Effects Of Sulodexide On Expression Of Podocalyxin,podocin, WT1in Podocytes Of Adriamycin Nephropathy Rats

Nephrotic syndrome (nephritic syndrome, NS), is a group of clinicalmanifestation caused by serious proteinuria which due to many kidneypathological damage.Some patients even can progress to end-stage renaldisease,that bring heavy burden to the family and society. According to thepathogenesis, NS is divided into primary and secondary.The changes ofglomerular filtration membrane permeability including charge barrier andpore diameter barriers, especially the podocyte proteins in the diaphragm areamolecular abnormalities are related to the primary NS. Neighboring footprocesses are connected by a specialized cell-cell junction, the glomerular slitdiaphragm, which represents the main size-selective filter barrier in thekidney. The slit diaphragm is thought to be a modified adherens junction. Theslit diaphragm is known to include nephrin,podocin,NEPH1,ZO-1,CD2AP,FAT,P-cadherin, Podocalyxin carries a negative charge which rejectseach other and can prevent the neighboring foot processes fromadhesioning.Podocin is a one of the membrane protein specifically expressedin glomerular podocytes and its function displays as the ion channels andsignal transduction. WT1anchor the slit diaphragm to the podocyte.Recentstudies found that, many of glomerular disease are related WT1.The most important treatment of NS is to reduce the proteinuria.Corticosteroids and immunosuppressive agents are the mainly treaments atpresenst. Although the corticosteroids curative effect is certainly, its sideeffects are also obvious and the patients are tending to non-adherence. So thehot study is to finding out new drugs that can slow the side effects while assistthe treament.Sulodexide, is a glycosaminoglycan with anticoagulant andantithrombotic activities. According to the reports, GAGs can maintain thenegative charge on the capillary wall, repair endothelial injury and the defects of matrix metabolism,promote the synthesis of basemen, restore itspermeability and then reduce the urine protein feom leaking out.Objective:1To observe the expression of podocyte membranecomponent podocalyxin,podocin,WT1in rats with adriamycin nephropathy(AN);2To study the curative effects of sulodexide on AN and its effects on theexpression of podocyte podocalyxi,podocin,WT1.Methods:1Rat models of AN were established by a single tailintravenous injection of adriamycin (6.5mg/kg), The AN model wasconsidered to be successful when24h urinary protein were significantlyincreased.260healthy male SD rats were randomly divided into five groups:control group(group A), AN group(group B), prednisone treatmentgroup(group C)with prednisone7.5mg/kg/d, sulodexide treatmentgroup(group D) with sulodexide(10mg/kg/d)and prednisone(7.5mg/kg/d)combined with sulodexide(10mg/kg/d)treatment group(group E). At the end of4,8week,collecting the24h urine, blood and renal tissue samples, testingurinary total protein(UTP), serum albumin,cholesterol,triglyceride,creatinineand blood nitrogen. The ultrastruction changes were determined by bytransmission electron microscopy. The expression of podocalyxin,podocin,WT1in glomerular podocyte were detected by immunohistochemistry andRT-PCR. The results were analyzed with an image-processing system.All thedata were analyzed by SPSS13.0statistics software, P value<0.05wasconsidered to have statistical significance.Results:1Compared with control group, AN rat model was madesuccessfully presenting with edema,poor appetite,higher UTP(P＜0.05).Ultrastructure changes in AN rats showed that podocyte processeseffacement,GBM nonuniform,endothelial cells tumidness at the end of4and8week.2Compared to group B, prednisone(group C), sulodexide(group D) andprednisone combined with sulodexide(group E) could decrease UTP, the level of serum cholesterol,triglyceride and increase the serum albumin level(P＜0.05). Treatment with sulodexide combined with prednisone got even lowerUTP and higer serum albumin level compared to group C and D.3At the end of the8th week, compared to group A, ultrastructure studyshowed that there were significant changes in AN groups, includingpodocyte processes effacement,GBM nonuniform,endothelial cells tumidness.Compared with group B, group C, D, E had less severe lesion while group Ehad even mild lesion.4Immunohistochemistry and RT-PCR studies showed that the expressionof podocalyxin,podocin,WT1at the end of the4th and8thweeks wassignificant decreased in AN ratscompared with the normal glomeruli whiletheir expression were obviously increased in group C,D,E than groupB(p<0.05)). The additive effects were obtained when sulodexide andprednisone were combined.Conclusion:1The expression of podocalyxin,podocin,WT1in renaltissue in AN rats were downregulation,corelates with UTP and renaldamage,suggesting that podocalyxin,podocin,WT1may correlated with NS.2Sulodexide can improve NS symtom and ultrastruture lesion maybethrough restoring the expression of podocalyxin,podocin,WT1to maintainthe integrity of podocytes.