关键词: 游离脂肪酸, 软脂酸, 脐静脉内皮细胞, PTEN, 双过氧钒, 免疫印迹法, 实时定量反转录聚合酶链反应, 人

Abstract: Objective To study the protective effect of PTEN inhibitors on palmitate induced apoptosis human umbilical vein endothelial cell(HUVEC). Methods MTT was used to determine cell survival; Hoechst-PI staining fluorescence microscopy was carried out to observe the morphology of apoptosis; Annexin-Ⅴ-PI flow cytometry was used to determine cell apoptosis; Western blotting was used to analize total PTEN and phosphorylated PTEN protein levels; Real-time quantitative PCR was for the expression of PTEN mRNA. Results MTT results showed palmitic acid(PA) (0.2-0.6mmol/L) for 24 hours significantly inhibited the growth of HUVEC cells; Hoechst-PI staining fluorescence microscopy and Annexin-Ⅴ-PI double staining flow cytometry confirmed that different concentrations of PA (0.2-0.6mmol/L) for 24 hours, with the PA concentration (0.2-0.6mmol/L) increassed, enhanced cell apoptosis; Real-time quantitative PCR and Western blotting showed that, the transcriptional and activity of PTEN increased with the increasing concentration in a dose dependent manner; The role of PIC can increase cell survival, inhibit apoptosis, decrease PTEN transcription and protein expression. Conclusion PA induced apoptosis in HUVEC cells in certain concentration; In the process, PTEN expression and activity increased, PTEN inhibitor bpV reduced cell apoptosis, in the process PTEN transcription and protein expression decreased; PTEN signaling pathway might be an important signal pathway, PTEN inhibitor could inhibit this path and play a protective effect on cells.

Key words: Free fatty acid, Palmitic acid, Umbilical vein endothelial cell, PTEN, Diperoxovahdate, Western blotting, Real-time PCR, Human