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鸡蛋黄蛋白质水解产物制备及其对MC3T3-E1细胞增殖分化的影响
引用本文:陈宏杰,金永国,马美湖.鸡蛋黄蛋白质水解产物制备及其对MC3T3-E1细胞增殖分化的影响[J].食品科学,2017,38(22):95-101.
作者姓名:陈宏杰  金永国  马美湖
作者单位:(华中农业大学食品科技学院,湖北?武汉 430070)
基金项目:公益性行业(农业)科研专项(201303084)
摘    要:鸡蛋黄经脱脂得到蛋黄蛋白质,直接以MC3T3-E1细胞增殖活力为指标,对水解条件进行优化,从而制备蛋黄蛋白质水解产物(egg yolk proteins hydrolysate,EYPH)并对其促MC3T3-E1细胞增殖分化的活性进行研究。结果表明:在对比胰蛋白酶与风味蛋白酶水解过程中,发现单一胰蛋白酶反应得到产物活性最好,胰蛋白酶反应的最佳水解条件为水解时间6 h、水解温度37℃、酶与底物质量比1∶50、p H 8.0。EYPH使MC3T3-E1细胞增殖活力得到提高,为129.89%,细胞周期分析得到S期细胞增加了4.69%,这2个方面证实EYPH对MC3T3-E1细胞的增殖活力有显著的提高。EYPH对MC3T3-E1细胞分化矿化有显著影响,碱性磷酸酶活力提高了9.1%;刺激骨钙素分泌,其含量上升2 mg/mL;形成更多的矿化结节。此外,EYPH还能刺激RUNX2特异性转录因子的基因表达,表达量提高了6.12倍。结果说明EYPH对成骨细胞MC3T3-E1增殖分化有显著的促进作用,因此,EYPH对于抗骨质疏松方面有较好的应用潜力,为开发其功能性食品提供一定的数据参考。

关 键 词:蛋黄蛋白质水解产物  水解条件  MC3T3-E1细胞  增殖  分化  

Preparation of Egg Yolk Protein Hydrolysate and Its Effect on Proliferation and Differentiation of MC3T3-E1 Cells
CHEN Hongjie,JIN Yongguo,MA Meihu.Preparation of Egg Yolk Protein Hydrolysate and Its Effect on Proliferation and Differentiation of MC3T3-E1 Cells[J].Food Science,2017,38(22):95-101.
Authors:CHEN Hongjie  JIN Yongguo  MA Meihu
Affiliation:(College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China)
Abstract:In this study, the hydrolysis conditions for preparing egg yolk protein hydrolysate (EYPH) from defatted egg yolk were optimized based on the viability of MC3T3-E1 cells when cultured in the presence of EYPH. Additionally, we also evaluated the promoting effect of EYPH on proliferation and differentiation of MC3T3-E1 cells. The results showed that the hydrolysis with trypsin alone yielded a product with stronger cell proliferation promoting activity than when it was used in combination with flavourzyme. The optimum hydrolysis conditions for trypsin were as follows: hydrolysis time, 6 h; temperature, 37 ℃; enzyme-to-substrate ratio, 1:50; and pH, 8.0. EYPH prepared under the optimized conditions increased the viability of MC3T3-E1 cells by 9.1% and the proportion of S-phase cells by 4.69% indicating that EYPH significantly increased the proliferation of MC3T3-E1 cells. EYPH significantly affected differentiation and mineralization of MC3T3-E1 cells as indicated by a 9.1% increase in alkaline phosphatase activity, an increase in osteocalcin secretion of 2 mg/mL, and the formation of more mineralized nodules of MC3T3-E1. Furthermore, EYPH also could stimulate the expression level of RUNX2 gene by 6.12 times. These finding showed that EYPH could significantly promote proliferation and differentiation in osteoblastic MC3T3-E1 cells. Therefore, EYPH has the potential to be used as a functional food ingredient for its anti-osteoporosis activity.
Keywords:egg yolk protein hydrolysate  hydrolysis conditions  MC3T3-E1 cells  proliferation  differentiation  
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