Wei Yang1, Yuan Zeng2, Bin Li3, Jianliang Zhou1, Yi Gong1, Jianjun Xu1, Xiao Dong1. 1. Department of Cardiothoracic Surgery, The Second Affiliated Hospital of Nanchang University Nanchang 330008, China. 2. Department of Thoracic Surgery, The First Affiliated Hospital of Guangzhou Medical University Guangzhou 5101203, China. 3. Department of Heart Surgery, Beijing Anzhen Hospital of Capital Medical University Beijing 100029, China.
Abstract
OBJECTIVE: This study aims to explore the relationship between PBEF and VEGF and p-MLC and the mechanism of PBEF increasing permeability of endothelial cells in hypoxia/re-oxygenation. METHODS: Hypoxia/re-oxygenation model was established and PBEF siRNA was synthesized. According to the different HUVEC treatment, it can be divided into normal control group, PBEF siRNA group; hypoxia (20 hours) and re-oxygenation (3 h) group, hypoxia (20 h) and re-oxygenation (6 h) group, hypoxia (20 h) and re-oxygenation (9 hours) group, hypoxia (20 h)/re-oxygenation (12 h). The expressions of PBEF, VEGF and p-MLC were tested by RT-PCR and Western blot. RESULTS: The mRNA and protein expression of PBEF in PBEF siRNA group were significantly lower compared to liposome group and the negative controls (P < 0.05). The expression of PBEF protein in hypoxia/re-oxygenation group was significantly higher than the normal control group. It increased in the 3 h of re-oxygenation group, peaked at 9 h, until 12 h started to decline (P < 0.05). When the PBEF gene was knockdown, the expression of VEGF and p-MLC in hypoxia and re-oxygenation are significantly lower. CONCLUSIONS: PBEF siRNA can effectively inhibit the expression of PBEF in endothelial cells. The expression of PBEF, VEGF and p-MLC were significantly higher in endothelial cell after Hypoxia/re-oxygenation. PBEF may change the permeability of endothelial cells by regulating the expression of VEGF and the phosphorylation of MLC.
OBJECTIVE: This study aims to explore the relationship between PBEF and VEGF and p-MLC and the mechanism of PBEF increasing permeability of endothelial cells in hypoxia/re-oxygenation. METHODS:Hypoxia/re-oxygenation model was established and PBEF siRNA was synthesized. According to the different HUVEC treatment, it can be divided into normal control group, PBEF siRNA group; hypoxia (20 hours) and re-oxygenation (3 h) group, hypoxia (20 h) and re-oxygenation (6 h) group, hypoxia (20 h) and re-oxygenation (9 hours) group, hypoxia (20 h)/re-oxygenation (12 h). The expressions of PBEF, VEGF and p-MLC were tested by RT-PCR and Western blot. RESULTS: The mRNA and protein expression of PBEF in PBEF siRNA group were significantly lower compared to liposome group and the negative controls (P < 0.05). The expression of PBEF protein in hypoxia/re-oxygenation group was significantly higher than the normal control group. It increased in the 3 h of re-oxygenation group, peaked at 9 h, until 12 h started to decline (P < 0.05). When the PBEF gene was knockdown, the expression of VEGF and p-MLC in hypoxia and re-oxygenation are significantly lower. CONCLUSIONS:PBEF siRNA can effectively inhibit the expression of PBEF in endothelial cells. The expression of PBEF, VEGF and p-MLC were significantly higher in endothelial cell after Hypoxia/re-oxygenation. PBEF may change the permeability of endothelial cells by regulating the expression of VEGF and the phosphorylation of MLC.
Authors: Shui Q Ye; Li Q Zhang; Djanybek Adyshev; Peter V Usatyuk; Alexander N Garcia; Tera L Lavoie; Alexander D Verin; Viswanathan Natarajan; Joe G N Garcia Journal: Microvasc Res Date: 2005-09-26 Impact factor: 3.514