Effect Of Fasudil On Hydrogen Peroxide-Induced TGFβ2 And IL8 After Endothelial Injury

Background: The current research indicates that the corresponding treatment of abnormal inflammatory mediators caused by oxidative stress is currently a research hotspot in the prevention and treatment of coronary heart disease.Fasudil has been proved to reduce endothelial cell injury by inhibiting the synthesis of inflammatory mediators IL-8 and TGF beta 2.RhoA-ROCK signaling pathway plays an important role in oxidative stress damage,so Fasudil may inhibit the synthesis of IL-8 and TGF beta 2 by inhibiting the RhoA-ROCK signaling pathway.Objective: To investigate the effect of fasudil(FH)on the expression of transforming growth factor-β2(TGFβ2)and interleukin-8(IL-8)in Hy926 human umbilical vein endothelial cells injured by hydrogen peroxide.Method: The cells were induced in cells incubated for 2 hours with a concentration of 30% hydrogen peroxide to make the oxidative damage model.Cells fused to 80-90% were passaged 1: 4 until the passaged cells fused to about 70%,which were used as experimental grouping cells.The cells were divided into control group(untreated normal cells),oxidative injury group(cells which hydrogen peroxide injuried for 2 hours and then replaced by 10% fetal bovine serum in DEME medium for 48 hours at 37 ℃ in a 5% CO 2 incubator),FH intervention group(wash oxidative damaged cells with PBS 2 times,then replaced with 75 nmol / L fasudil + medium at 37 ℃ for 48 hours in 5% CO 2 incubator),and FH alone group(uninjured cells with 75nmol/ L fasudil + medium,incubate the cells in a 5% CO 2 incubator at 37℃ for 48hours).The level of ROS was measured by fluorimetric method.The level of IL-8 and TGFβ2 was measured by ELISA.The assay of RhoA,MYPT-1p was measured by Western blot.Results: After the oxidative damage,the cells were apoptotic,and the cells became round and smaller.The number of apoptotic cells with the addition of fasudil was significantly lower than that in the oxidative damage group,but there was still apoptosis.The concentrations of ROS,IL-8 and TGF-β2 in oxidative injury group were significantly higher than those in control group,and the transcriptional levels of RhoA and MYPT-1p were significantly increased.Compared with the control group,the concentrations of ROS,IL-8 and TGF-β2 in the fasudil intervention group were significantly increased,and the transcription levels of RhoA and MYPT-1p significantly increased.Compared with the control group,the concentrations of ROS,IL-8 and TGF-β2,and the transcriptional levels of RhoA and MYPT-1p in the fasudil group alone has no significantly difference.However,compared with the oxidative injury group,the levels of ROS,IL--β2 concentration decreased,RhoA,MYPT-1p transcription level also decreased.Conclusion: Our data suggest that fasudil can effectively inhibit the inflammatory reaction caused by oxidative damage and inhibit apoptosis and it may results from the inhibition of the secretion of inflammatory cytokines after oxidative stress through RhoA-ROCK signaling pathway.