Effection Of Sulodexide On Podocin, CD2AP, Nephrin Expression In Kidney Of Membranons Nephropathy Rats

Objective: Membranous nephropathy (membranons nephropathy, MN)is an adult common cause of nephrotic syndrome, which pathology ischaracterized by limited glomerular subepithelial immune complex deposition.Clinical manifestations of massive proteinuria.This research buildsmembranous nephropathy model of SD rats by injecting Cationic BovineSerum Albumin(C-BSA),and applies Sulodexide to intervene, though theobservation of24-hour urinary protein of rats,and the expression ofmembranous nephropathy glomerular podocin, CD2AP, nephrin,discussingthe new target of Sulodexide on MN，in order to provide a new theoreticalbasis for the prevention and control of human MN。Methods:40eight-week-old healthy male SD rats, weight about160±20g,were fed adaptively for a week.After a week, the24-hour urine protein testwere normal.40rats were randomly divided into four groups of10, espectively,normal control group, model group, high dose Sulodexide group and low doseSulodexide group. In addition to the normal control group,after1week of thepre-immunization the other groups once every other day injected C-BSA viacaudal vein to copy MN rats model,normal control group injected with thesame dose of saline.The24-hour urinary protein were tested after continuousinjection for4weeks.After confirming the success of modeling,the high doseSulodexide group were given Sulodexide20mg/kg/d, low-dose group weregiven Sulodexide10mg/kg/d. Model group and normal control group giventhe same dose of saline, the rats free access to food, drinking water.All of thefour groups continued dosing for4weeks.After that,testing the24-hours urineprotein, then the rats were sacrificed. Renal tissues of rats were harvested forinvestigate ultrastruction changes by transmission electron microscopy andtesting the expression of nephrin， CD2AP and podocin by immunohistochemistry.Result1. General condition of experimental ratsThe rats of normal control group both diet and urine volume are normal,prompt response, bright eyes, and plump muscle.Afer injecting C-BSA viacaudal vein, the rest of the three groups rats all showed anorexia, lassitude,baldness and less activities.Some rats appeared scrotal edema.In the last weekof experiment, comparing with normal control group, the rats of the otherthree groups were smaller, and baldness (abdomen as the most important).2.24-hours urine proteinAfter4weeks, injection of C-BSA,the24-hour urine protein of the modelgroup, high dose Sulodexide group and low dose Sulodexide group were risedobvious than these of control group (P both <0.05). Administration after4weeks, the high-dose Sulodexide group and low-dose Sulodexide group24-hour urine protein were reduced, compared with the model group werestatistically significant (P both <0.05). Compared with low-dose Sulodexidegroup, the24-hour urinary protein of high dose Sulodexide group weredeclined more significantly (P <0.05).3. Pathomorphism of renal tissuesLight microscope: The control group had no obvious abnormalities, hadno cells increase, the capillary loop open finely.The basement membrane hadlittle thickness. The model group showed that the glomerular basementmembrane had thickened evidently, the cells increased. Comparing with themodel group, the thickened basement membrane in low dose Sulodexidegroup and high dose Sulodexide group relatively relieved, the cells of thesetwo groups increased mildly.Electron microscope: The rats of normal control group had a clearstructure of podocyte and aligned foot processes.The model group showedbasement membrane thickened significantly, similar electron dense materialdeposited under the glomerular epithelium, foot processes fused diffusely orlost. The glomerular basement membrane of the low-dose Sulodexide group had thickened irregularly, similar electron dense material deposited under theglomerular epithelium, part of podocyte foot processes began to fuse. Thehigh-dose Sulodexide group showed that the glomerular basement membranehad thickened mildly, glomerular subepithelial electron-dense material were alittle. Part of podocyte foot processes began to fuse.4. The results of immunohistochemical stainingImmunohistochemisty staining show that the expression of podocin，CD2AP and nephrin located under the glomerular epithelium. Podocin，CD2AP and nephrin of normal control group were normal. These of the otherthree groups were reduced in diffient degrees,compared with the normalcontrol group， statistical significance(P both<0.05). The expression ofpodocin，CD2AP and nephrin were down-regulated in the model groupcompared with low-dose Sulodexide group and high-dose Sulodexide group,statistical significance(P both<0.05). Compared with high dose Sulodexidegroup, the expression of podocin，CD2AP and nephrin in the low doseSulodexide group were down-regulated,statistical significance (P both<0.05).Conclusion:1.The expression of podocin，CD2AP and nephrin in the model groupwere signiflcantly down-reguIated and correlated to podocytes lesion andurine protein in MN rats. Podocin，CD2AP and nephrin may play a role inpodocyte processes fusion and developing urinary protein excretion in MNrats.2. Sulodexide can decline urine protein of MN rats.3. Sulodexide can increase the expression of podocin，CD2AP andnephrin on podocytes of membranous nephropathy rats. The therapeutic actionis probably relative to maintaining the integrity of podocyte slit membranestructure and glomerular filtration function.4. Sulodexide therapy urinary protein of MN rats and protect kidney had dose dependent.