NR4A1对小鼠卵巢颗粒细胞脂肪代谢相关调控因子的调节作用

目的观察NR4A1对小鼠卵巢颗粒细胞中脂肪代谢相关调控因子脂联素受体2（Adipo—R2）、解偶联蛋白2（UCP2）、B类清道夫受体（CD36）的调节作用。方法分别构建NR4A1超表达腺病毒载体（AdCMV～NR4A1）及干涉腺病毒载体（AdHl－siRNA／NR4A1）。体外培养小鼠卵巢颗粒细胞，贴壁生长至90％时，分为病毒感染组、胰岛素（100nmol／L）与胰岛素增敏剂（10μmol／L）组和病毒＋胰岛素组。Trizol裂解细胞提取总RNA，实时荧光定量PCR法检测颗粒细胞中Adipo—R2，UCP2，CD36的mRNA表达水平。结果在小鼠卵巢颗粒细胞中超表达NR4A1能够促进Adipo—R2，UCP2，CD36基因的转录（P〈O．05），RNA干扰NR4AI后则可以抑制以上基因的表达（P〈0．05）；胰岛素可以促进UCP2，CD36基因的表达（P〈O．05），而对Adipo—R2的作用不明显；另外胰岛素增敏剂曲格列酮也能够促进Adipo—R2，UCP2，CD36的表达（P〈0．05）；RNA干扰NR4A124h后，再向细胞中加入生理剂量（100nmol／L）胰岛素30rain后Adipo—R2，UCP2，CD36的表达水平与单独干涉组比均可逆转升高（P〈O．05），其中UCP2的表达水平可升高到胰岛素组同样的水平，但Adipo—R2，CD36的逆转升高水平并没有达到仅加入胰岛素时的升高水平（P〈O．05）。结论NR4A1对Adipo—R2，UCP2，CD36具有正向调节作用，同时与胰岛素或胰岛素增敏剂可能有协同调节作用，进而发挥对脂肪代谢的调控。

NR4A1 orphan nuclear receptors regulated the expression of lipid metabolism-related factors in mouse granulosa cells

Objective. To examine the effect of NR4A1 on the regulation of three lipid metabolism-related factors, adiponeetin receptor 2 （Adipo-R2）, uncoupling protein 2 （UCP2） and scavenger receptor B （CD36） in mouse granulosa cells. Methods： Recombinant adenovirus AdCMV-NR4A1 and AdHI-siRNA/NR4A1 were constructed to enhance and knock down the expression of NR4A1, respectively. Mouse granulosa cells were isolated and cultured in vitro to reach a confluency of 90% or more. Cultured cells were infected with recombinant adenovirus, or treated with insulin （100 nmol/L） or troglitazone （10 μmol/L）, or infected with recombinant adenovirus plus insulin treatment. Total RNA of the cultured cells were extracted by Trizol after the treatment and the mRNA expressions of Adipo-R2, UCP2, CD36 were detected by Real-time polymerase chain reaction （PCR）.Results： The mRNA expressions of Adipo-R2, UCP2, CD36 were significantly increased in mouse granulosa cells infected with AdCMV-NR4A1 （P〈0.05）, while significantly inhibited in those infected with AdHI-siRNA/NR4A1 （P〈0.05） ; Troglitazone increased the mRNA expressions of all three studied factors in the cultured cells （P〈0.05） ; Expressions of UCP2 and CD36 were up-regulated by insulin, but Adipo-R2 expression was unchanged; The expressions of Adipo-R2, UCP2, CD36 in the cells infected with AdHI-siRNA/NR4A1 were significantly increased by insulin stimulation, when compared with those in the cells infected with AdHI-siRNA/NR4A1 alone （P〈0.05）, and Adipo-R2 expression could reach the same levels as in the cells treated with insulin alone. Conclusions： NR4A1 could up-regulate the expressions of Adipo-R2, UCP2 and CD36 in cultured mouse granulose cells, and may have a synergistic effect with insulin or troglitazone.