Abstract

This study aimed to clone the full-length open reading frame (ORF) of goat ANGPTL8 gene sequence, reveal its molecular and expression characteristics, and explore its effect on the differentiation of goat intramuscular preadipocytes. The full-length ORF sequence of goat ANGPTL8 gene was cloned by RT-PCR technology, and bioinformatics analysis was performed by related biological software. RT-qPCR was used to detect the expression of ANGPTL8 mRNA in goat tissues. Further use of RNA interference to study the effect of ANGPTL8 on the differentiation of goat intramuscular preadipocytes. The total length of the ANGPTL8 gene nucleotide sequence is 717 bp, including 597 bp of ORF, encoding 198 amino acids. Goat ANGPTL8 has the closest relationship with sheep, it was widely expressed in different tissues, and relatively enriched in liver. The silence of ANGPTL8 inhibited the accumulation of lipid droplets by 5.76% in goat intramuscular preadipocytes (p > 0.05) and significantly suppressed the expression of the genes related to preadipocytes differentiation, fatty acid synthesis and transport (p＜0.05 or p＜0.01). These data illuminate the speculation that ANGPTL8 may involve in the lipid accumulation regulation via the control of PPARγ and C/EBPβ in goat adipocytes.

摘要

本研究旨在克隆山羊ANGPTL8基因序列的全长开放阅读框（ORF），揭示其分子和表达特征，探讨其对山羊肌内前脂肪细胞分化的影响。采用RT-PCR技术克隆山羊ANGPTL8基因全长ORF序列，并通过相关生物学软件进行生物信息学分析。RT-qPCR用于检测山羊组织中ANGPTL8 mRNA的表达。进一步利用RNA干扰研究ANGPTL8对山羊肌内前脂肪细胞分化的影响。ANGPTL8基因核苷酸序列全长717 bp，其中ORF 597 bp，编码198个氨基酸。山羊ANGPTL8与绵羊的关系最密切，在不同组织中广泛表达，在肝脏中相对富集。ANGPTL8的沉默抑制了山羊肌肉前脂肪细胞中脂滴积累5.76%（p  >0.05），并显着抑制了前脂肪细胞分化、脂肪酸合成和转运相关基因的表达（p <0.05或p <0.01）。这些数据阐明了ANGPTL8可能通过控制山羊脂肪细胞中的PPARγ和C/EBPβ参与脂质积累调节的推测。