in the G2/M and S phase (P < 0.05), and decreased cell apoptosis (P < 0.05). These results indicated that pasteurized α-LA could significantly promote the proliferation of HIEC cells, and its mechanism may be related to the increase of the proportion of dividing cells and the consequent inhibition of apoptosis. The supplementation of α-LA pasteurized at 72 ℃ for 15 s at a recommended concentration of 0.10 mg/mL to infant formulas may improve immune function in infants, which needs to be further explored.

Key words: α-lactalbumin, normal human intestinal epithelial cells, secondary structure, intestinal immunity

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