共轭亚油酸是具有一定功能的多不饱和脂肪酸，但反式脂肪酸的功能至今仍有争议，本研究拟探讨实验室自制的t9，t11-CLA对氧化损伤的人脐静脉内皮细胞（HUVECs）的保护作用并对其可能机理进行解析。采用MTT法分析t9，t11-CLA对氧化损伤HUVECs的保护作用，通过流式细胞术检测t9，t11-CLA对细胞凋亡及线粒体膜电位变化的影响，采用实时荧光定量PCR和caspase蛋白试剂盒检测与细胞凋亡相关的caspase蛋白及基因变化情况。结果表明，t9， t11-CLA能显著增强HUVECs抵御氧化损伤的能力，缓解氧化损伤造成的细胞膜电位下降、细胞凋亡比例上升、caspase-8、-3活性激活及caspase-8、-3 mRNA表达量的上调，说明t9，t11-CLA可能通过与线粒体途径相关的caspase依赖的抗凋亡机制保护了血管内皮细胞免受氧化损伤。%Conjugated linoleic acids (CLA) have been extensively studied in both in vivo and in vitro models because of their beneficial biological activities. However, the effect of trans CLA were controversial. Thus, the aim of the study was to explore the protective effects and the potential mechanism of t9,t11-CLA on the oxidative damage of human umbilical vein endothelial cells (HUVECs). The oxidative injury model was established with H2O2. MTT assay was used to assess inhibitory effect of t9,t11-CLA on HUVECs. Apoptosis and mitochondrial membrane potential (ΔΨm) measurements were analyzed using flow cytometry. The mRNA expression and activities of caspases-3 and-8 were determined using caspase assay kits and real time quantitative PCR. Results showed that t9,t11-CLA protected HUVECs against oxidative injury. Compared with the H2O2 group, increases in ΔΨm, decreases in the rate of apoptosis and activities, mRNA expression of caspase-3and-8 were clearly observed upon exposure to different t9,t11-CLA concentrations, which suggested that the protective mechanism of t9,t11-CLA HUVECs against oxidative injury involves the caspase-dependent mitochondrial apoptosis pathway.
展开▼
