Effects of the Selective EP2 Receptor Agonist Omidenepag on Adipocyte Differentiation in 3T3-L1 Cells
Effects of the Selective EP2 Receptor Agonist Omidenepag on Adipocyte Differentiation in 3T3-L1 Cells
复制标题
DOI：
10.1089/jop.2019.0079
复制
发表时间：
2020-01-14
影响因子：
2.3
通讯作者：
Shams, Naveed K.
中科院分区：
文献类型：
--
作者：
Yamamoto, Yasuko;Taniguchi, Takazumi;Shams, Naveed K.
目的：我们的目的是比较奥美拉唑（OMD）和前列腺素F（FP）受体激动剂（FP激动剂）对小鼠3 T3-L1细胞脂肪生成的影响。研究方法：为了评价OMD对小鼠EP 2（mEP 2）受体的激动活性，我们通过放射免疫测定法测定了表达mEP 2受体的CHO细胞中的cAMP含量。总之，将3 T3-L1细胞在分化培养基中培养10天，并根据油红O染色的细胞面积评估脂肪细胞分化。脂肪形成转录因子Pparg、Cebpa和Cebpb的表达水平的变化通过使用实时聚合酶链反应（PCR）来确定。在脂肪形成过程中，向细胞培养基中加入0.1、1、10和40 μ mol/L的OMD、0.1 μ mol/L的拉坦前列素游离酸（LAT-A）或0.1 μ mol/L的前列腺素F-2 α（PGF（2 α））。将OMD或FP激动剂的油红O染色面积和转录因子靶点的表达模式与未处理的对照进行比较。结果：OMD对mEP 2受体的半数有效浓度（EC 50）为3.9nmol/L。在第10天，在对照细胞内观察到油红O染色的脂滴积聚。LAT-A和PGF（2 α）显著抑制脂滴的积累;然而，OMD即使在高达40 μ mol/L的浓度下也对该过程没有影响。LAT-A和PGF（2 α）在脂肪细胞分化过程中显著抑制Pparg、Cebpa和Cebpb基因表达水平。相反，OMD对这些基因的表达水平没有明显影响。结论：选择性EP 2受体激动剂OMD不影响3 T3-L1细胞的脂肪细胞分化，而FP激动剂显著抑制这一过程。
Purpose: We aimed at comparing the effects of omidenepag (OMD) with those of prostaglandin F (FP) receptor agonists (FP agonists) on adipogenesis in mouse 3T3-L1 cells. Methods: To evaluate the agonistic activities of OMD against the mouse EP2 (mEP2) receptor, we determined cAMP contents in mEP2 receptor-expressing CHO cells by using radioimmunoassays. Overall, 3T3-L1 cells were cultured in differentiation medium for 10 days and adipocyte differentiation was assessed according to Oil Red O-stained cell areas. Changes in expression levels of the adipogenic transcription factors Pparg, Cebpa, and Cebpb were determined by using real-time polymerase chain reaction (PCR). OMD at 0.1, 1, 10, and 40 mu mol/L, latanoprost free acid (LAT-A) at 0.1 mu mol/L, or prostaglandin F-2 alpha (PGF(2 alpha)), at 0.1 mu mol/L were added to cell culture media during adipogenesis. Oil Red O-stained areas and expression patterns of transcription factor targets of OMD or FP agonists were compared with those of untreated controls. Results: The 50% effective concentration (EC50) of OMD against the mEP2 receptor was 3.9 nmol/L. Accumulations of Oil Red O-stained lipid droplets were observed inside control cells on day 10. LAT-A and PGF(2 alpha) significantly inhibited the accumulation of lipid droplets; however, OMD had no effect on this process even at concentrations up to 40 mu mol/L. LAT-A and PGF(2 alpha) significantly suppressed Pparg, Cebpa, and Cebpb gene expression levels during adipocyte differentiation. Conversely, OMD had no obvious effects on the expression levels of these genes. Conclusions: A selective EP2 receptor agonist, OMD, did not affect the adipocyte differentiation in 3T3-L1 cells, whereas FP agonists significantly inhibited this process.