Relation Of Expression MMPs,TIMPs And NO With Coronary Artery Lesions In Kawasaki Disease

Objective:To explore the expression of MMP-2,MMP-3,TIMP-1, TIMP-2 and NO in acute phase and convalescence in Kawasaki Disease and their relationship during the development of KD.And discuss their expression in the coronary artery lesion (CAL) with KD,thus to provide some theory evidence for prevention the CAL.Method:1. Serum collection (1) Work group:The serum was collected from 30 KD children(15 patients with coronary arterial lesions,group CAL;and 15 patients with no coronary arterial lesions,group NCAL.) with KD in the acute phase (pathogenesis 1-2 weeks,before IVIG) and convalescence phase (pathogenesis 3-4 weeks, after IVIG). (2) Control group:30 children with febrility (group F) and 30 normal children (group C)2. Detect index:The levels of MMP-2,MMP-3,TIMP-1,TIMP-2 were detected by Enzyme-linked immunosorbent assay(ELISA) in serum.The level of NO was measured by nitrate reductase spectrophotometry in serum.3. Statistical analysis:Statistical anaylsis was peformed using the statistical programs SPSS 17.0, data were expressed as average±standa- rd devidation. Variance with t test, neterogeneity of variance with t' test. The ratio of MMP-2/TIMP-2,MMP-3/TIMP-1 with rank test. The correlation analysis with MMPs,TIMPs and NO to adopt Pearson.Size of test is 0.05 (bilateral).P values less than 0.05 is considered to be statistical significance.Result:1. MMP-2,TIMP-2,MMP-2/TIMP-2: (1) MMP-2 of group CAL is (480.75±9.82μg/L),compared with group NCAL (450.06±4.38μg/L),is significantly higher than that in group NCAL (P<0.05); TIMP-2 of group CAL is (393.61±3.89μg/L), MMP-2/TIMP-2 of group CAL is (1.57±0.21), compared with group NCAL TIMP-2 is (376.30±11.28μg/L), MMP-2/TIMP-2 is (1.29±0.14), are significantly higher than that in group NCAL (P < 0.01). (2) Before IVIG therapy serum MMP-2 level is (465.4±17.31μg/L), TIMP-2 level is (384.95±12.09μg/L) and the ratio of MMP-2/TIMP-2 is (1.71±0.16) are significantly elevated in KD than that in group F and group C (P<0.01). (3) After IVIG therapy serum MMP-2 level is (422.51±16.13μg/L),TIMP-2 level is (338.83±14.93μg/L),MMP-2 level in group F is (311.26±15.81ug/L), TIMP-2 level is (264.05±12.35μg/L), and MMP-2 level in group C is (295.02±18.18μg/L), TIMP-2 level is (253.46±15.41μg/L), After IVIG therapy serum MMP-2 and TIMP-2 level are significantly higher than group F and group C (P<0.01).After IVIG therapy serum MMP-2/TIMP-2 level is (1.29±0.12), compared with group F (1.05±0.19) and group C (0.94±0.12) statistical significance was accepted for p values of<0.05. (4) Before IVIG therapy serum MMP-2 level is (465.40±17.31μg/L), compared with after IVIG therapy serum MMP-2 level is (422.51±16.13μg/L), is significantly higher than after IVIG therapy(P<0.05). Before IVIG therapy serum TIMP-2 level is (384.95±12.09μg/L), the ratio of MMP-2/TIMP-2 is (1.71±0.16), compared with after IVIG therapy serum TIMP-2 is (338.83±14.93μg/L),MMP-2/TIMP-2 is (1.29±0.12), statistical significance was accepted for p values of<0.01.2. MMP-3,TIMP-1,MMP-3/TIMP-1: (1) MMP-3 of group CAL is (58.38±9.43μg/L) compared with group NCAL (33.78±3.34μg/L),is significantly higher than in group NCAL (P<0.01).TIMP-1 of group NCAL is (357.16±22.79μg/L), compared with group CAL (298.37±61.45μg/L) is significantly higher than in group CAL (P<0.01); MMP-3/TIMP-1 of group CAL is (0.45±0.15),compared with group NCAL(0.08±0.02), is significantly higher than in group NCAL (P<0.05). (2) Before IVIG therapy serum MMP-3 level is (46.07±14.31μg/L), the ratio of MMP-3/TIMP-1 is (0.14±0.06), compared with after IVIG therapy serum MMP-3 level is(32.41±7.48μg/L), the ratio of