ASIP基因及其重组蛋白对山羊脂肪细胞脂肪沉积的效应

Effect of ASIP Gene and Recombinant ASIP Protein on Lipid Deposition in Goat Adipocytes in Vitro

为探索ASIP基因对山羊脂肪沉积的效应,本研究采集波杂山羊(BZ)和酉州乌羊(YZ)组织,并使用体外培养的山羊前脂肪细胞,采用基因克隆、实时荧光定量(qRT-PCR)、油红O染色等方法,检测ASIP基因在山羊组织和体外培养分化脂肪细胞中的表达规律,以及脂质生成关键基因在山羊脂肪和肌肉组织中的表达情况,分析重组ASIP蛋白对山羊脂肪细胞分化过程中脂滴形成及脂质生成关键基因表达的影响。结果表明,ASIP基因在山羊体组织中广泛表达,在肾脏和白色脂肪中的表达量最高;ASIP基因在山羊脂肪细胞分化6 d时显著升高(P<0.05),8 d时达到最高值。PPARG、LEPTIN和ATRN基因在YZ肾脂中的表达量显著高于BZ(P<0.05);PPARG、FASN和ATRN基因在BZ骨骼肌中的表达量显著高于YZ(P<0.05),FABP4和SREBP1基因的表达在两品种羊之间无显著差异。重组ASIP蛋白处理诱导山羊脂肪细胞在8 d出现明显脂肪滴,显著提高了细胞内甘油三酯含量(TG)(P<0.05),并显著或极显著上调脂肪细胞中脂质合成关键基因(PPARG、LEPTIN、FASN、FABP4、SREBP1和ATRN)的表达量。综上,ASIP基因及重组ASIP蛋白对山羊脂肪细胞的脂肪沉积可能具有正向调控作用。本研究结果为进一步解析ASIP基因调控山羊脂肪沉积机制提供了重要理论基础。

To explore the effect of the ASIP gene and recombinant ASIP protein on lipid deposition in goat,gene cloning,quantitative real-time PCR(qRT-PCR),and Oil-Red-O staining were used as methods,tissues and cultured adipocytes of Boer crossed goat(BZ)and Youzhou dark goat(YZ)were taken as materials.The expression pattern of the ASIP gene in both tissues and differentiated adipocytes was characterized,and the expression levels of key lipogenesis genes in adipose and muscle tissues in BZ and YZ were also detected.Furthermore,the effects of recombinant ASIP protein on lipid droplet formation and the expression levels of lipogenesis genes during goat adipocyte differentiation were examined.The results showed that the ASIP gene was widely expressed in goat tissues,with the highest expression levels in kidneys and white adipose tissue.The expression level of the ASIP gene significantly increased on day 6 of goat adipocyte differentiation(P<0.05)and reached the highest on day 8(P<0.05).The expression levels of PPARG,LEPTIN,and ATRN genes in YZ adipose were significantly higher than those in BZ(P<0.05).The expression level of PPARG,FASN,and ATRN genes in BZ muscle was significantly higher than those in YZ(P<0.05).While the expression levels of FABP4 and SREBP1 genes did not show a significant difference between YZ and BZ.Recombinant ASIP protein evidently induced the formation of fat droplets during adipocyte differentiation on day 8,significantly increased the content of intracellular triglyceride(TG)(P<0.05),and significantly upregulated the expression levels of essential lipogenesis genes(PPARG,LEPTIN,FASN,FABP4,SREBP1,ATRN)in goat adipocytes.In conclusion,ASIP gene and the recombinant ASIP protein probably have a positive effects on goat fat deposition.This study provides a theoretical basis for further revealing the mechanisms by which the ASIP gene influences fat deposition in goat.