Molecular characterization of fibroblast growth factor-16 and its role in promoting the differentiation of intramuscular preadipocytes in goat
Molecular characterization of fibroblast growth factor-16 and its role in promoting the differentiation of intramuscular preadipocytes in goat
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DOI：
10.1017/s1751731120001160
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发表时间：
2020-11-01
期刊：
影响因子：
3.6
通讯作者：
Wang, Y.
中科院分区：
文献类型：
--
作者：
Huang, K.;Liang, J. J.;Wang, Y.
脂肪代谢是一个重要而复杂的体内生化反应，受多种因素的调控.近年来，成纤维细胞生长因子-16（FGF-16）在棕色脂肪组织中的高表达引起了人们对探索其在脂肪生成和脂质代谢中的作用的兴趣。本研究克隆了山羊FGF 16基因624 bp，包括完整的开放阅读框架，编码207个氨基酸。我们发现FGF 16在山羊肾脏和心脏中表达最高，其次是皮下脂肪和三头肌。FGF 16在脂肪细胞分化的第2天表达最高（P< 0.01），随后明显下降。本研究采用过表达和干扰的方法研究了山羊肌内前脂肪细胞中FGF 16基因的功能。沉默FGF 16可减少脂肪细胞脂滴聚集和甘油三酯合成。这与FGF 16过表达的情况相反。此外，FGF 16基因敲低还导致CCAAT增强子结合蛋白β（P< 0.01）、脂肪酸结合蛋白-2（P<0.01）和固醇调节元件结合蛋白-1（P< 0.05）等脂肪细胞分化相关基因表达下调，而前脂肪细胞因子-1表达上调。同时，与甘油三酯分解相关的脂肪甘油三酯脂肪酶（P< 0.01）和脂肪敏感脂肪酶（P< 0.05）基因也高表达。接下来，我们通过蛋白质相互作用网络锁定成纤维细胞生长因子受体4（FGFR 4），并干扰FGF 16以显著降低FGFR 4的表达。发现FGFR 4在脂肪细胞分化中的表达谱与FGF 16的表达谱高度相似。过表达和干扰实验证实FGFR 4和FGF 16具有相同的促进脂肪细胞分化的功能。最后，采用共转染技术，将pc-FGF 16和siRNA-FGFR 4、siRNA 2-FGF 16和siRNA-FGFR 4分别联合转染脂肪细胞。结果发现，在FGF 16过表达的情况下，细胞的脂质分泌和甘油三酯合成随干扰浓度的增加呈现先增加后减少的趋势。在干扰FGF 16的情况下，随着干扰浓度的增加，脂质分泌和甘油三酯合成呈下降趋势。这些结果表明，FGF 16通过受体FGFR 4的表达介导脂肪细胞分化，为进一步研究FGF 16在山羊脂肪形成中的功能作用奠定了基础。
Fat metabolism is an important and complex biochemical reactionin vivoand is regulated by many factors. Recently, the findings on high expression of fibroblast growth factor-16 (FGF16) in brown adipose tissue have led to an interest in exploring its role in lipogenesis and lipid metabolism. The study cloned the goat'sFGF16 gene 624 bp long, including the complete open reading frame that encodes 207 amino acids. We found thatFGF16 expression is highest in goat kidneys and hearts, followed by subcutaneous fat and triceps. Moreover, the expression ofFGF16 reached its peak on the 2nd day of adipocyte differentiation (P< 0.01) and then decreased significantly. We used overexpression and interference to study the function ofFGF16 gene in goat intramuscular preadipocytes. Silencing ofFGF16 decreased adipocytes lipid droplet aggregation and triglyceride synthesis. This is in contrast to the situation whereFGF16 is overexpressed. Furthermore, knockdown ofFGF16 also caused down-regulated expression of genes associated with adipocyte differentiation including CCAAT enhancer-binding protein beta (P< 0.01), fatty acid-binding protein-2 (P< 0.01) and sterol regulatory element binding protein-1 (P< 0.05), but the preadipocyte factor-1 was up-regulated. At the same time, the genes adipose triglyceride lipase (P< 0.01) and hormone-sensitive lipase (P< 0.05) associated with triglyceride breakdown were highly expressed. Next, we locked the fibroblast growth factor receptor-4 (FGFR4) through the protein interaction network and interfering withFGF16 to significantly reduceFGFR4 expression. It was found that the expression profile ofFGFR4 in adipocyte differentiation was highly similar to that ofFGF16. Overexpression and interference methods confirmed thatFGFR4 andFGF16 have the same promoting function in adipocyte differentiation. Finally, using co-transfection technology, pc-FGF16 and siRNA-FGFR4, siRNA2-FGF16 and siRNA-FGFR4 were combined to treat adipocytes separately. It was found that in the case of overexpression ofFGF16, cell lipid secretion and triglyceride synthesis showed a trend of first increase and then decrease with increasing interference concentration. In the case of interference withFGF16, lipid secretion and triglyceride synthesis showed a downward trend with the increase of interference concentration. These findings illustrated thatFGF16 mediates adipocyte differentiation via receptorFGFR4 expression and contributed to further study of the functional role ofFGF16 in goat fat formation.