脂肪酸结合蛋白5对奶牛乳腺上皮细胞乳脂合成的影响

doi:10.16431/j.cnki.1671-7236.2017.04.009

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (4): 941-949.doi: 10.16431/j.cnki.1671-7236.2017.04.009

• 生物技术 • 下一篇

脂肪酸结合蛋白5对奶牛乳腺上皮细胞乳脂合成的影响

于萌萌, 李萌, 李姗姗, 李冬, 高学军

东北农业大学乳品科学教育部重点实验室, 哈尔滨 150030

收稿日期:2016-09-06 出版日期:2017-04-20 发布日期:2017-05-03
通讯作者: 高学军 E-mail:gaoxj5390@sina.com
作者简介:于萌萌(1992-),女,黑龙江双鸭山人,硕士生,研究方向:生物化学与分子生物学,E-mail:13115513593@163.com
基金资助:
国家高技术研究发展计划（863计划）项目（2013AA102504-03）

Effect of Fatty Acid Binding Protein 5 on Milk Fat Synthesis of Bovine Mammary Epithelial Cells

YU Meng-meng, LI Meng, LI Shan-shan, LI Dong, GAO Xue-jun

Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China

Received:2016-09-06 Online:2017-04-20 Published:2017-05-03

摘要/Abstract

摘要：

脂肪酸结合蛋白5（fatty acid binding protein 5，FABP5）是一种胞内脂质运载体。本试验应用甘油三酯测定法及BODIPY染色法检测奶牛乳腺上皮细胞（bovine mammary epithelial cell，BMEC）中甘油三酯及脂滴分泌情况，采用实时荧光定量PCR、Western blotting技术检测BMECs中FABP5对固醇调节元件结合蛋白-1c（SREBP-1c）的表达影响。结果显示，试验成功获得高纯度的BMECs，构建了pGCMV-IRES-EGFP-FABP5真核表达载体，且与空白对照组和空载体组相比，FABP5过表达组的甘油三酯和脂滴分泌量极显著增加（P<0.01），同时SREBP-1c及靶基因FAS、ACC的表达量均极显著增加（P<0.01）；试验成功筛选了FABP5 siRNA1为最佳干扰片段，当FABP5抑制时，抑制组的甘油三酯、脂滴分泌量极显著减少（P<0.01），SREBP-1c、FAS、ACC的表达量均极显著降低（P<0.01）。表明FABP5可通过上调SREBP-1c的表达从而促进BMEC中乳脂的合成。

关键词: 奶牛乳腺上皮细胞; 脂肪酸结合蛋白5; 固醇调节元件结合蛋白-1c; 乳脂

Abstract:

The fatty acid binding protein 5 (FABP5) is an intracellular lipid carrier. The TG GPO-POD assay kit and BODIPY staining methods were used to detect lipid secretion and triglyceride content,and the effect of FABP5 on SREBP-1c expression were detected by Real-time PCR and Western blotting methods. The results showed that high purity bovine mammary epithelial cells (BMECs) were successfully isolated and purified,and the eukaryotic expression vector pGCMV-IRES-EGFP-FABP5 was constructed in this experiment. Compared with the blank control group and empty vector group,the lipid secretion and triglyceride content, and the expression of SREBP-1c and FAS,ACC were extremely significantly increased when the FABP5 was overexpressed (P<0.01). FABP5 siRNA1 was selected as the optimal interference fragment, and when FABP5 was inhibited,the lipid secretion and triglyceride content,the expression of SREBP-1c,FAS and ACC were extremely significantly decreased (P<0.01).The results indicated that FABP5 could promote the synthesis of milk fat in BMEC by up-regulating the expression of SREBP-1c.

Key words: bovine mammary epithelial cells (BMEC); FABP5; SREBP-1c; milk fat

中图分类号:

S823.9⁺1

于萌萌, 李萌, 李姗姗, 李冬, 高学军. 脂肪酸结合蛋白5对奶牛乳腺上皮细胞乳脂合成的影响[J]. 《中国畜牧兽医》, 2017, 44(4): 941-949.

YU Meng-meng, LI Meng, LI Shan-shan, LI Dong, GAO Xue-jun. Effect of Fatty Acid Binding Protein 5 on Milk Fat Synthesis of Bovine Mammary Epithelial Cells[J]. , 2017, 44(4): 941-949.

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脂肪酸结合蛋白5对奶牛乳腺上皮细胞乳脂合成的影响

Effect of Fatty Acid Binding Protein 5 on Milk Fat Synthesis of Bovine Mammary Epithelial Cells

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