食品科学 ›› 2019, Vol. 40 ›› Issue (23): 60-65.doi: 10.7506/spkx1002-6630-20181017-179

• 基础研究 • 上一篇    下一篇

不同巴氏杀菌条件下的牛源乳铁蛋白对IEC-6细胞增殖的影响

席恩泽,赵晓,崔东影,盛雪,李梦寒,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:2019-12-15 发布日期:2019-12-24
  • 基金资助:
    “十二五”国家科技支撑计划项目(2013BAD18B00)

Effect of in Vitro Digestion Hydrolysate of Bovine Lactoferrin Subjected to Different Pasteurization Treatments on Proliferation of IEC-6 Cells

XI Enze, ZHAO Xiao, CUI Dongying, SHENG Xue, LI Menghan, XU Xiaoxi   

  1. (School of Food Science, Northeast Agricultural University, Harbin 150030, China)
  • Online:2019-12-15 Published:2019-12-24

摘要: 本研究以大鼠小肠隐窝上皮细胞(IEC-6)为对象,通过体外实验,采用3 种不同的巴氏杀菌方式(63 ℃、30 min,72 ℃、15 s和85 ℃、15 s)处理牛源乳铁蛋白,模拟婴儿胃肠道消化环境,利用所得的牛源乳铁蛋白水解液刺激IEC-6细胞,研究乳铁蛋白在巴氏杀菌后的免疫活性。结果表明:巴氏杀菌后的乳铁蛋白水解物仍具有促进肠细胞增殖的免疫学活性,且呈剂量依赖性。流式细胞实验结果证实巴氏杀菌后的牛源乳铁蛋白水解液可提高IEC-6细胞在G2/M期和S期的比例,并降低细胞凋亡率。反转录实时荧光定量聚合酶链式反应检测结果表明,牛源乳铁蛋白水解液提高cyclin B1、CDK1和cyclin E1的表达水平,降低p21的表达水平。综上可知,巴氏杀菌不能完全使乳铁蛋白免疫基团失活。两种巴氏杀菌方法(72 ℃、15 s和85 ℃、15 s),尤其是85 ℃、15 s处理,在保证食品安全的同时可最大限度地保留乳铁蛋白的免疫活性。3 种巴氏杀菌后的乳铁蛋白水解液促进肠道免疫的最佳质量浓度为0.1 mg/mL。

关键词: 乳铁蛋白, 巴氏杀菌, 肠道免疫

Abstract: After being subjected to different pasteurization treatments (63 ℃/30 min, 72 ℃/15 s and 85 ℃/15 s), bovine lactoferrin was digested in vitro in a simulated infant gastrointestinal environment. The resulting hydrolysates were separately used to challenge rat intestinal crypt epithelial cells (IEC-6) to evaluate their immunological activity. The results showed that all the three lactoferrin hydrolysates could promote small intestinal cell proliferation in a dose-dependent manner. Flow cytometry results demonstrated that these hydrolysates increased the percentage of IEC-6 cells in the G2/M and S phase and reduced cell apoptosis. The results of reverse transcription real-time quantitative PCR (RT-qPCR) indicated that the lactoferrin hydrolysates elevated the expression levels of cyclin B1, CDK1 and cyclin E1 and reduced the expression level of p21. Taken together, we found that that pasteurization could completely inactivate the active group of bovine lactoferrin. The two pasteurization treatments, 72 ℃/15 s and 85 ℃/15 s, especially the latter, could ensure food safety while maintaining the immunological activity of lactoferrin to the utmost extent. The best concentration to promote intestinal immunity was 0.1 mg/mL for all the lactoferrin hydrolysates.

Key words: lactoferrin, pasteurization, intestinal immunity

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