脂肪酸结合蛋白5对奶牛乳腺上皮细胞乳脂合成的影响

发布时间：2018-02-22 15:01

本文关键词： 奶牛乳腺上皮细胞脂肪酸结合蛋白固醇调节元件结合蛋白-c 乳脂　出处：《中国畜牧兽医》2017年04期 　论文类型：期刊论文

【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(P0.01),同时SREBP-1c及靶基因FAS、ACC的表达量均极显著增加(P0.01);试验成功筛选了FABP5siRNA1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(P0.01),SREBP-1c、FAS、ACC的表达量均极显著降低(P0.01)。表明FABP5可通过上调SREBP-1c的表达从而促进BMEC中乳脂的合成。
[Abstract]:Fatty acid binding protein acid binding protein 5 (FABP5) is an intracellular liposome. Triglyceride and lipid droplet secretion in bovine mammary epithelial cells were detected by triglyceride assay and BODIPY staining. The effect of FABP5 on the expression of steroid regulatory element binding protein SREBP-1c in BMECs was detected by real-time fluorescence quantitative blotting. The results showed that high purity BMECs were obtained successfully and the eukaryotic expression vector of pGCMV-IRES-EGFP-FABP5 was constructed. Compared with the blank control group and empty vector group, the secretion of triglyceride and lipid droplets in the overexpression group of FABP5 was significantly increased, and the expression of SREBP-1c and FAS-ACC were significantly increased. The FABP5siRNA1 was selected as the best interfering fragment when FABP5 was inhibited. The secretion of triglyceride and lipid droplets significantly decreased the expression of SREBP-1FASA in BMEC. It was suggested that FABP5 could promote the synthesis of milk fat in BMEC by upregulating the expression of SREBP-1c.
【作者单位】：东北农业大学乳品科学教育部重点实验室;
【基金】：国家高技术研究发展计划(863计划)项目(2013AA102504-03)
【分类号】：S823.91

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