화학공학소재연구정보센터
Biochemical and Biophysical Research Communications, Vol.519, No.3, 591-596, 2019
Effects of hypoxia inducible factor-1 alpha on expression levels of MLCK, p-MLC and ZO-1 of rat endothelial cells
Objective: To examine the aberrant expression of endothelial permeability associated proteins including MLCK, p-MLC and ZO-1 in presence of different levels of hypoxia-inducible factor 1 alpha (HIF-1 alpha). Methods: We established monolayer vascular endothelial cell model with the primary rat endothelial cells. Over-expressed or under-expressed HIF-1 alpha cell lines were made by endothelial cells transfected with plasmid vector constructed with HIF-1 alpha gene or HIF-1 alpha-specific short hairpin RNA (shRNA). Levels of mRNA and protein of MLCK, p-MLC and ZO-1 were determined using Real-Time PCR and Western blot. All data were analyzed using by One-Way ANOVA method and LSD. Results: We successfully cultured the rat endothelial primary cells for four days. The mRNA and protein levels of MLCK and p-MLC were significantly increased in the HIF-1 alpha over-expression group than that in the blank control group and the empty plasmid GV230 group (P < 0.05). ZO-1 was significantly lower in the HIF-1 alpha over-expression group than that in the blank control group and the GV230 group. On the contrary, the mRNA and protein levels of MLCK and p-MLC were significantly lower in the HIF-1 alpha under-expression group than that in the blank control group and the shRNA-NC group (P< 0.05). ZO-1 was significantly higher in the HIF-1 alpha low-expression group than that in the blank control group and the shRNA-NC group. Conclusion: HIF-1 alpha positively regulates the expression of MLCK and p-MLC and negatively regulates the expression of ZO-1 in rat monolayer endothelial cells. (C) 2019 Elsevier Inc. All rights reserved.