过表达FGF10促进山羊皮下前体脂肪细胞分化

doi:10.11843/j.issn.0366-6964.2019.10.003

摘要/Abstract

摘要： 本研究在构建山羊FGF10过表达腺病毒载体的基础上，阐明过表达FGF10基因对山羊皮下前体脂肪细胞分化的影响及可能的作用机制。本试验利用胶原酶消化法分离获得山羊皮下前体脂肪细胞，采用AdEasy腺病毒包装系统成功构建过表达腺病毒pAdTrack-CMV-FGF10，并感染细胞。采用油红O染色方法从形态学上观察FGF10对成脂分化的影响；利用qPCR技术检测脂肪细胞分化标志基因、脂代谢相关基因、成纤维细胞生长因子受体（fibroblast growth factor receptors，FGFRs）和Kruppel样因子家族（Kruppel like factors，KLFs）mRNA的相对表达变化。结果显示，在过表达FGF10后的第2天，皮下脂肪细胞中脂滴聚集显著多于对照组，C/EBPα、LPL、ACACA、FGFR1和FGFR3的相对表达水平极显著上调（P<0.01），FASN和ATGL的相对表达水平分别出现显著上调（P<0.05）和显著下调（P<0.05），同时，过表达FGF10显著上调KLF家族成员KLF8-10、16和17基因mRNA相对表达水平（P<0.05），显著下调KLF1、2、4和15基因的相对表达水平（P<0.05）。结果表明，过表达FGF10基因可能通过调控C/EBPα、LPL、FASN、ACACA、ATGL和KLFs部分成员的表达促进山羊皮下前体脂肪细胞的分化及脂滴聚集，结果为进一步阐明其调控山羊不同部位脂肪沉积的分子机理提供重要的数据支撑。

Abstract: The aim of this study was to explore the effect and mechanism of FGF10 overexpression on the differentiation of subcutaneous preadipocytes in goats, which was mediated by adenovirus transporters. In this study, the subcutaneous preadipocytes were separated from goat by collagenase digestion, pAdTrack-CMV-FGF10 was constructed successfully through AdEasy adenovirus packaging system, and then was used to infect cells. In morphology, Oil red O staining was used for investigating the effect of FGF10 on adipogenic differentiation. The mRNA levels of adipocyte differentiation marker genes, lipid metabolism related genes, fibroblast growth factor receptors (FGFRs) and Kruppel like factors (KLFs) were measured by quantitative real-time PCR (qPCR). The results showed that the lipid droplets accumulation increased after 2 days of FGF10 overexpression, compared with control group.The mRNA levels of C/EBPα, LPL, ACACA, FGFR1 and FGFR3 significantly increased(P<0.01). The relative expression of FASN and ATGL were significantly up-and down-regulated (P<0.05), respectively. Meanwhile, overexpression of FGF10 significantly up-regulated the mRNA expression of KLF8-10, 16 and 17 genes(P<0.05), while significantly down-regulated the mRNA expression of KLF1, 2, 4 and 15 genes(P<0.05). These results indicate that FGF10 overexpression promotes lipid droplets accumulate possibly by regulating the expression of C/EBPα, LPL, FASN, ACACA, ATGL and partial members of KLFs. This study provides an important data support for elucidating the molecular mechanism for FGF10 controlling fat deposition in different parts of goats.

中图分类号:

S827.2

许晴, 李倩, 王永, 朱江江, 张亚楠, 林亚秋. 过表达FGF10促进山羊皮下前体脂肪细胞分化[J]. 畜牧兽医学报, 2019, 50(10): 1972-1984.

XU Qing, LI Qian, WANG Yong, ZHU Jiangjiang, ZHANG Ya'nan, LIN Yaqiu. Overexpression of FGF10 Promotes the Differentiation of Subcutaneous Preadipocyte in Goat[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 1972-1984.

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