急性寒冷刺激诱导小鼠米黄色脂肪细胞形成方法的建立

Establishment of the method of acute cold stimulus inducing mouse subcutaneous beige adipogenesis

目的:建立急性寒冷刺激诱导小鼠皮下米黄色脂肪细胞形成的方法。方法:连续3天每天4 h预冷刺激使小鼠适应寒冷环境,然后持续24 h进行急性寒冷刺激,诱导小鼠皮下脂肪中米黄色脂肪细胞形成。采用免疫组化检测皮下脂肪中Ucp1阳性细胞,荧光定量PCR检测米黄色脂肪细胞标志性基因Ucp1、Prdm16、Pgc1α、Cidea的转录,Western blot检测米黄色脂肪细胞标志性蛋白Prdm16和Ucp1的表达。结果:急性寒冷刺激后小鼠皮下脂肪中Ucp1阳性细胞数量明显增加,Ucp1、Prdm16、Pgc1α、Cidea转录明显上调,Prdm16、Ucp1蛋白表达明显升高。结论:本研究建立的急性寒冷刺激诱导小鼠皮下米黄色脂肪细胞形成的方法可靠,可用于米黄色脂肪细胞形成调控机制的研究。

Objective: To establish the method of acute cold stimulus inducing beige adipogenesis in mouse subcutaneous fat. Methods: After 3 times 4℃ stimulus for 4 heachday, the mice were administrated to persistent cold stimulus for 24 hto induce beige adipogenesis in subcutaneous fat. IHC was used to detect the formation of Ucp1 positive adipocytes. qPCR and western blot was performed to monitor the expression of beige adipocytes markers Ucp1, Prdm16, Pgc1αand Cidea. Results: Ucp1 positive adipocytes increased after acute cold stimulus. Transcription of beige adipocytes marker genes Ucp1, Prdm16, Pgc1α and Cidea were upregulated and western blot showed that expression of Ucp1 and Prdm16 increased significantly. Conclusions: The method of beige adipogenesis in mouse subcutaneous fat was established successfully and this inducing method is applicable for the mechanism study of beige adipocytes formation.