MMP-3/TIMP-1 is(0.13±0.09),are significantly elevated in KD than that in after IVIG therapy, group F and group C (P<0.01). (3) Before IVIG therapy serum TIMP-1 level is (328.93±45.17μg/L), after IVIG therapy serum TIMP-1 level is(281.61±39.42μg/L),compared with group F(247.64±16.73μg/L) and group C (94.63±3.60μg/L), are significantly higher in KD than that in group F and group C (P<0.01). (4) Group F serum MMP-3 is (31.96±4.11μg/L),TIMP-1 is (247.64±16.73μg/L) compared with group C, MMP-3 is (10.51±0.45μg/L), TIMP-1 (94.63±3.60 u g/L), are significantly higher than that in group C (P<0.01).The ratio of MMP-3/TIMP-1 in group F is (0.15±0.04), compared with group C (0.12±0.02),there is no statistical significance (P>0.05)3. NO: (1) NO of group CAL is (176±22μmol/L),compared with group NCAL (97±14μmol/L),is significantly higher than that in group NCAL (P<0.05). (2) Before IVIG therapy serum NO level is (137±44μmol/L),after IVIG therapy serum NO level is (73±23μmol/L), compared with group F (44±11μmol/L),group C is (41±11μmol/L),are significantly higher than that after IVIG therapy,group F and group C(P<0.01).(3)Before IVIG therapy serum NO level is (137±44μmol/L),compared with after IVIG therapy (73±23μmol/L),is significantly higher than that in after IVIG therapy (P< 0.01). (4) NO of group F is (44±11μmol/L), compared with group C (41±11μmol/L), there is no statistical significance (P >0.05)4. Associativity analysis with MMPs,TIMPs and NO:(1) There were significant positive correlations between NO and MMP-2,MMP-3 levels in KD patients. (r=0.895,P<0.01; r=0.898,P<0.01). (2) There were significant negative correlations between NO and TIMP-2, TIMP-1 levels in KD patients. (r=-0.382,P<0.05; r=-0.772,P< 0.01)Conclusion:1.MMP-2,TIMP-2,MMP-2/TIMP-2 in serum in group CAL were significantly higher than that in group NCAL,suggesting that MMP-2,TIMP-2 in serum and the ratio of MMP-2/TIMP-2 might be risk factors of KD coronary artery lesion.These findings indicate that they may play an important role in KD with vessle wall damage and CAL.There should be MMP-2,TIMP-2 to heighten after acute stage in KD,suggesting that it may be exist an agelong coronary artery injury. So we need to monitoring blood vessle damage in KD patients.2.MMP-3 of group CAL is significiantly higher than group NCAL, suggesting that MMP-3 probably to participate in the damage process of vessle wall.Therefore,the measurement of serum MMP-3 might be of important clinical value in the prediction and the early diagnosis of KD with coronary artery lesion.In acute stage serum MMP-3 and TIMP-1 levels and the ration of MMP-3/TIMP-1 were higher in patients with KD than convalescent stage and control group; suggesting that these data are important objective markers with diagnosis KD in early stage.3.The overexpression of NO may play an important role in the development of vasculitis in acute stage of KD, above all coronary artery lesion;suggesting that NO level in serum should be an important objective markers of KD coronary arterial lesion.4. There were significant positive correlations between NO and MMP-2.MMP-3 levels in KD patients,suggesting that the interaction between MMP-2,MMP-3 and NO,to promote the development with CAL in KD.There were significant negative correlations between NO and TIMP-2,TIMP-1 levels in KD patients,suggesting that TIMPs work as inhibitor of MMPs,to inhibition the MMPs in certain degree,they inhibition the expression of NO invariably at the same time. We could regard as a new research direction to diagnosis and treatment KD with CAL.To provide the theory accordance for prevent the CAL.To make use of TIMPs regulation the level of MMPs, meanwhile,it may be regulate the level of NO in serum.