食品与发酵工业 >

2021 , Vol. 47 >Issue 11: 61 - 67

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026326

研究报告

鼠李糖乳杆菌hsryfm 1301发酵乳在非酒精性脂肪肝细胞模型中对人肝细胞L-02的益生作用

  • 陈大卫 ,
  • 梁娇娇 ,
  • 程月 ,
  • 瞿恒贤 ,
  • 陈春萌 ,
  • 任晨瑜 ,
  • 张臣臣 ,
  • 关成冉 ,
  • 马文龙 ,
  • 陈霞 ,
  • 李启明 ,
  • 顾瑞霞
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  • 1(扬州大学 食品科学与工程学院 江苏省乳品生物技术与安全控制重点实验室,江苏 扬州,225127)
    2(新希望乳业股份有限公司,四川 成都,610023)
博士,副教授(顾瑞霞教授为通讯作者,E-mail: guruixia1963@163.com)

收稿日期: 2020-12-01

  修回日期: 2021-02-04

  网络出版日期: 2021-07-16

基金资助

国家自然科学基金青年项目(31701627,31801565);国家自然科学基金面上项目(31972094);江苏省高等学校自然科学研究重大项目(19KJA140004);江苏省高等学校自然科学研究面上项目(17KJB550009);“十三五”国家重点研发计划课题项目(2019YFF0217602);成都市重大科技应用示范项目(2019-YF09-00055-SN)

收起

Probiotic function of Lactobacillus rhamnosus hsryfm 1301 fermented milk on human hepatocyte L-02 in NAFLD cell model

  • CHEN Dawei ,
  • LIANG Jiaojiao ,
  • CHENG Yue ,
  • QU Hengxian ,
  • CHEN Chunmeng ,
  • REN Chenyu ,
  • ZHANG Chenchen ,
  • GUAN Chengran ,
  • MA Wenlong ,
  • CHEN Xia ,
  • LI Qiming ,
  • GU Ruixia
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  • 1(Jiangsu Province Key Lab of Dairy Biotechnology and Safety Control,College of Food Science and Engineering,Yangzhou University; Yangzhou 225127,China)
    2(New Hope Dairy Co.Ltd.,Chengdu 610023,China)

Received date: 2020-12-01

  Revised date: 2021-02-04

  Online published: 2021-07-16

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摘要

建立由游离脂肪酸(free fat acid,FFA)诱导的体外人肝细胞L-02 非酒精性脂肪性肝病(nonalcoholic fatty liver disease,NAFLD)模型,探究益生菌发酵乳在模型中对人肝细胞的益生作用。利用FFA(油酸:棕榈酸=2:1,摩尔比)诱导人肝细胞脂肪变性,通过细胞存活率、胞内甘油三酯(triglyceride,TG)和总胆固醇(total cholesterol,TC)含量、培养液中天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)和谷丙转氨酶(alanine aminotransferase,ALT)含量及细胞凋亡率等指标来评价脂肪变性模型,探讨鼠李糖乳杆菌hsryfm 1301发酵乳干预大鼠4周后的血清在模型中对人肝细胞L-02的影响。结果表明,当模型中FFA的浓度为1.5和2 mmol/L时,人肝细胞的存活率显著低于未添加(P＜0.05),均低于32%,细胞数量明显减少且形状发生变化;当FFA浓度为1 mmol/L时,人肝细胞存活率较高,为86.37%,脂滴数量达到最高,此时建立的NAFLD细胞模型中胞内TG含量和培养液中AST含量分别为5.73 mmol/L和113.50 U/L,均显著高于未添加(P＜0.05),而胞内TC含量及培养液中ALT含量则无显著性差异(P＞0.05),且细胞凋亡率差异较小。鼠李糖乳杆菌hsryfm 1301发酵乳干预大鼠后的血清降低了NAFLD模型中肝细胞的脂滴数量,胞内TG含量及大鼠血清细胞培养液中AST含量均显著低于未干预菌株的对照组大鼠血清(P＜0.05),分别为1.95 mmol/L和93.33 U/L;细胞凋亡率较对照组下降了2.96%,正常细胞数量上升了4.50%。利用1 mmol/L的FFA可以建立人肝细胞L-02的NAFLD模型,鼠李糖乳杆菌hsryfm 1301发酵乳干预后的大鼠血清对模型中人肝细胞L-02的脂肪变性具有改善作用。

关键词： 鼠李糖乳杆菌hsryfm 1301; 发酵乳; 人肝细胞L-02; 游离脂肪酸; 非酒精性脂肪性肝病

本文引用格式

陈大卫 , 梁娇娇 , 程月 , 瞿恒贤 , 陈春萌 , 任晨瑜 , 张臣臣 , 关成冉 , 马文龙 , 陈霞 , 李启明 , 顾瑞霞 . 鼠李糖乳杆菌hsryfm 1301发酵乳在非酒精性脂肪肝细胞模型中对人肝细胞L-02的益生作用[J]. 食品与发酵工业, 2021 , 47(11) : 61 -67 . DOI: 10.13995/j.cnki.11-1802/ts.026326

Abstract

A nonalcoholic fatty liver disease (NAFLD) model of human hepatocytes L-02 in vitro was established by free fat acid (FFA) , and the beneficial effect of fermented milk of Lactobacillus rhamnosus hsryfm 1301 on L-02 cells in the model was explored. Free fatty acids (mol ratio, oleic acid∶palmitic acid=2∶1) were used to induce human hepatocytes steatosis, and the model was evaluated by the survival rate of cells, the content of total triglyceride (TG), total cholesterol (TC) and lipid droplets in cells, the content of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the culture medium of L-02 cells, and the apoptosis rate of cells. The effect of rats' serum, which was obtained after intervention by fermented milk of L. rhamnosus hsryfm 1301 for 4 weeks, on the L-02 cells in the model was discussed. The results showed that the survival rate of L-02 cells were all lower than 32% when the concentration of FFA was 1.5 mmol/L and 2 mmol/L in the model, which were significantly lower than that without FFA (P<0.05), and the number of L-02 cells was reduced obviously and the shape changed. While the survival rate of L-02 cells was 86.37% when the concentration was 1 mmol/L, and the number of lipid droplets was the highest, and the content of TG in the L-02 cells and the content of AST in the cell culture medium were significantly higher than that of the control group (P<0.05), which were 5.73 mmol/L and 113.50 U/L respectively. The apoptosis rate of L-02 cells, the content of TC in the L-02 cells and the content of ALT in the cell culture medium had no significant difference (P>0.05). The number of lipid droplets in the L-02 cells was reduced, and the content of TG in the L-02 cells and the content of AST in the cell culture medium were significantly decreased by the serum of rats that were administered by the fermented milk of L. rhamnosus hsryfm 1301 when compared with the serum of rats that were not administered (P<0.05), which were 1.95 mmol/L and 93.33 U/L respectively. And the apoptosis rate was decreased by 2.96%, the number of normal cells was increased by 4.5%. The NAFLD model of human hepatocyte L-02 in vitro can be established by 1 mmol/L FFA. The human hepatocyte L-02 in NAFLD model can be improved by the serum of rats that were intervened by the fermented milk of L. rhamnosus hsryfm 1301.

Key words： Lactobacillus rhamnosus hsryfm 1301; fermented milk; human hepatocyte L-02; free fatty acids; nonalcoholic fatty liver disease

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