PAQR9对棕色脂肪产热及UCP1表达的影响

目的 初步探究孕激素和脂联素受体家族成员9（progestin and adipoQ receptor family member 9，PAQR9）对棕色脂肪产热及解偶联蛋白1（uncoupling protein 1，UCP1）表达的影响。方法 选取C57BL/6J小鼠的白色脂肪组织（white adipose tissue，WAT）、棕色脂肪组织（brown adipose tissue，BAT）及前棕色脂肪细胞系DE2-3和间充质干细胞C3H10T1/2细胞系进行研究。实时荧光定量PCR技术检测不同条件下PAQR9及UCP1的mRNA水平，蛋白免疫印迹法检测UCP1的蛋白质水平。实时荧光定量PCR技术和油红O染色法检测脂肪细胞的成脂分化情况。结果 C57BL/6J小鼠BAT中PAQR9的mRNA水平显著高于WAT。冷刺激诱导下，BAT和腹股沟皮下WAT中PAQR9的mRNA水平显著升高。DE2-3细胞诱导分化成熟即第6天时，PAQR9的mRNA水平显著高表达。在DE2-3细胞中过表达PAQR9，UCP1的mRNA水平显著上调，细胞耗氧量显著增加，而DE2-3细胞成脂分化功能无显著性差异。在DE2-3细胞中敲低PAQR9，UCP1的mRNA水平显著下调。在C3H10T1/2细胞第4~6天脂肪细胞分化成熟时，PAQR9的mRNA水平显著上调，在敲低PAQR9后UCP1蛋白质水平下调，而C3H10T1/2细胞成脂分化功能无显著性差异。结论 PAQR9能够促进棕色脂肪细胞产热，调控产热基因UCP1的表达。

Effects of PAQR9 on brown fat thermogenesis and UCP1 expression

Objective To preliminarily explore the effects of progestin and adipoQ receptor family member 9(PAQR9)on brown fat thermogenesis and the expression of uncoupling protein 1(UCP1). Methods The white adipose tissue (WAT) and brown adipose tissue (BAT) of C57BL/6J mice,pre-brown fat cell line DE2-3 and C3H10T1/2 mesenchymal stem cells were used for studies.Quantitative real-time PCR was used to detect the mRNA levels of PAQR9 and UCP1 under different conditions,and Western blot was used to detect the protein level of UCP1.Quantitative real-time PCR and oil-red O staining were used to detect the adipogenesis of adipocytes. Results The mRNA level of PAQR9 in BAT of C57BL/6J mice was significantly higher than that in WAT.Under cold exposure,the mRNA level of PAQR9 in BAT and inguinal WAT increased significantly.When DE2-3 cells were induced to differentiate on day 6,the mRNA level of PAQR9 was significantly induced.PAQR9 was overexpressed in DE2-3 cells,the mRNA level of UCP1 was up-regulated,and cell oxygen consumption increased significantly,but there was no significant difference in the adipogenesis function of DE2-3 cells. The mRNA level of UCP1 was significantly down-regulated after knocking down PAQR9 in DE2-3 cells.When C3H10T1/2 cells were induced to differentiate and mature on the 4^th-6^th day,the mRNA level of PAQR9 was up-regulated significantly,and the protein level of UCP1 was down-regulated after PAQR9 was knocked down,but there was no significant difference in the adipogenesis function of C3H10T1/2 cells. Conclusion PAQR9 played an important role in activating thermogenesis of brown adipocyte,and regulated the expression of thermogenic gene UCP1.