The Expressions Of MMP-2,9 And TIMP-1 In Rats Vascular Calcification With Chronic Renal Failure

Part One The Expressions of MMP-2,9 and TIMP-1 in Rats Vascular Calcification with Chronic Renal FailureObjective: To establish vascular calcification model of rat with Chronic Renal Failure; and to examine whether MM-P2,9 and TIMP-1 is involved in vascular calcification in chronic renal failure. Methods:(1)Chronic renal failure was induced in Wistar rats by received stomach irritation daily with 2% adenine 250-300mg/kg for 6 weeks.(2)Applying the methods of RT-PCR ,we observed the changes of MMP-2,9 and TIMP-1 mRNA .(3)Von Kossa stained vessels for the presence of calcification.( 4) TIMP-1,OPN,Type I andâ…£Collagen were analyzed by immunhistochemistry. All above and serum biochemical parameters were examined at 2,4,6 and 8 weeks.Results:(1)Serum urea nitrogen,creatinine concentrations and inorganic phosphate progressive increased significantly (P<0.05) in all model rats compared with those of control rats after 2 weeks,and calcium-phosphorus product was increased also.(2) Medial calcification was found in the aorta,femoral,renal artery and coronary arteries after 6 weeks.(3)MMP-2/TIMP-1 ratio was lower than control group,and MMP9/ TIMP1 ratio showed opposite changes.(4)The expression of TIMP-1 in calcified aortic smooth muscle cells in the medial layer was increased obviously by immunhistochemistry, and there were positive immunostaining for OPN and type I collagen increased,but absent staining for type IV collagen in these same areas .Conclusion : The morphology of arteriosteogenesis in uraemic rats made by adenine approximate to uraemic patient, so it may serve as a useful model for arteriosteogenesis of chronic renal failure . The imbalance of MMP-2,9 /TIMP-1 involved in vascular calcification of chronic renal failure.PartTwo Relationship between serum,urine tissue inhibitor of metalloproteinases-1(TIMP-1) levels and the stages of CKDObjective: To investigate the relationship and significances of serum and urine TIMP-1 level and different stages of CKD.Methods: Serum and urine level of TIMP-1 were measured by enzyme-linked immunoadsorbent assay ( ELISA ) double-antibody- sandwich method in 122 CKD patients and 40 healthy controls,CystC,PTH and LDL were determined simultaneously . Correlationanalysis between TIMP-1 level and degree of renal function were performed.Results: Serum and urine TIMP-1 in CKD group was significantly higher than that healthy control group(p<0.01). There was correlation between serum,urine TIMP-1 and different stages of CKD. Especially,TIMP-1 in serum obviously increased in early stage of CKD.The corelation of Serum and urine TIMP-1 with glomerular filtration rate was positive( r=-0.647,-0.437,respectively, p<0.01).Conclusion : Serum concentrations of TIMP-1 may be used as a fine potential maker for diagnosis of renal fibrosis.