Effects Of Uric Acid On The Production Of ENos,ROS And Inflammatory Cytokines

Objective:In recent years,with the improvement of material life level and the change of people's lifestyle in our country,the incidence of hyperuricemia was increased.Hyperuricemia is caused by purine metabolic disorder,it is a metabolic disease.Hyperuricemia is not only the etiology of gout,but also closely related to the incidence of chronic kidney disease(CKD),cardiovascular disease,hypertension,and metabolic syndrome.Ross put forward the " response to injury hypothesis ",they suggest that various cytokine synthesis changes,inflammation activity,increased vascular permeability,a disturbed balance between vasodilator and vasoconstrictor activity.The endothelial dysfunction in hypertension,cardiovascular disease and chronic kidney disease has played an important role,however the relationship between hyperuricemia and endothelial cel s and its specific pathogenesis is not yet clear.To study the effects of different concentrations of uric acid on glomerular endothelial cel s and its possible mechanism,which provides a theoretical basis for clinical monitoring,early prevention and treatment of hyperuricemia.Methods:1.Different concentrations of uric acid stimulated HRGEC for 24 hours,the cell counting kit was used to test the activity of cel s.2.Different concentrations of uric acid(0,4,8,16 mg/dl)stimulated HRGEC with the time of 24 hours,the expression of eNOS?NF-?B p65?MCP-1?ICAM-1 were detected by Real-timePC R and Western blotting.Utilized the immunofluorescence methods to detect the expression of eNOS and NF-?B p65.3.Using the fluorescent probe DCFH-DA to detect the intracellular reactive oxygen species(reactive oxygen,species,ROS)after different concentrations of uric acid stimulated HRGEC for 24 hours.Results:1.Compared with the control group,with the stimulated of 8 mg/dl and 16 mg/dl of uric acid,the expression of mRNA and protein of eNOS was significantly down-regulated(P<0.05),there was no significant difference in the expression of eNOS mRNA and protein in 4 mg/dl of uric acid.2.The effect of uric acid(UA)on intracellular total reactive oxygen species(ROS)generation in HRGEC,compared with the control,the ROS levels were signifiantly increased after incubation with 8 mg/dl and 16 mg/dl of uric acid(P<0.05),but there was no significant difference in 4 mg/dl of uric acid.Compare with the 4 mg/dl of uric acid,the ROS levels were signifiantly increased after incubation with 8 mg/dl and 16 mg/dl of uric acid(P<0.05).3.HRGEC incubated with 8mg/dl and 16 mg/dl UA for 24 hours contained higher levels of NF-?B p65(P<0.05),however,there was no significant difference in the 4 mg/dl UA.Compare with the 4 mg/dl UA,the expression o f NF-?B p65 was significantly increased in the 8 mg/dl and 16 mg/dl UA(P<0.05).4.Compare with the control,the expression of MCP-1 ? ICAM-1 were significantly increased in the 8 mg/dl UA and 4 mg/dl UA(P>0.05),there was no significant difference in the 4 mg/dl UA.Compare with the 4 mg/dl UA,the expression of MCP-1?ICAM-1 was significantly increased in the 8 mg/dl and 16 mg/dl UA(P<0.05).Conclusions:1.High concentrations of uric acid may activate oxidative stress leads to glomerular endothelial injury,causing renal damage.2.High concentrations of uric acid may injury the endothelial cells through activateing the NF-?B pathway.