摘要
目的 探讨糖基化终产物 (AGEs)对内皮细胞过氧化物酶体增生物活化受体 (PPAR)表达的影响。 方法 以体外培养的人内皮细胞系 (ECV) 30 4为研究对象 ,应用RT PCR、Western印迹方法观察不同浓度AGEs对内皮细胞PPARα、PPARγ表达的影响。 结果 ECV 30 4有PPARγ、PPARα的基础表达 ,AGEs可使PPARγ的mRNA及蛋白表达上调。 结论 AGEs对PPAR表达具有上调作用 ,可能参与糖尿病致动脉粥样硬化的病理过程。
Objective To test the hypotheses that human endot helial cells(EC) expresses peroxisome proliferator activated receptor α(PPAR α) and/or PPARγ and that AGEs modulate the expression of PPARα, and/or PPARγ in vitro in a concentration-dependent manner. Methods Human EC (ECV-304 cell line) were cultured in RPMI (Gibco-BRL) supplemented with 10 % FCS(HyClone). Total RNA from EC was isolated by Trizol (Gibco-BRL). The RT -PCR was used to prepare the enouph amount of cDNA of PPARα, PPARγ and GAPDH( Stratagen). For Western blot analysis, the cells were lysed in lysing buffer. Protein concentration was determined by BCA (Pierce). Antigen detection was performed with a chemiluminescent detection system (Pierce). Results Human EC expressed PPARα and PPARγ mRNA and the associated protein. Treatment of EC with the AGEs increased PPARγ mRNA and protein expression in v itro in a concentration-dependent manner. PPARα mRNA and protein expression w as not significantly altered by AGEs. Conclusion AGEs may act in up-regulating potentially proatherosclerotic PPARγ effects in EC via the i ncreased expression of both PPARγ mRNA and protein, possibly by AGE receptor-m ediated pathway.
基金
湖南省教育厅资助项目 ( 4 0 0 JY 0 0C15 4)
