Study On The Role And Mechanism Of Uric Acid-induced HCAEC Injury

Objective: In this study,human coronary artery endothelial cells were cultured with high,medium and low concentrations of uric acid to investigate whether different concentrations of uric acid could induce endothelial cell damage and its mechanism.Method:(1)In order to explore the effect of different concentrations of uric acid on HCAEC,logarithmic growth phase HCAEC was taken and divided into normal control group,PBS group,2.5mg / dl UA group,5mg / dl UA group,10 mg / dl UA group,20 mg / dl UA group.(1)MTT method was used to detect cell survival rate;(2)RT-PCR was used to detect the m RNA expression level of intercellular adhesion molecule ICAM-1;(3)Western blot was used to detect ICAM1 protein expression level;(4)ELISA method was used to detect the protein expression of inflammatory factors IL-1 and hs-CRP Level.(5)Fluorescent probe DCFH-DA was used to detect the expression of reactive oxygen species.(2)To explore the mechanism of endothelial cell injury induced by hyperuric acid,(1)Take HCAEC in logarithmic growth phase and divide it into normal control group,20 mg / dl UA group,PDTC + 20 mg / dl UA group,and detect ICAM-1 m RNA expression level and Protein expression levels,IL-1,hs-CRP protein expression levels.(2)Take HCAEC in logarithmic growth phase and divide it into normal control group,20 mg / dl UA group,NAC + 20 mg / dl UA group,and detect the expression of reactive oxygen species with fluorescent probe DCFH-DA.Results:(1)With the increasing concentration of UA,the cell survival rate is gradually increased;high concentration of uric acid can induce the RNA level of adhesion molecule ICAM-1 and the protein expression of protein,inflammatory factor hs-CRP and IL-1?(P<0.05),low concentration and medium concentration of uric acid could not induce the RNA level of adhesion molecule ICAM-1 and the protein expression of protein,inflammatory factors hs-CRP and IL-1?.High concentration of uric acid could induce the expression ofreactive oxygen species ROS.High(P<0.05),low and medium concentrations of uric acid could not induce an increase in the expression of reactive oxygen species.(2)NF-?B blocker PDTC can inhibit the increase of inflammatory factors caused by high uric acid(P<0.05);antioxidant NAC can inhibit the increase of reactive oxygen species caused by high uric acid(P<0.05).Conclusion: High uric acid can cause HCAEC damage by inducing inflammatory factors and increased expression of reactive oxygen species.Inhibition of the activation of the NF-?B pathway produces anti-inflammatory effects,and antioxidants can inhibit oxidative stress caused by high uric acid.