Fan Xinyang, Zhu Wei, Qiu Lihua, Zhang Guangle, Zhang Yongyun, Miao Yongwang
Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming, China.
College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China.
J Anim Physiol Anim Nutr (Berl). 2022 Jan;106(1):1-11. doi: 10.1111/jpn.13536. Epub 2021 Mar 19.
Recent studies have shown elongase of very-long-chain fatty acids 6 (ELOVL6) is a vital protein for endogenous synthesis of saturated and monounsaturated long-chain fatty acids in some mammals. Nevertheless, its role in lipid synthesis in buffalo mammary gland is still unclear. In this work, the full-length coding sequence (CDS) of ELOVL6 was cloned and identified from buffalo mammary gland. As a result, the CDS of this gene is 795 bp, which encodes a polypeptide of 264 amino acid residues. The buffalo ELOVL6 contains an ELO domain which belongs to the ELO superfamily. Among the 10 tissues of buffalo in peak lactation detected by RT-qPCR, the expression level of ELOVL6 was the highest in the brain, followed by the spleen, and then decreased in the mammary gland, muscle, kidney, heart, liver, rumen, intestine and lung. However, only the expression in the brain and spleen was statistically different from that in other tissues (p < 0.05). Compared with that of the dry-off period, the mRNA abundance of ELOVL6 in the mammary gland was significantly increased in peak lactation. The experiments based on lentivirus transfection in buffalo mammary epithelial cells (BuMECs) displayed that the overexpression of ELOVL6 markedly promoted the expression of INSIG1, INSIG2, SREBP, PPARG, FASN, GPAM, DGAT2 and APGAT6 genes, and the knockdown of ELOVL6 significantly decreased the mRNA abundance of INSIG2, SREBP, FASN, SCD, GPAM, APGAT6 and TIP47 genes. In addition, the increase or decrease of ELOVL6 expression level also caused the corresponding change of total triglyceride content in the BuMECs. The results here suggest that the ELOVL6 can catalyse the synthesis of long-chain fatty acids in the BuMECs, and it can indirectly affect the expression of genes related to milk fat synthesis through its catalytic products to promote the lipid biosynthesis of BuMECs.
最近的研究表明,极长链脂肪酸延长酶6(ELOVL6)是某些哺乳动物内源性合成饱和及单不饱和长链脂肪酸的一种重要蛋白质。然而,其在水牛乳腺脂质合成中的作用仍不清楚。在本研究中,从水牛乳腺中克隆并鉴定了ELOVL6的全长编码序列(CDS)。结果显示,该基因的CDS为795 bp,编码一个由264个氨基酸残基组成的多肽。水牛ELOVL6包含一个属于ELO超家族的ELO结构域。通过RT-qPCR检测泌乳高峰期水牛的10种组织,ELOVL6的表达水平在脑中最高,其次是脾脏,然后在乳腺、肌肉、肾脏、心脏、肝脏、瘤胃、肠道和肺中降低。然而,只有脑和脾中的表达与其他组织存在统计学差异(p < 0.05)。与干奶期相比,泌乳高峰期乳腺中ELOVL6的mRNA丰度显著增加。基于慢病毒转染水牛乳腺上皮细胞(BuMECs)的实验表明,ELOVL6的过表达显著促进了INSIG1、INSIG2、SREBP、PPARG、FASN、GPAM、DGAT2和APGAT6基因的表达,而ELOVL6的敲低显著降低了INSIG2、SREBP、FASN、SCD、GPAM、APGAT6和TIP47基因的mRNA丰度。此外,ELOVL6表达水平的升高或降低也导致了BuMECs中总甘油三酯含量的相应变化。这里的结果表明,ELOVL6可以催化BuMECs中长链脂肪酸的合成,并且它可以通过其催化产物间接影响与乳脂肪合成相关基因的表达,从而促进BuMECs的脂质生物合成。