<em>SLC27A6</em>抑制表达对奶牛乳腺上皮细胞脂质代谢的影响

doi:10.3969/j.issn.1674-7968.2021.09.008

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摘要牛奶中的长链脂肪酸(long chain fatty acids, LCFA)尤其是长链多不饱和脂肪酸(polyunsaturated fatty acids, PUFA)有助于降低人类(Homo sapiens)患心血管疾病、癌症、二型糖尿病的风险。但目前对乳腺组织中LCFA合成、代谢的分子机制知之甚少。脂肪酸转运蛋白6 (solute carrier family 27 member 6, SLC27A6)的主要功能是促进细胞从外界摄取LCFA,进而参与细胞内的各类脂质代谢。但是目前SLC27A6与乳腺脂质代谢基因之间的调控关系尚不明确。本研究利用siRNA干扰技术干预奶牛(Bos taurus)乳腺上皮细胞(bovine mammary epithelial cells, BMECs)中SLC27A6的表达,采用荧光定量PCR分析乳腺细胞中SLC27A6和脂质代谢相关基因的表达,利用Western blot技术分析了蛋白的表达变化,并对细胞内甘油三酯含量和脂肪酸组成的变化进行分析。结果显示,与对照组相比,转染siRNA SLC27A6-Bos-915后,SLC27A6基因和蛋白质的表达量明显下降,细胞内参与脂肪酸激活和转运的关键基因长链脂肪酸乙酰辅酶A合成酶4基因(long-chain acyl-CoA synthetase 4, ACSL4)和脂肪酸转运蛋白基因CD36、脂肪氧化相关基因肉毒碱棕榈酰转移酶基因(carnitine palmitoyl transferase 1A, CPT1A)的表达量下降,过氧化物酶体增生激活受体γ (peroxisome proliferative activated receptor gamma, PPARG)、脂肪酸结合蛋白(fatty acid binding protein 3, FABP3)基因及脂肪酸去饱和酶(fatty acid desaturase 2, FADS2)基因的表达量上升。与此同时,细胞内的甘油三酯含量上升,棕榈酸(C16∶0)和硬脂酸(C18∶0)的含量降低,油酸(C18∶1cis9)和花生四烯酸(C20∶4cis5,8,11,14)的含量上升。本研究可为解析SLC27A6在调节奶牛乳腺脂质代谢中的机制提供参考,为实现乳成分的定向遗传调控,生产优质原料奶提供理论依据。

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	沈子亮
	姜惠
	陈代杰
	韩紫音
	毛永江
	李明勋
	杨章平
	张慧敏

关键词 ：脂肪酸转运蛋白6 (SLC27A6), 奶牛, 乳腺上皮细胞, 脂质代谢

Abstract：Fatty acid compositions are crucial indicators of milk quality in dairy cow. Long chain fatty acid (LCFA), especially polyunsaturated fatty acids (PUFA) are beneficial in reducing the risk of cardiovascular disease, cancer, and type 2 diabetes in humans (Homo sapiens). However, the metabolic mechanism of LCFA in mammary gland is still unknown. Solute carrier family 27 member 6 (SLC27A6) is a transmembrane protein that could enhance the uptake of LCFA into cells, thus regulating downstream fatty acid (FA) metabolic pathways. But the relationship between SLC27A6 and fatty acid metabolic genes is still unclear. In this study, SLC27A6 knockdown in bovine (Bos taurus) mammary epithelial cells (BMECs) was accomplished by RNA interference to further assess the function of SLC27A6. The relative mRNA expression level of genes related to lipid metabolism after SLC27A6 knockdown in BMECs was analyzed by real-time quantitative PCR, and the protein level of SLC27A6 was analyzed by Western blot. The effect of SLC27A6 knockdown on triglyceride (TG) content and FA composition in BMECs were determined.The result showed that the mRNA and protein levels of SLC27A6 were decreased in SLC27A6-Bos-915 group compared with the control group. And knockdown of SLC27A6 dramatically downregulated the mRNA abundance of genes associated with FA activation long-chain acyl-CoA synthetase 4 (ACSL4), fatty acid transporter CD36 and oxidation carnitine palmitoyl transferase 1A (CPT1A), while abundance of genes associated with transcription regulation peroxisome proliferative activated receptor gamma (PPARG), fatty acid binding protein 3 (FABP3), and fatty acid desaturase 2 (FADS2) were upregulated. In addition, SLC27A6 was silenced, the intracellular content of TG and the percentage of oleic acid (C18∶1cis9) and arachidonic acid (C20∶4cis5,8,11,14) were higher, whereas that of palmitic acid (C16∶0) and stearic acid (C18∶0) were lower. Overall, these results could provide strong support for a central role of SLC27A6 in the regulation of lipid metabolism in BMECs, and have an important guiding significance to improving milk quality by breeding programs.

Key words： Solute carrier family 27 member 6 (SLC27A6) Dairy cow Bovine mammary epithelial cells Lipid metabolism

收稿日期: 2020-12-30

ZTFLH:

S823

基金资助:国家自然科学基金青年基金(31702142); 扬州大学大学生科技创新基金(X20210671)

通讯作者: * minmin-911@163.com

引用本文:

沈子亮, 姜惠, 陈代杰, 韩紫音, 毛永江, 李明勋, 杨章平, 张慧敏. SLC27A6抑制表达对奶牛乳腺上皮细胞脂质代谢的影响[J]. 农业生物技术学报, 2021, 29(9): 1733-1740.
SHEN Zi-Liang, JIANG Hui, CHEN Dai-Jie, HAN Zi-Yin, MAO Yong-Jiang, LI Ming-Xun, YANG Zhang-Ping, ZHANG Hui-Min. Effect of SLC27A6 Expression Inhibition on Lipid Metabolism in Bovine (Bos taurus) Mammary Epithelial Cells. 农业生物技术学报, 2021, 29(9): 1733-1740.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.09.008 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I9/1733

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