JIN Hongguo,ZHOU Min,JIN Meiyu, et al. Study on the expression of RhoA/Rho-kinase and eNOS in vascular endothelial cells under hypoxic condition[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2016, 46(5): 321-325.
Abstract：Objective: To establish stable hypoxic model in vitro and to explore the effect of RhoA protein/Rho-kinase in human umbilical vein endothelial cell, (HUVEC) on eNOS expression of its endothelial cell under hypoxic condition. Methods: SH-SY5Y cells, HEK293 cells and HUVEC were respectively transfected using jet-PEI-HUVEC and siRNA to prepare cellular hypoxic model, the Western blotting analysis was preformed for the related proteins through cellular lysis. Results: RhoA protein levels in HUVEC were low under normoxic conditions, but were significantly increased after 5 h of hypoxia. Endothelial Rho-kinase expression was not detected until 3 h of hypoxia; such expression remained significantly increased after 5 h. On the other hand, endothelial eNOS expres-sion was similar after 3 h of hypoxia, but was significantly decreased after 5 h. The hypoxia-induced decrease in eNOS expression was significantly enhan-ced by expression of the constitutively active form of RhoA and significantly inhibited by suppression of RhoA expression by small interfering RNA. The hypoxia-induced decrease in eNOS expression was significantly inhibited when endogenous Rho-kinase activation was inhibited by Rho-binding domain expression. Conclusion: Expression and activation of RhoA and Rho-kinase inhibit eNOS expression in endothelial cells, attempts to down-regulate RhoA and Rho-kinase by multiple drugs, such as statins or Rho-kinase inhibitors, might provide endothelial and cardiovascular benefits through upregulation of eNOS.
