Chen Xiaobo, Wang Qiang, Zhan Leyun, Shu Aihua
Department of Anesthesiology, Three Gorges University People's Hospital, The First People's Hospital of Yichang, No. 2 Jiefang Road, Yichang, 443000 Hubei China.
Springerplus. 2016 Sep 9;5(1):1518. doi: 10.1186/s40064-016-3067-7. eCollection 2016.
The aim of this study was to investigate the effects of docosahexaenoic acid (DHA) on the generation of angiopoietin-2 (Ang-2) by rat brain microvascular endothelial cells under an oxygen- and glucose-deprivation environment (OGD), and its relationship, if any, with cyclooxygenase 2 (COX-2) expression.
Annexin V and propidium iodide apoptosis assay was used to detect apoptosis. Enzyme linked immunosorbent assay was used to detect Ang-2, vascular endothelial growth factor (VEGF), prostaglandin E2 (PGE2), and prostaglandin I2 (PGI2) content. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect Ang-2 and VEGF mRNA expression. Western blot was used to detect expression of COX-2 protein.
DHA reduced the apoptosis rate (P = 0.026) and decreased the secretion of Ang-2, VEGF, PGE2, and PGI2 (P = 0.006, P = 0.000, P = 0.002, P = 0.004 respectively). The relative expression of Ang2 and Vegf mRNA, as well as COX-2 expression, also decreased (P = 0.000, P = 0.005, P = 0.007 respectively). These effects were antagonized by GW9662 (peroxisome proliferator-activated receptor-γ antagonist). COX-2 protein expression levels were positively correlated with Ang2 and Vegf mRNA expression levels (γ = 0.69, P = 0.038 and γ = 0.76, P = 0.032, respectively). Ang-2 and VEGF mRNA levels were positively correlated with Ang-2 (γ = 0.84, P = 0.012) and VEGF (γ = 0.71, P = 0.036) secretion levels respectively.
DHA reduced apoptosis induced by an OGD environment, thus decreasing Ang-2 and VEGF synthesis. This phenomenon was associated with a decrease in COX-2 protein expression, PGE2 and PGI2 secretion, and generation regulation via intracellular transcriptional pathways.
本研究旨在探讨二十二碳六烯酸（DHA）对氧糖剥夺环境（OGD）下大鼠脑微血管内皮细胞血管生成素-2（Ang-2）生成的影响，以及其与环氧合酶2（COX-2）表达之间的关系（如有）。
采用膜联蛋白V和碘化丙啶凋亡检测法检测细胞凋亡。采用酶联免疫吸附测定法检测Ang-2、血管内皮生长因子（VEGF）、前列腺素E2（PGE2）和前列腺素I2（PGI2）的含量。采用逆转录聚合酶链反应（RT-PCR）检测Ang-2和VEGF mRNA表达。采用蛋白质免疫印迹法检测COX-2蛋白的表达。
DHA降低了细胞凋亡率（P = 0.026），并减少了Ang-2、VEGF、PGE2和PGI2的分泌（分别为P = 0.006、P = 0.000、P = 0.002、P = 0.004）。Ang2和Vegf mRNA的相对表达以及COX-2的表达也降低（分别为P = 0.000、P = 0.005、P = 0.007）。这些作用被GW9662（过氧化物酶体增殖物激活受体-γ拮抗剂）拮抗。COX-2蛋白表达水平与Ang2和Vegf mRNA表达水平呈正相关（分别为γ = 0.69，P = 0.038和γ = 0.76，P = 0.032）。Ang-2和VEGF mRNA水平分别与Ang-2（γ = 0.84，P = 0.012）和VEGF（γ = 0.71，P = 0.036）分泌水平呈正相关。
DHA减少了OGD环境诱导的细胞凋亡，从而降低了Ang-2和VEGF的合成。这种现象与COX-2蛋白表达、PGE2和PGI2分泌的减少以及通过细胞内转录途径的生成调节有关。
