The Effect Of Ang-?-p22phox-ROS Pathway In The Pathogenesis Of Kawasaki Disease

Part 1 Study on the correlation among Ang-?,p22 phox andcoronary artery lesion in Kawasakai diseaseObjective:To explore the role of Ang-? and p22 phox in Kawasakai disease and coronary artery lesion.Methods:1.51 Kawasakai disease patients(15 patients with coronary artery lesions,36 patients without coronary artery lesions),28 healthy controls were enrolled into the study.The expression of plasma Ang-?level were measured using enzyme-linked immunosorbent assay(ELLSA);2.40 Kawasakai disease patients(15 patients with coronary artery lesions,25 patients without coronary artery lesions),20 healthy controls were enrolled into the study.The expression of p22 phox m RNA of the peripheral blood mononuclear cells(PBMCs)were measured using real-time polymerase chain reaction(RT-PCT).Results:1.There is no significant difference between the plasma concentrations of Ang-? in patients with coronary artery lesions and without coronary artery lesions(P>0.05),the plasma concentrations of Ang-? in patients with coronary artery lesions is higher than the normol controls(P<0.01),the plasma concentrations of Ang-? in patients without coronary artery lesions is higher than the normol controls(P<0.01).2.There is no significant difference between the expression of p22 phox m RNA of the peripheral blood mononuclear cells(PBMCs)in patients with coronary artery lesions and without coronary artery lesions(P>0.05),the expression of p22 phox m RNA of the peripheral blood mononuclear cells(PBMCs)in patients with coronary artery lesions is higher than the normol controls(P<0.05),the expression of p22 phox m RNA of the peripheral blood mononuclear cells(PBMCs)in patients without coronary artery lesions is higher than the normol controls(P<0.05).3.The expression of p22 phox m RNA of the PBMCs has a positve correlation with the plasma concentrations of Ang-?in KD(r=0.358,p=0.023).Conclusion:Ang-?,p22 phox may be involved in the pathogenesis Kawasaki disease and wether pariticipate in the development of coronary artery lesion in KD need a futher study.Part 2 Study of the effect of IVIG treatment on Ang-?and p22phox in Kawasakai diseaseObjective:To explore the mechanism of intravenous immunoglobulin(IVIG)in the treatment of Kawasakai Disease and the effect of IVIG on Ang-?,p22 phox in Kawasakai disease.Methods:1.51 Kawasakai disease patients who received IVIG treatment wereselected in the study,28 age-matched healthy children were selected as the control group.The expression of plasma Ang-?level were measured using enzyme-linked immunosorbent assay(ELLSA).2.40 Kawasakai disease patients(15 patients with coronary artery lesions,25 patients without coronary artery lesions)who received IVIG treatment were selected in the study,20 age-matched healthy children were selected as the control group.The expression of p22 phox m RNA of the peripheral blood mononuclear cells(PBMCs)were measured using real-time polymerase chain reaction(RT-PCT).Results:1.The plasma concentrations of Ang-? in pre-treatment is higher than post-treatment and control group(P<0.01).The plasma concentrations of Ang-? in post-treatment is higher than control group(p<0.01).2.The expression of p22 phox m RNA of PBMCs before IVIG treament is higer than after IVIG treament(P<0.05)and contol group(P<0.01).There is no significant difference between the expression of p22 phox m RNA of the peripheral blood mononuclear cells(PBMCs)in patients after IVIG treament and contol group(P>0.05).Conclusion:IVIG treatment affects both the plasma concentrations of Ang-?and the expression of p22 phox m RNA in acute stage.Part 3 In vitro injury effects of sera of patients with kawasaki diease on vascular endothelial cellsObjective:To investigate the in vitro effects of sera of patients with kawasaki diease on vascular endothelial cells,and explore their clinical significance.Methods:Human umbilical vascular endothelial cells(HUVEC)were treated with sera of patients with kawasaki diease(kawasaki diease serum group,n=36)and healthy controls(normal serum group,n=19)(volume fraction of 20%for each group)for 24 h,and blank control group was established.The cell morphology was observed by Inverted phase contrast microscope,cell apoptotic rates were determined by flow cytometry.The culture supernatant fluid of 10 patients was collected,the mass concentrations of Ang-? were measured by ELISA.and Real-Time PCR was employed to detect the expression of p22 phox m RNA in cells.Results:1.The number of HUVEC in pre-treatment and post-treatment kawasaki diease serum groups was smaller than those of blank control group and normal serum group,and condensed or fragmented nuclei and condensed cytoplasm were observed in kawasaki diease serum group.2.The cell apoptotic rate in pre-treatment kawasaki diease serum group is(18.06±2.35%),in post-treatment kawasaki diease serum group is(7.89±2.1%),in blank control group is(2.9±0.38%)and in normal serum group is(6.11±0.53%).3.There is no significant difference between the plasma concentrations of Ang-? in the mass concentrations of pre-treatment and post-treatmentkawasaki diease.4.The expression of p22 phox m RNA of HUVEC in pre-treatment serum group is higer than post-treatment and contol group(P<0.01).There is no significant difference between the expression of p22 phox m RNA of HUVEC in post-treatment and contol group(P>0.05).Conclusion:Treatment with sera of patients with kawasaki diease may change the morphology,inhibit the proliferation,promote the apoptosis of HUVEC,and the Increased express of the p22 phox m RNA may lead to the dysfunction of vascular endothelial cells.