全反式视黄酸对3T3-L1前脂肪细胞分化及PPARγ和C/EBPα mRNA表达的影响

Effects of all trans retinoic acid on differentiation and expression of PPARγand C/EBPαmRNA in 3T3-L1 preadipocytes

本研究旨在探讨全反式视黄酸(all trans retinoic acid,ATRA)对3T3-L1前脂肪细胞分化的影响及其机制。培养3T3-L1前脂肪细胞,分别设置对照组以及10-9、10-8、10-7、10-6和10-5 mol/L ATRA的处理组,油红O染色观察3T3-L1前脂肪细胞分化的形态变化;油红O染色提取法分析不同浓度ATRA对3T3-L1前脂肪细胞分化的影响;Real-time PCR技术检测脂肪细胞分化相关基因过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptor,PPARγ)、CCAAT/增强子结合蛋白α(CCAAT/enhancer binding proteinα,C/EBPα)mRNA水平的表达。结果表明:10-7、10-6和10-5 mol/L的ATRA抑制3T3-L1前体脂肪细胞的分化(P<0.01),而10-9和10-8 mol/L的ATRA对3T3-L1前体脂肪细胞分化没有显著影响(P>0.05)。10-5 mol/L的ATRA处理的前脂肪细胞,分化相关基因PPARγ和C/EBPαmRNA的表达减少,与空白对照组比较差异有统计学意义(P<0.01)。结果显示,高浓度的ATRA可以抑制3T3-L1前脂肪细胞分化,减少脂肪细胞分化过程中脂质的堆积,机制可能与其抑制脂肪细胞分化相关基因PPARγ和C/EBPαmRNA表达有关。

This study was conducted to explore the effect and mechanism of all trans retinoic acid(ATRA)on differentiation of murine 3T3-L1 preadipocytes.3T3-L1 preadipocytes were cultured and treated with the absence or presence of 10-9,10-8,10-7,10-6 and 10-5 mol/L ATRA.Oil red O staining method were applied to observe morphological changes of the 3T3-L1 cells in differentiating and were eluted with isopropanol to analyze the degree of differentiation.The expressions of peroxisome proliferation activated receptorγ(PPARγ),CAAT/enhancer binding proteinα(C/EBPα)mRNA were detected by real-time PCR.The results showed as followed:10-7,10-6 and10-5 mol/L of ATRA inhibited 3T3-L1 preadipocytes to differentiate into mature adipocytes(P<0.01),10-9 and 10-8 mol/L of ATRA had no significant influence on the differentiation of murine3T3-L1preadipocytes(P>0.05).The preadipocyte treated with 10-5 mol/L of ATRA showed that the expression of differentiation related factor PPARγmRNA as well as the expressions of C/EBPαmRNA significantly lessened,as compared with those in the blank control group,the difference was significant(P < 0.01).In conclusion,high concentrations of ATRA block the differentiation of 3T3-L1 preadipocytes and reduce the accumulation of lipid drops in the adipocyte differentiating process,which may be associated with its effects on decreasing the expressions of adipocyte differentiation related gene PPARγand C/EBPαmRNA.