Effect of fasudil on the human umbilical vein endothelial cells injury induced by lipopolysaccharide

Objective: To observe the effects of fasudil on the human umbilical vein endothelial cells(HUVECs) injury induced by lipopolysaccharide(LPS),explore its mechanism of action.Methods: HUVECs were obtained from human umbilical vein,digested with 0.1% type Ⅰ collagenase,and cultivated and passaged in endothelial cell culture medium.The cell morphology was observed under the inverted microscope,and the cells were identified using immunohistochemistry.HUVECs were randomly divided into the control group(cultured with endothelial cell medium),LPS group(cultured with 0.1 μg/mL LPS) and fasudil group(cultured with 3.275 μg/mL fasudil and 0.1 μg/mL LPS).The protein and mRNA levels of RhoA,ROCK2 and p-ERM were detected using Western blot and quantitative real-time PCR,respectively.Results: The primary cells could adhere to the plate after 24 hours of inoculation,and arranged as paving stone on the third day.The HUVECs identified using the immunohistochemistry staining of Ⅷ factor.related antigen.The protein and mRNA levels of RhoA,ROCK2 and p-ERM in LPS group were significantly higher than those in control group(P<0.01).The protein and mRNA levels of ROCK2 and p-ERM in fasudil group were lower than those in LPS group(P<0.01),the differences of the protein and mRNA levels of RhoA between fasudil group and LPS group were not statistically significant(P>0.05).Conclusions: The Rho/ROCK/ERM signaling pathway plays an important role in the cytoskeletal damage in HUVECs induced by LPS.Fasudil can inhibit the LPS-induced cytoskeletal rearrangement injury,the mechanism of which may be related to inhibiting the Rho/ROCK signaling pathway activation.