脂肪酸对延边牛前体脂肪细胞UCP1、UCP3的mRNA表达的影响

Effects of Fatty Acids on The mRNA Expression of UCP1 and UCP3 in Yanbian Cattle Preadipocytes

试验旨在探索前体脂肪细胞受到冷应激后的响应机制,并初步鉴定以单不饱和脂肪酸中的油酸(OA)、肉豆蔻油酸(MOA)对解偶联蛋白1(UCP1)、解偶联蛋白3(UCP3)基因表达的影响,探索以OA、MOA为代表的单不饱和脂肪酸与细胞冷应激之间是否存在关联。以OA、MOA以及延边牛前体脂肪细胞作为试验材料,分别用OA、MOA处理前体脂肪细胞96 h,通过油红O染色和三酰甘油含量的测定来评估前体脂肪细胞内脂肪生成情况,利用qPCR测定UCP1、UCP3基因的相对表达量。再分别对未经处理、OA处理96 h、MOA处理96 h的前体脂肪细胞进行4℃冷应激处理0、6、12、24 h后,分别检测其UCP1、UCP3基因的相对表达量。结果显示:OA组与MOA组均可显著降低UCP1和UCP3基因的表达水平(P<0.05);在冷处理6~24 h后UCP1基因表达量发生显著性上升(P<0.05),冷处理6~12 h的UCP3基因表达量发生显著性降低(P<0.05)。试验发现,OA、MOA处理前体脂肪细胞96 h后,再进行4℃冷应激处理24 h,会导致UCP1基因表达量上升,而UCP3的表达量随着冷应激处理时间的延长而降低,并且试验初步验证了OA及MOA对两种基因具有抑制作用。

The purpose of the experiment was to explore the response mechanism of preadipocytes after cold stress,and preliminarily identify the effect of oleic acid(OA)and myristic acid(MOA)in monounsaturated fatty acids on the expression of UCP1 and UCP3,and explore whether there was a relationship between monounsaturated fatty acids represented by OA and MOA and cold stress in cells.Oleic acid OA,myristic acid MOA,and Yanbian cattle preadipocytes were used as test materials.The preadipocytes were treated with OA and MOA for 96 h respectively,and the fat production in preadipocytes was determined by oil red staining and triglyceride content,and the relative surface amount of UCP1 and UCP3 was determined by qPCR.At the same time,the relative expression of UCP1 and UCP3 were measured after the adipocytes were treated with cold stress(4℃)for 0 h,6 h,12 h and 24 h before the treatment of untreated,OA and MOA for 96 h.The results showed that the expression levels of UCP1 and UCP3 were significantly decreased in OA group and MOA group(P<0.05);the expression of UCP1 increased significantly(P<0.05).The experiment found that the expression of UCP1 could be activated when the preadipocytes cultured for 96 h in 4℃cold environment for 24 h,and the expression of UCP3 decreased with the time of cold stress treatment.The experiment preliminarily verify the inhibition of oleic acid and myristic acid on the two genes,providing basic data for the subsequent control of the application of molecular biological technology related to cold-resistant genes in animal husbandry.