| Objective: In this study,the obstructive Nephropathy rat model was established by unilateral ureteral obstruction(UUO)in which the Rennin-angiotensin-aldosterone system(RAS)and the aldosterone wereactivated,leading to the inflammatory injury and interstitialfibrosis of the kidney.By means of observing the express of the Serum and Glucocorticoid-induced Protein Kinase 1(SGK-1),and detecting thevariation of the index of correlationas Monocyte Chemotactic Protein 1(MCP-1),F4/80,Transforming Growth Factor-?1(TGF-?1),to study the role of eplerenone in resistance to inflammatory injury and interstitialfibrosis and the mechanism of the Shenluotong in resistance tothe interstitialfibrosis of the kidney.Methods: Forty-eight healthy female Wistar ratsweightingabout 200±20g wereused in this study.The experimental rats ate and drank freely and temperature was controlled in(22±2)?.After one weekadaptive feeding,rats were randomly divided into sham group,UUO group,EPL group,and SLT group,for 12 rats in each group.Then,all rats were copied with the unilateral ureteral obstructionnephropathy modelexcept for the sham group:after anesthesia with 10% chloral hydrate,the rats were sliced through the left middle abdominal skin,separated the left ureter,ligated respectively on ureteral 1/3 and middle 1/3 ligation with silk,cut off the ureter.Then sutured in skin by layer to layer.The rats in Sham group were only separated the left ureterbut not ligated and cutted off the ureter.After operation,the rats were treated with 100mg/(kg.d)Eplerenone in fodder in EPL group and 26g/kg in SLT group;others in sham and UUO group administered with the same volume of saline once a day.Continuous intervention was for 10 days.Beheaded after weigh the rats 10 days later,retain blood specimens for detection.Cut down the obstruction kidney,weigh the tissue after remove integument,part of the kidneys were fixed by 4% paraformaldehyde and part of the kidney keep at-70 for extracting RNA and protein.The expression of ?SGK-1 ? MCP-1 ? TGF-?1 and F4/80 were detected by the way of immunohistochemistry and Western blot.Results:1 The general situation of the ratsThe general situation of Sham group's rats were in good condition:daily diet and water were stable,the growth of weight was gradual,hair was shiny,activity was agile,and response to external stimuli was sensitive.The water intake,food intake and weight were down-regulated in UUO group,the activity was also less and the hair was loose and reluster and the reaction to outside stimulus was lagged.10 days after the surgery,the weight of the model rats increased slowly,and exhibit significant difference to the rats in the sham group(P <0.01),the weight of the kidney of the model rats increased significantly and shows significant difference to the rats in the sham group(P<0.01);the weight of the Obstruction side kidney of the rats in eplerenone and Shenluotong group decreased significantly compared with the UUO group(P <0.05).2 The change ofPathomorphology in renal tissueHE staining showed that tubular epithelial cells arranged regularly and it had few inflammation cells in Sham group.There were a large number of inflammation cells infiltration,and renal tubular epithelial cells were denatured,shed and turgid in UUO group.The inflammation cell infiltration was significantly reduced and renal tubular epithelial cells were turgid in EPL and Shenluotong groups.Masson staining showed that a little collagen deposition in glomerular/tubular basement membrane and renal interstitium in Sham group and the Glomerular and renal tubular structure is clearThe structure were disordered and tubular interstitial collagen were significantly increased in UUO group.Compared with the sham group,there were some degree of collagen deposition in treatment groups,but it was significantly reduced compared with the UUO group.3 The expression of SGK-1?MCP-1?F4/80?TGF-?1In Sham group,the expression were weak of SGK-1 mainly in distal renal tubular epithelial cells;In UUO group,the expression was enhanced mainly in renal tubular epithelial cells and interstitium cells;In ELP and SLT groups,the expression were positive but significantly decreased in range and strength.In Sham group,the positive expression of MCP-1 was only in the cytoplasm of the renal tubular epithelial cells which showed weakly positive expression.In UUO group,the expression of MCP-1 were remarkably increased;Compared with the rats in Sham group,the expression of ?-SMA were enhanced in both EPL group and SLT group,but compared withUUO group was reduced obviously.In sham group,F4/80 was occasional expressed;the positive cells in UUO group significant increased mainly in renal tubular epithelial cells and interstitiumcells;Compared with UUO group,the positive cellsin EPL and SLT groups were significant decreased mainly in renal interstitium.In sham group,the TGF-?1 expressed weekly,mainly in renal tubular epithelial cells;In UUO group,the expression of TGF-?1 increased,mainly in the cytoplasm of the proximal convoluted tubuleepithelial cells;In EPL and SLT groups,the TGF-?1 showed moderate expression,but markedly reduced compared with UUO group.Using Western blot method to detect the expression of SGK-1?MCP-1?TGF-?1:in UUO group,the expression of SGK-1?MCP-1?TGF-?1 was significant enhanced compared with the sham group(P<0.01);in EPL and SLT groups,the expression was significant decreased(P<0.05 or P<0.01).Conclusion:1 obstructive nephropathy can lead to the inflammatory injury of the kidney by means of inducing the activation of aldosterone and then activating the expression of the downstream signal molecules SGK-1.The eplerenone and Shenluotong can down-regulate the expression of SGK-1,future reduce the secretion of MCP-1 and inhibit microphage infiltration,then delay kidney damage.2 Eplerenone and Shenluotong can inhibit macrophage infiltration,down-regulate the expression of TGF-?1,thus reducing the deposition of ECM,to delay the process of renal interstitial fibrosis. |
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