| FADS2基因在奶牛乳腺细胞中的过表达和干扰研究 |
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| 引用本文： | 马小娅,庞春英,邓廷贤,段安琴,陆杏蓉,梁莎莎,梁贤威. FADS2基因在奶牛乳腺细胞中的过表达和干扰研究[J]. 中国畜牧兽医, 2019, 46(3): 652-660. DOI: 10.16431/j.cnki.1671-7236.2019.03.002 |
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| 作者姓名： | 马小娅 庞春英 邓廷贤 段安琴 陆杏蓉 梁莎莎 梁贤威 |
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| 作者单位： | 中国农业科学院广西水牛研究所, 农业部(广西)水牛遗传繁育重点实验室, 南宁 530001 |
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| 基金项目： | 广西水产畜牧科技推广应用项目（桂渔牧科201633009）；广西水牛研究所基本科研业务费项目（1705002）；国家国际科技合作专项项目（2014DFA31970）；广西国际合作项目（贵科合15104001-3）；广西重大科技专项（桂科AA16450002） |
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| 摘 要： | 为了研究脂肪酸脱氢酶2(fatty acid desaturases 2,FADS2)基因在奶牛乳腺细胞脂肪酸代谢中的作用,本研究在奶牛乳腺上皮细胞中对FADS2基因进行过表达和干扰,研究FADS2基因表达对脂肪酸合成相关基因的调控及对奶牛乳腺上皮细胞中甘油三酯含量的影响。针对FADS2基因的CDS序列设计siRNA和过表达载体pcDNA3.1-FADS2-EGFP,转染奶牛乳腺细胞检测FADS2基因过表达和干扰对脂肪酸代谢相关基因表达的影响及细胞中甘油三酯含量的变化。结果显示,试验成功获得过表达载体pcDNA3.1-FADS2-EGFP和干扰片段,转染细胞后具有良好的过表达和干扰效果。FADS2基因过表达后,1-酰基甘油磷酸酰基转移酶(AGPAT1)、固醇调节元件结合蛋白裂解激活蛋白(SCAP)、3-磷酸甘油转移酶(GPAM)、脂肪酸延长链5(ELOVL5)、乙酰辅酶A酰基转移酶1(ACAA1)、脂肪酸脱氢酶1(FADS1)、二酰基甘油转酰基酶1(DGAT1)和过氧化物酶体增殖激活受体α(PPARα)基因显著下调(P<0.05),脂滴蛋白2(PLIN2)基因极显著上调(P<0.01)。FADS2基因干扰过后可引起AGPAT1、GPAM、ELOVL5、ACAA1、PLIN2和FADS1基因显著上调(P<0.05),脂肪酸合成胰岛素诱导基因1(INSIG1)极显著上调(P<0.01),DGAT1和PPARα基因显著下调(P<0.05)。甘油三酯检测结果显示,FADS2基因过表达和干扰均可降低奶牛乳腺上皮细胞中甘油三酯的含量。综上所述,在奶牛乳腺上皮细胞中,FADS2基因能调控脂质合成相关基因的表达,对乳腺脂质合成具有调控作用。
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| 关 键 词： | 奶牛乳腺细胞 FADS2基因 过表达 干扰 脂肪酸合成 |
| 收稿时间： | 2018-08-27 |
Overexpression and Interference of FADS2 Gene in Mammary Gland Cells of Dairy Cows |
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| MA Xiaoya,PANG Chunying,DENG Tingxian,DUAN Anqin,LU Xingrong,LIANG Shasha,LIANG Xianwei. Overexpression and Interference of FADS2 Gene in Mammary Gland Cells of Dairy Cows[J]. China Animal Husbandry & Veterinary Medicine, 2019, 46(3): 652-660. DOI: 10.16431/j.cnki.1671-7236.2019.03.002 |
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| Authors: | MA Xiaoya PANG Chunying DENG Tingxian DUAN Anqin LU Xingrong LIANG Shasha LIANG Xianwei |
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| Affiliation: | Key Laboratory of Buffalo Genetics, Breeding and Reproduction Technology, Minstry of Agriculture(Guangxi), Guangxi Buffalo Research Institute, Chinese Academy Academy of Agricultural Science, Nanning 530001, China |
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| Abstract: | In order to explore the role of fatty acid desaturases 2 (FADS2) gene in fatty acid metabolism in dairy mammary gland cells,this study overexpressed and interfered with FADS2 gene in dairy mammary epithelial cells,and studied the regulation of FADS2 gene on fatty acid synthesis related genes and the effect on triglyceride content in dairy mammary epithelial cells.The siRNA and overexpression vector pcDNA3.1-FADS2-EGFP were designed according to the CDS sequence of FADS2 gene.The effect of FADS2 gene overexpression and interference on the expression of fatty acid metabolism related genes and the change of triglyceride content in cells were detected by transfecting mammary gland cells.The results showed that the overexpression vector pcDNA3.1-FADS2-EGFP and the interference fragment were successfully obtained,and the transfected cells had good overexpression and interference effects.After overexpression of FADS2 gene,AGPAT1,SCAP,GPAM,ELOVL5,ACAA1,FADS1,DGAT1 and PPARα genes were significantly down-regulated (P<0.05),and PLIN2 gene was extremely significantly up-regulated (P<0.01).After FADS2 gene interference,AGPAT1,GPAM,ELOVL5,ACAA1,PLIN2 and FADS1 genes were significantly up-regulated (P<0.05),INSIG1 gene was extremely significantly up-regulated (P<0.01),andDGAT1 and PPARα genes were significantly down-regulated (P<0.05).Triglyceride assay showed that FADS2 gene overexpression and interference could reduce the content of triglyceride in mammary gland epithelial cells.In summary,FADS2 gene could regulate the expression of lipid synthesis-related genes in mammary gland epithelial cells,and had a regulatory effect on breast lipid synthesis. |
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| Keywords: | mammary epithelial cells FADS2 gene overexpression interfere fatty acid synthesis |
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