猪OLR1基因对肌内前脂肪细胞分化的影响

研究氧化型低密度脂蛋白受体1(oxidized low-density lipoprotein receptor1, OLR1)基因在猪前脂肪细胞分化过程中的作用,为深入研究OLR1基因在脂肪沉积中的功能奠定基础。根据GenBank中猪OLR1基因序列(登录号:NM_213805),构建OLR1基因过表达载体OLR1-pcDNA3.1,同时合成干扰OLR1表达的siRNA。从恩施黑猪背最长肌中分离前脂肪细胞,分别将OLR1-pcDNA3.1和OLR1-siRNA转染前脂肪细胞,并对细胞进行诱导分化,至第6天,通过油红O染色、甘油三酯含量测定和荧光定量PCR检测OLR1基因对前脂肪细胞分化的影响。结果显示,OLR1过表达组脂滴明显多于对照组,甘油三酯含量显著(P<0.05)增加,成脂分化标志基因PPARγ的表达量极显著(P<0.01)升高、C/EBPα和FAS的表达量显著(P<0.05)升高;干扰OLR1后,细胞内脂滴减少,甘油三酯含量显著(P<0.05)下降,成脂分化标志基因PPARγ的表达量极显著(P<0.01)下降,C/EBPα和FAS的表达量显著(P<0.05)降低。综上表明,OLR1基因具有促进前脂肪细胞成脂分化的作用。

Effect of porcine OLR1 gene on intramuscular preadipocytes differentiation

The aim of this study was to investigate the role of oxidized low density lipoprotein receptor 1 (OLR1) gene in the differentiation of porcine preadipocytes, and to lay a foundation for further study of the function of OLR1 gene in fat deposition. According to the sequence of OLR1 gene in GenBank (GenBank No.: NM_213805), the over expression vector of OLR1-pcDNA3.1 was constructed and OLR1-siRNA was synthesized. Preadipocytes were isolated from the longissimus dorsi muscle of Enshi black pig. OLR1-pcDNA3.1 and OLR1-siRNA were transferred into the preadipocytes respectively. The cells were induced to differentiate to the 6th day. Then, oil red O staining, triglyceride content detection and fluorescence quantitative PCR were used to detect the effect of OLR1 gene on the preadipocyte differentiation. The results showed that the number of lipid droplets in the overexpression group was significantly(P<0.05) higher than that in the control group, the triglyceride content was significantly increased (P<0.05), and the expression levels of PPARγ, C/EBPα and FAS were significantly increased at P<0.01, P<0.05 and P<0.05 level, respectively. In contrast, after knock down OLR1 gene, the intracellular lipid droplets and triglyceride content were significantly decreased (P<0.05), and the expression of adipogenic differentiation marker genes PPARγ, C/EBPα and FAS were significantly decreased at P<0.01, P<0.05 and P<0.05 level, respectively. In conclusion, OLR1 gene could promote the adipogenic differentiation of preadipocyte.