PAEs对原代培养的大鼠脂肪细胞PPAR-γ2表达的影响

Effects of PAEs on the gene expression of PPAR-γ 2 in primary rat adipocytes

摘要:

目的：观察邻苯二甲酸酯类化合物（Phthalate acid esters，PAEs）对原代培养的大鼠脂肪细胞过氧化物酶体增殖剂激活受体-γ 2（Peroxisome prolifcrator activated receptor gamma 2，PPAR-γ 2）mRNA表达的影响。方法：对脂肪细胞进行PAEs染毒培养，观察脂肪细胞多方面的变化，评价PAEs对脂肪细胞的影响。主要实验：①显微镜每天观察脂肪细胞的形态变化；②油红O染色提取法和MTT比色法检查脂肪细胞的生长状况；③提取脂肪细胞的总RNA，通过逆转录PCR实验，分析PPAR-γ 2 mR－NA的表达情况。结果：①PAEs处理过的脂肪细胞由最初的小圆形变化为梭形最后变成椭圆形，大鼠脂肪细胞内出现脂肪颗粒，并逐渐增加，最后部分脂肪颗粒融合为较大的脂肪颗粒，符合脂肪细胞的发育规律。②随着培养液中PAEs浓度的增加，大鼠脂肪细胞产生的脂肪颗粒数量增多。③PPAR-γ 2的表达随着PAEs浓度的增加而增加。结论：可以认为PAEs通过增强PPAR-γ 2表达来促进脂肪细胞的生长发育，研究结果将会为研究PAEs与肥胖的关系提供科学依据，为进一步研究PAEs对健康的影响提供了一个新的思路。

Abstract:

Objective：To observe the effects of phthalate acid esters （PAEs） on the gene expression of peroxisome prolifcrator activat－ed receptor gamma2 （PPAR-γ2） in primary rat adipocytes. Methods：Adipocytes in cultures were exposed to PAEs. Main experi－ments：①observed morphological changes of adipocytes by microscope every day；②Oil red O staining and MTT assay method were used to check the growth status of adipocytes；③Total RNA from these adipocytes was extracted by reverse transcription PCR. Results：①Morphological of adipocytes with PAEs changed from Small round to spindle and finally turned into oval. lipochondrion was observed in adipocytes. Along with the increase of culturing time，lipochondrion in adipocytes gradually increase，finally part of lipochondrion fusion to a bigger lipochondrion，accord with the developing law of adipocytes.②A large lipochondrion was present as medium of concentration of PAEs increased；③The expression of PPAR-γ2 increased as concentration of PAEs increased. The results of this study will provide the scientific basis for the relationship with obesity，and provide a new idea for the the further research of the effects of PAEs to health. Conclusion：The major role of adipocytes growth perhaps is that PAEs enhanced the expression of PPAR-γ 2.